Biotechnology Advances

Biotechnology Advances 37 (2019) 319–339
Contents lists available at ScienceDirect
Biotechnology Advances
journal homepage: www.elsevier.com/locate/biotechadv
Research review paper
Newly isolated microorganisms with potential application in biotechnology T

a b a,c b
Marina G. Pessôa , Kele A.C. Vespermann , Bruno N. Paulino , Mayara C.S. Barcelos ,
⁎
Glaucia M. Pastorea, Gustavo Molinaa,b,
a
Laboratory of Bioflavors, Department of Food Science, School of Food Engineering, University of Campinas, Campinas, São Paulo, Brazil
b
Laboratory of Food Biotechnology, Food Engineering, Institute of Science and Technology, UFVJM, Diamantina, Minas Gerais, Brazil
c
Faculty of Pharmaceutical Sciences, Federal University of Amazonas, Manaus, Amazonas, Brazil
A R T I C LE I N FO A B S T R A C T
Keywords: Nowadays, food, cosmetic, environmental and pharmaceutical fields are searching for alternative processes to
Bioprospecting obtain their major products in a more sustainable way. This fact is related to the increasing demand from the
Novel microorganisms consumer market for natural products to substitute synthetic additives. Industrial biotechnology appears as a
Isolation techniques promising area for this purpose; however, the success of its application is highly dependent of the availability of
Screening studies
a suitable microorganism. To overcome this drawback, the isolation of microorganisms from diverse sources,
Aromas
Biosurfactants
including fermented food, adverse environments, contaminated samples or agro-industrial wastes is an im-
Polysaccharides portant approach that can provide a more adaptable strain able to be used as biocatalyst and that exhibit
Microbial lipids resistance to industrial conditions and high yields/productivities in biotechnological production of natural
Biotechnological products compounds. The aim of this review is to provide a solid set of information on the state of the art of isolation and
screening studies for obtaining novel biocatalysts able to produce natural compounds, focusing in aromas,
biosurfactants, polysaccharides and microbial oils.
1. Introduction applications with the development of a spectrophotometric screening

method for avermectin oxidizing microbes (Bertrand et al., 2014; Wang
The biotechnological production of ingredients emerges as an at- et al., 2017). It was also shown the development of a fast and simple
tractive alternative for industrial purposes, since it occurs at mild method for rapid screening of phosphate accumulating microorganisms
conditions, presents high regio- and enantio-selectivity and do not that allow a large number of strains to be screened based on color assay
generate toxic wastes (Pessôa et al., 2017). Additionally, the biopro- (Chaudhry and Nautiyal, 2011).
ducts obtained may be labeled as “natural” when obtained from mi- In some areas, mutations and metabolic engineering techniques are
croorganisms, plants, or animal cells at a relatively low cost being used, for example for selection of potential lactic acid producing
(Vespermann et al., 2017). Among these biocatalysts, microorganisms strains through the development of a high automated colorimetric
are of particular interest because of their great metabolic diversity for method based on sequential enzymatic reactions (Liaud et al., 2014).
modifying and upgrading a variety of complex organic molecules Several molecular methods to study the microbiota of soil and the
(Palmerín-Carreño et al., 2015). In whole cell biocatalysts, membranes mycosphere are being evaluated, such as metagenomics and meta-
and walls protect the enzymes from shear forces and other factors while transcriptomics combined with cultivation-based approaches (Van Elsas
cofactors can be regenerated within the cell under certain conditions and Boersma, 2011). On the other hand, these advanced techniques for
(De Carvalho and da Fonseca, 2006). screening can still be expensive for this purpose and complicated with a
In this sense, screening assays must be developed and need to be lack of suitable compounds for high-throughput assays, leading to new
established considering challenging substrates, process conditions and strategies such as the development of chromogenic probes for efficient
products diversity, quantity and purity (Emmerstorfer-Augustin et al., screening and evaluation of feruloyl esterase-like activities. The hy-
2016). Thus, different methods and strategies have been developed and drolysis of the chromogenic compounds substrates leads to a color
proposed in the technical literature in the most diverse areas: from the change which can be readily monitored in both qualitative solid
search of novel bioactive compounds for the field of drug discovery medium-based and quantitative liquid assays (Gherbovet et al., 2016).
using genome sequencing and microorganism co-culture, to agricultural In this way, the selection of new isolates is a wide field of
⁎
Corresponding author at: Laboratory of Food Biotechnology, Institute of Science and Technology - Office 334, UFVJM, Diamantina, Minas Gerais, Brazil.
E-mail address: gustavomolinagm@gmail.com (G. Molina).
https://doi.org/10.1016/j.biotechadv.2019.01.007
Received 7 April 2017; Received in revised form 15 January 2019; Accepted 15 January 2019
Available online 18 January 2019
0734-9750/ © 2019 Elsevier Inc. All rights reserved.
M.G. Pessôa et al. Biotechnology Advances 37 (2019) 319–339
Fig. 1. Natural compounds obtained through microbial biotechnology and its applications. Newly isolated microorganisms are a versatile source of value-added
products due to their metabolic characteristics and robustness generally related to their original environments. The four compounds highlighted in the present review
(aromas, biosurfactants, polysaccharides and microbial oils) have been extensively studied considering the broad applications in several industrial fields.
Fig. 2. Chemical structure of aroma compounds obtained in fermentation processes.
commercial exploration, besides opening several possibilities for in- Despite of the great diversity of methods that have been developed
dustrial bioprocesses development targeting for fine chemical produc- for the selection of new biocatalysts for the most diverse areas of re-
tion, that are increasingly relying on cell factories developed through search, in this material we analyzed the potential of the selection of
metabolic engineering and synthetic biology. The use of high microorganisms for the future of industrial biotechnology in the pro-
throughput techniques and automation for the design of cell factories, duction of important ingredients, such as biosurfactants, bioflavors,
and especially platform strains, has played an important role in the microbial lipids and polysaccharides (Fig. 1). In this sense, we provide
transition from laboratory research to industrial production, aiming to an overview of the current status of microbial selection describing
facilitate the process and the analysis in some types of application numerous processes that achieved promising results through long
(Jullesson et al., 2015). screening processes for a wide variety of industries including food,
320
chemical, pharmaceutical and cosmetic. We also present the conditions respectively; γ-decalactone using Sporobolomyces sp.; 1-octen-3-ol with
used during the screening trials, process conditions, strains found, and Neurospora sp. strain; 2-phenylethanol with Geotrichum fragrans; ben-
the main characteristics that make the development of these biopro- zaldehyde using Basidiomycetes sp.strains, vanillin by Escherichia coli,
cesses important for the future of the biotechnology industry. Schizosaccharomyces pombe and Saccharomyces cerevisiae (Dionísio et al.
2012).
2. Biotechnological potential of new selected microorganisms for In the biotransformation, a precursor is added in the process and the
the production of value-added compounds microorganism is induced to follow a specific pathway, reaching the
final product in one or two chemical reactions (Molina et al., 2012). In
2.1. Bioflavors addition, there is the possibility of using agro industrial byproducts as
nutrient and/or substrate sources in fermentation processes, lowering
Flavors and aromas are widely applied in food, cosmetic, chemical fermentation costs and making these processes viable industrially (Bicas
and pharmaceutical industries in order to supplement, enhance, or et al., 2009; Sarma et al., 2014).
modify the original taste/aroma of the product, and thus, presenting Among the substrates for biotransformation, terpenes can be high-
great importance over the acceptance of products by the consumer lighted since they are secondary metabolites of plants and can be found
market (Bicas et al., 2010; Palmerín-Carreño et al., 2015; Burdock, in byproducts of fruits/plants processing. Mono (limonene, α- and β-
2010). These compounds are remarkable for their volatility and che- pinenes) and sesquiterpenes (valencene and farnesene) are the most
mical diversity, including lactones, hydrocarbons, alcohols, ketones, common volatile compounds, constituents of a variety of essential oils.
vanillin, terpenes, aldehydes and esters (Molina et al., 2016; Fanaro Structurally, they are related to aroma compounds with great interest in
et al., 2016). Moreover, they differ in solubility, volatility and tem- industry, therefore, only few chemical reactions are necessary to obtain
perature and pH stability (Berger, 2015; Berger, 2007). a high-value product (Molina et al., 2014). Regarding economic aspects,
It has been reported at least 8000 identified volatile compounds the advantages of the biotransformation are clear when comparing the
comprising all these groups (some of them are shown in Fig. 2); reference prices of substrates and products, for example, R-Limonene
therefore, there is a great variety of possible combinations and in- has a reference price of US$ 34/L, while the price of its oxygenated
tensities (aroma perception) in formulations, resulting in a unique form, carveol, is around US$ 530/L (Molbase database, 2015).
aroma description for each product. Due to its low detection thresholds Another important step in bioprocess for natural aroma production
(generally in parts per billion–ppb), aroma compounds are usually is to choose the appropriated biocatalyst to be employed. Microbial
present as minor components in the final product, however, they may cells are more advantageous than enzymes since they have higher sta-
represents up to 50% of the total costs (Fanaro et al., 2016). In 2016, bility and there is no need of co-factors addition for the reaction to
the global sales of aromas and fragrances was approximately US$24.5 occur. Microorganisms also show high grow rates, it is easy to manip-
billion, being the companies Givaudan, Firmenich and IFF responsible ulate fermentation parameters and perform genetic engineering in
for 18.7%, 13.5% and 12.3% of the market share, respectively order to increase strain performance and can be applied in large-scale
(Leffingwell and Associates, 2017). It is expected an annual grow rate of processes (Rodrígues-Bustamante and Sánches, 2007).
5–6% for this market (UBIC Consulting, 2014). Generally, in screening assays, the first experiments aim to identify
Nowadays, the major production of aroma compounds is from the most adequate and robust strain for each process, since both sub-
chemical methods, since direct extraction from nature can result in strate and product may cause inhibitory effects on cell growth.
complex mixtures, with low concentrations and are very restrict to Therefore, all strains to be selected are incubated with the substrate to
geographical availability and seasonality (Longo and Sanromán, 2006; be used in order to find which strains can use the substrate as sole
Berger, 2015). The growing awareness on health and nutrition along carbon source, indicating the existence of a metabolic pathway for
with the concern of having synthetic chemicals added to food have substrate degradation and possible accumulation of interesting sec-
encourage efforts in the development of biotechnological processes for ondary metabolites (Bicas and Pastore, 2007). It is interesting to notice
obtaining flavor compounds that can be labeled as “natural” (Bicas that in this first step, the incubation conditions are usually similar in
et al., 2009; Maróstica Jr. and Pastore, 2007). various screening works. This fact may be due to the necessity to
Industrially, some companies are developing methods for bio- maintain the optimum temperature, pH and agitation for strain growth
technological aroma production. The Amyris Company produces β- in order to have a better comparison of the results. The resistance to
farnesene from sugarcane, using a genetically modified strain of process conditions and ability of substrate degradation are important
Saccharomyces cerevisiae. This sesquiterpene can also be employed in biocatalysts characteristics; however, it does not guarantee high con-
aroma industry as precursor for other aroma compounds production centration or specificity of products. Thus, further investigation is ne-
(Felipe et al., 2017; Schempp et al., 2017). The same company, in a cessary to evaluate the compounds formed after incubation. Usually,
partnership with Firmenich, is the responsible for the production of a gas chromatography is used to identify (gas chromatography coupled
patchouli oil substitute, named Clearwood, with better sensorial qua- with mass spectrometry) and quantify the volatile compounds obtained
lities than the natural oil (Leffingwell and Leffingwell, 2015). The (Bier et al., 2011).
biotechnological production of valencene has been reported by Evolva In this context, many researches have been focusing in identify new
and Isobionics companies. This compound can also be used as precursor microorganisms and enzymes that can be used in biotechnological
for oxidation to nootkatone, which is also market by these companies to process, through isolation, screening and selection of the best strains
be applied as flavor as well as potential natural insect repellent capable to produce aroma compounds from different substrates. This is
(Schempp et al., 2017). In their processes, Isobionics uses a strain of a promising methodology since is a natural way to obtain new products
Rhodobacter sphaeroides which contains genes from the terpene bio- at low costs (Rottava et al., 2010). Table 1 summarizes the screening
synthesis pathway (Huembelin et al., 2014). studies for aroma compounds production.
Microorganisms and enzymes can be employed as catalysts in two Rottava et al. (2010) tested 400 microorganisms (fungal, bacteria
different processes: de novo synthesis or biotransformation of natural and yeasts) for biotransformation of R-(+)-limonene and (−)-β-pinene
precursors. For the first case, only simple nutrients (sugars, alcohols) in a three-step process. Firstly, all strains were incubated in minimal
are present in fermentation media and the microorganisms utilize a medium containing only terpene as carbon source to verify which
broad array of enzymes and metabolic pathways to obtain a mixture of strains were able to use these substrates to grow. In the second step, the
various compounds as final product (Molina et al., 2012). With this 190 strains that could grow in the presence of R-(+)-limonene or
methodology, it was already reported the production of ethyl acetate (−)-β-pinene were screened for their ability to produce α-terpineol,
and ethyl hexanoate using Kluyveromyces marxianus and Neurospora sp., resulting in 4 strains selected for biotransformation of β-pinene. The
321
Table 1
Studies of screening/isolation of microorganism for aroma compounds production.
Substrate Microorganisms source Microorganisms isolated Optimized process conditions Product Reference
M.G. Pessôa et al.
a a
(-) β-pinene 400 commercial and isolated strains Aspergillus niger ATCC 16404, 35 °C, without vitamin solution addition α-terpineol (2856.54 ± 50.23 mg/L) and Rottava et al. (2010)
b
A. niger ATCC 9642, fenchol (traces),
c b
A. niger ATCC 1004, α-terpineol (688.13 ± 41.27 mg/L),
d c
Penicillium camembertii ATCC α-terpineol (172.07 ± 32.94 mg/L),
d
4845 α-terpineol (24.38 ± 2.78 mg/L) andtrans-
pinocarveol (traces)
R-(+)-limonene Citric fruit orchards and industries 05.01.35 1.75% of substrate, 2 g inoculum and 1:1 α-terpineol (1700 mg/L) Rottava et al. (2011)
ethanol: substrate ratio
R-(+)-limonene and (-) β-pinene Laboratory collection or isolated from 5, 16, 3a and Y4b Not optimized Sabinene, carveol, dihydrocarveol, carvone, Bier et al. (2011)
reforestation area perillyl aldehyde, perillyl alcohol, guaiol and
others
a a
R-(+)-limonene Endophytic from Pinus taeda Phomopsis sp Mineral media with terpene carvone (536 mg/L), limonene-1,2-diol Bier et al. (2016)
b
Orange residue extract (2.08 g/L) and α-terpineol (23.64 mg/L)
b
carvone (12.56 mg/L), limonene-1,2-diol
(2.10 g/L) and α-terpineol (34.22 mg/L)
R-(+)-limonene Citrus processing plant and orange, lime and 238 strains isolated (not YM media, 0.1% of substrate, 30 °C and 70 strains able to grow using limonene as single Bicas and Pastore
mint identified) 150 rpm carbon source (2007)
Products were not identified in this study
a a
α-pinene Samples collected in the Arctic tundra Chrysosporium pannorum A-1 1.5% of α-pinene, 10 °C, basal medium verbenol (722 mg/L) Trytek et al. (2015)
b b
1% of α-pinene, 20 °C, basal medium verbenone (176 mg/L)
(R)-(+)-citronellol 60 fungal strains Aspergillus sp. Not optimized, SPME method used in the cis- rose oxide (54%) Demyttenaere et al.
Penicillium sp screening trans- rose oxide (21%) (2004)
nerol oxide (12%)
a a
(R)-(+)-citronellol Mango (Mangifera indica) LB-2025 (Penicillium sp.) Induction of microorganisms prior rose oxide (4.5 mg/L) Molina et al. (2013)
322
b b
LB-2036 (Penicillium sp.) biotransformation process rose oxide (6.5 mg/L)
a a
R-(+)-limonene Mango and acerola (Malpighia emarginata) LB-2025 (Penicillium sp.) Induction of microorganisms prior α- terpineol (10 mg/L)
b b
LB-2038 (Aspergillus sp.) biotransformation process Carvone (47 mg/L)
Geraniol Sapoti (Manilkara zapota) LB-2041 Not optimized 4-methyl-3-penten-1-ol (97% of similarity) and
6-methyl-5-hepten-2-ona (93% of similarity)
a a
(S)-(+)-Linalool Graviola (Annona muricata) LB-2077 Not optimized Geraniol (7 mg/L)
b b
LB- 2010 α-terpineol (150 mg/L)
c c
LB-2050 α-terpineol (130 mg/L)
a a
(+)- valencene Culture collection Botryodiplodia theobromae Biphasic system, not optimized 231.7 ± 2.1, Palmerín-Carreño et al.
b
1368, 216.9 ± 5.8 (2015)
b c
Yarrowia lipolytica 2.2ab 100.8 ± 2.6 mg/L
c
Phanerochaete chrysosporium
Molasses and L-phenylalanine Culture collection Kluyveromyces marxianus CBS Addition of oleyl alcohol as second phase 2-phenylethanol (3 g/L) Etschmann et al. (2003)
600 and CBS 397 and 35 °C
Isoeugenol Soil samples Bacillus fusiformis CGMCC1347 Addition of a resin HD-8 to remove vanillin Vanillin (8.1 g/L) Zhao et al. (2006)
and avoid product inhibition
Isoeugenol Soil samples from greenhouses Bacillus pumilus S-1 Addition of isoeugenol in the stationary Vanillin (3.74 g/L) Hua et al. (2007)
phase to avoid inhibition and lower its
toxicity
Isoeugenol Soil samples from greenhouses of basil Bacillus subtilis B2 Not optimized Vanillin (0.9 g/L) Shimoni et al. (2000)
(Ocimum basilicum) and Tarragon (Artemisia
drancunculus)
Isoeugenol Soil samples Pseudomonas putida I58 Basal media with 5 g/L of isoeugenol Vanillic acid (98% conversion) Furukawa et al. (2003)
Ferulic acid Orchards soil samples Streptomyces sp. V-1 Fed-batch mode, with addition of 5 g/L of Vanillin Xu et al. (2009)
ferulic acid, up to 45 g/L
Molasses Sea sediment samples Bacillus subtillis DL01 Not optimized Acetoin (76 g/L) Dai et al. (2015)
The superscript letters are used to correlate the experiments performed or the microorganisms used with the results obtained. The results are referent to the microorganism or experiments with the same letter.
third step comprised the evaluation of some process variables effects on value flavor compounds. The authors screened 60 fungal strains using
α-terpineol production. It was show that the highest concentration solid-phase microextraction (SPME) to monitor product formation. It
(15,494.34 mg/L ± 193.87) was reached by Aspergillus niger was found that Aspergillus sp. and Penicillium sp. were able to produce
ATCC16404 when incubated at 35 °C, without addition of vitamins. cis- and trans-rose oxides and nerol oxide (up to 54, 21 and 12%, re-
Later, the same group isolated several strains from citric fruit spectively). It is interesting to highlight the SPME employed in this
orchards and industries and performed and optimization of α-terpineol work is a fast and solvent-free alternative to perform screening of
production. Through a central composite rotatable design (CCRD) with strains even from solid cultures in a non-destructive way.
tree independent variables, it was possible to obtain a maximum α- Some Brazilian fruits are rich in monoterpenes, therefore, Molina
terpineol concentration of 1700 mg/L using the strain 05.01.35. The et al. (2013) selected fungal strains capable of growing in fruits, such as
fermentation conditions were: R-(+)-limonene concentration of 1.75%, guava, acerola, mango, passion fruit, tamarind, star fruit, papaya,
mass of inoculum of 2 g and substrate to ethanol volume ratio of 1:1. umbu, sapoti, graviola and seriguela. A total of 36 fungal strains were
When the same conditions were applied in (−)-β-pinene bio- tested in biotransformation of the monoterpenes citronellol, limonene,
transformation, about 770 mg/L of α-terpineol was reached (Rottava linalool and geraniol (Table 1). The isolated strains with the highest
et al., 2011). α-Terpineol is an interesting product due its characteristic product concentration were LB-2025 and LB-2036, with the production
lilac odor, with a sweet taste and can be used in formulation of bev- of rose oxide from citronellol and LB-2025 and LB-2038 for production
erages, baked goods and candies (Burdock, 2010). of α-terpineol and carvone respectively, from limonene. The authors
Bier et al. (2011) also selected microorganisms for biotransforma- also performed a study of cell induction in an attempt to increase the
tion of terpenic substrates. In this study, from 41 strains tested, 21 were final product concentration for the selected strains. It was shown a 5-
isolated from reforestation area (Paraná, Brazil) and 20 obtained from and 8-fold increase in rose oxide concentration for strains LB-2025 and
Culture Collection of the Biotechnological Process Laboratory (Federal LB-2036, respectively. In contrast, the induction did not have effects in
University of Paraná). All strains were tested for terpene resistance by the biotransformation of limonene. When geraniol was used as sub-
growing in PDA medium containing 1% of limonene or 1% of α- and β- strate, a strain identified as LB-2041 isolated from Sapoti was able to
pinene (1:1), and for product formation after incubation (30 °C, produce 4-methyl-3-penten-1-ol and 6-methyl-5-hepten-2-ona (not
120 rpm) in mineral medium with terpenes limonene, α-pinene or β- quantified). The strains LB-2077, LB-2010 and LB2050 were able to
pinene as sole carbon source, for 7 days. The results show that after this convert (S)-(+)-linalool into geraniol (7 mg/L), α-terpineol (150 mg/L)
period, four strains (5, 16, 3a and Y4b) were able to convert limonene and α-terpineol (130 mg/L).
into interesting products, such as carveol, carvone, perillyl alcohol and Bicas and Pastore (2007) performed an isolation of microorganisms
others. For pinenes biotransformation, only the strain Y4b was able to resistant to limonene containing environments. This is a good indica-
produce α-fenchene and sabinene using β-pinene as substrate, but no tion that these microorganisms may be able to metabolize limonene and
product was observed from α-pinene biotransformation. use it as precursor in the production of several compounds. In this
The ability of an endophytic fungus in the biotransformation of process, samples collected in a citrus processing plant (Araras, SP –
terpenes has been reported. A strain identified as Phomopsis sp. was Brazil) or vegetables (orange, lime and mint) obtained in the local
isolated from Pinus taeda and proved to be resistant to limonene. It was market were incubated at 30 °C and 150 rpm, in YM media (1% glucose,
able to produce carvone, carveol, α-terpineol, terpinen-4-ol and limo- 0.5% peptone, 0.3% malt extract, 0.3% yeast extract) with 0.1% (v/v)
nene-1,2-diol when cultivated for 10 days at 30 °C and 120 rpm, being of limonene for 48 h or 7 days. A total of 238 strains were isolated, but
that the product concentration was variable according to the carbon only 70 were able to grow in mineral media containing only limonene
source used. When cultivated in mineral medium with terpenic sub- as carbon source. This isolation process is the first step for bioflavors
strate addition in a single fed-batch mode, 536 mg/L of carvone, 2.08 g/ production using terpenic substrates and the biotransformation process
L of limonene-1,2-diol, 89.74 mg/L of terpinen-4-ol and 23.64 mg/L of should also be evaluated in order to identify the compound that can be
α-terpineol were obtained, whereas when cultivated in orange residue obtained.
extract, the concentration of carvone and terpinen-4-ol decreased to In addition to monoterpenes, sesquiterpenes can also be used as
12.56 mg/L and 17.01 mg/L respectively and α-terpineol increased to substrate in biotransformation assays. (+)-Valencene is the main by-
34.22 mg/L (Bier et al., 2016). The use of endophytic microorganism ciclic sesquiterpene found in orange peel oil and can be employed as
may be advantageous due its ability to produce a great variety of me- precursor in (+)-nootkatone production, therefore, Palmerín-Carreño
tabolites, which can be isolated and characterized as bioactive com- et al. (2015) performed a screening with six strains from culture col-
pounds with potential for use in industry, agriculture and medicine lections to produce (+)-nootkatone. It was also tested the influence of
(Pimentel et al., 2011; Strobel and Daisy, 2003). different bioconversion systems: aqueous, organic and biphasic, using
The psychotrophic fungus Chrysosporium pannorum A-1, isolated orange essential oil as organic phase. The results show that nootkatone
from soils samples collected in the Arctic tundra, proved to be able to concentrations of 231.7 ± 2.1, 216.9 ± 5.8 and 100.8 ± 2.6 mg/L
produce verbenol and verbenone through α-pinene biotransformation. were reached using Botryodiplodia theobromae 1368, Yarrowia lipolytica
The maximum growth of this strains was observed when the fungus was 2.2ab and Phanerochaete chrysosporium respectively, when cultivated at
incubated at 20 °C, 150 rpm in a basal medium (malt extract 1%, pep- 30 °C, 200 rpm for 4 days. There were obtained similar results in single
tone 0.5%, glucose 1%, and yeast extract 0.5%), although the tem- aqueous phase, single organic phase and biphasic experiments, showing
perature of incubation could vary from 5 to 20 °C. The highest verbenol that the method for strain cultivation did not affect valencene bio-
production (722 mg/L) was obtained using a 72 h-old mycelium in a transformation, although both substrate and product can cause growth
medium containing 1.5%(v/v) of α-pinene, incubated at 10 °C, whereas inhibition in high concentrations.
verbenone production was optimized with a 48 h-old inoculum, 1% of In a different approach, a molasses-based media with addition of L-
substrate and incubated at 20 °C, reaching 176 mg/L. The authors first phenylalanine was employed in production of 2-phenylethanol. This
reported the use of a psychotrophic fungus in biotransformation process compound has a rose-like odor, with an initially bitter taste and re-
as well as the conduction of the experiments in temperatures below miniscent of sweet and peach and is an important raw material in
25 °C. These conditions are suitable for aroma production since the fragrance industry (Burdock, 2010; Etschmann et al., 2003). From 14
volatile compounds can be lost due to evaporation at high tempera- strains, Kluyveromyces marxianus CBS 600 and CBS 397 showed the
tures, therefore, a high amount of products can be recovered (Trytek highest production of 2-phenylethanol and proved to be the most
et al., 2015). thermo tolerant strains, reaching 1 g/L of product. Although promising,
Demyttenaere et al. (2004) described for the first time the bio- it is necessary to overcome product inhibition during fermentation
transformation of the acyclic monoterpene (R)-(+)-citronellol into high process. Thus, an organic phase constituted by oleyl alcohol was
323
employed for in situ product removal, leading to a final 2-phenylethanol of production volatile sulfur compounds (VSC). These compounds are
concentration of 3 g/L. present in ripened cheeses, providing its characteristics flavors, and are
Vanillin is one of the main aroma compounds, with a total market produced from methanethiol (MTL), a degradation product of L-me-
value of US$ 600 million and total volume of approximately 18 metric thionine. The method is based in the use of a double-layer petri dish,
tons, being that only a small amount of this total corresponds to natural being the upper layer constituted of a chemical for detection of free
vanillin (Leffingwell and Leffingwell, 2015). It is estimated that ap- thiols, therefore, MTL production can be quantitated by measuring the
proximately 500 kg of Vanilla planifolia pods are necessary to obtain yellow-orange color intensity. Through this methodology, 18 strains of
1 kg of natural vanillin, therefore, its biotechnological production is Geotrichum candidum strains isolated from cheese could be selected for
advantageous (Felipe et al., 2017). Several microorganisms isolated thiol production, and the compounds obtained were further correlated
from soil samples are reported as able to convert isoeugenol to vanillin, with VSC production using gas chromatographic analysis (Guichard and
such as Bacillus fusiformis (final concentration of 8 g/L of vanillin), Bonnarme, 2005).
Bacillus pumilus (3.75 g/L), Bacillus subtilis (0.9 g/L) and Pseudomonas Another technique to select cheese-related bacteria and the aroma
putida, which converts isoeugenol to vanillic acid (98% conversion rate) produced was developed recently. It was used a curd-based medium for
(Zhao et al., 2006; Hua et al., 2007; Shimoni et al., 2000; Furukawa strain growth and aroma production in order to mimic cheese manu-
et al., 2003). facture, a headspace trap method coupled to gas chromatography-mass
Nowadays, biotechnological vanillin is produced industrially by spectrometry for samples analysis and a metabolomics-based method of
several companies, such as Evolva-IFF, Mane, Solvay and BASF, most of data processing. This methodology was used for screen aroma pro-
them employing genetic modified organisms (Felipe et al., 2017). The duction of 11 different species and, in total, 52 volatile compounds
Chinese company Shanghai Apple Flavor & Fragrance Group Co. Ltd. were identified, being ethyl esters, sulfur compounds, branched-chain
uses a wild-type strain isolated from orchards soil samples to convert alcohols, branched chain acids and others, the main compounds
ferulic acid into vanillin. In this process, a strain named as Streptomyces (Pogacic et al., 2015). The same approach was used to evaluate inter-
sp. V-1 is cultivated in GY medium (glucose 15 g/L, yeast powder 8 g/L, species and intraspecies diversity in production of cheese aroma com-
magnesium sulfate 0.8 g/L, sodium chloride 2 g/L (pH 7.2)) for biomass pounds by Propionibacterium sp. (Yee et al., 2014).
growth and then 5 g/L of ferulic acid is added for biotransformation Due the great importance of aroma compounds in product for-
until substrate concentration reaches 45 g/L. The culture is incubated at mulation and its impact on consumers’ preference, it is extremely ne-
30 °C, 120 rpm for 24 h (Xu et al., 2009). This example shows the reality cessary to develop and find new volatile compounds to be added into
and feasibility of employing isolated strains in industrial biotechnolo- food, cosmetics and pharmacy. To select new isolated microorganisms
gical processes for value-added compounds. is an interesting approach since is a low cost technique and a new range
Dai et al. (2015) isolated a strain named Bacillus subtillis DL01 from of metabolites can be produced and employed as flavoring agents.
sea sediment samples which as able to produce acetoin in a simple These screening steps are, in general, very laborious since a large
fermentation media: molasses, (NH4)2SO4 and MnSO4. The best results number of biocatalysts can be obtained after isolation procedure and,
of 76 g/L acetoin, productivity of 1 g/L.h and yield of 0.421 g/g could up to this moment, there are few methodologies for fast identification
be obtained when initial sugar concentration in molasses was 210 g/L, and quantification of volatile compounds, such as the colorimetric assay
showing that the strain has high sugar tolerance and low oxygen re- developed by Guichard and Bonnarme (2005). Therefore, many efforts
quirement. Acetoin is mainly used as food additive with application in must be made is this area, aiming to facilitate strain isolation using high
butter, milk, yogurt or strawberry flavors (Burdock, 2010). throughput systems and to develop and improve new feasible meth-
Aroma compounds can also be derivatives from carotenoids. The odologies for aromas production, identification and analysis.
cleavage of these compounds may lead to the formation of norterpe-
noids/norisoprenoids such as ionones and damascones, that contribute 2.2. Biosurfactants
as aroma in tea, grapes, roses and wine. In this context, it was per-
formed a screening of microorganisms with ability to use lutein as Biosurfactants are natural compounds with surface and emulsifying
substrate for tobacco aroma production and to degrade β-carotene into activities and can be produced by several microorganisms including
β-ionone, β-cyclocitral, dihydroactinidiolide, and 2-hydroxy-2,6,6-tri- bacteria, filamentous fungi and yeasts. They are considered an eco-
methylcyclohexanone. For the initial phase of the screening assays, an friendly alternative to the use of synthetic surfactants due to its suitable
extract composed of 89% of lutein was added in a defined solid characteristics of biodegradability, low toxicity and stability (Banat
medium, were the microorganisms were inoculated. It was observed the et al., 2014a; Cameotra and Makkar, 2010; Soberón-Chávez and Maier,
ability of 19 colonies to grow using lutein as carbon source as well as 2011; Mnif and Ghribi, 2015). Structurally, these microbial metabolites
the pigment degradation by the formation of a colorless halo around the consist in amphiphilic compounds formed by hydrophilic and hydro-
colony. Further experiments show that only 2 colonies were able to phobic moieties and are classified in two major groups according of
produce volatile compounds being the best results obtained in a com- molecular weight: low or high molecular weight biosurfactants; sub-
bination of a bacteria (Paenibacillus amylolyticus) and a yeast divided in different subgroups and families depending of their struc-
(Trichosporon asahii) (Rodrígues-Bustamante and Sánches, 2007). tural characteristics (Gudiña et al., 2013; Mnif and Ghribi, 2015;
The genetic diversity among the microorganisms, even from the Paulino et al., 2016).
same genera, can lead to the formation of broad range of aroma com- The most studied biosurfactants are those belonging to the low
pounds. The exploration and understanding of this diversity provide molecular weight (LMW) group, represented by glycolipids and lipo-
tools for improvement and development of strains with favorable peptides, that exhibit good stability in several pH, salinity and tem-
phenotypes for aroma production. After a complete study on 301 perature conditions and have been described for important potential
Saccharomyces sp. strains and their potential for aroma compounds applications in food, medical, cosmetic, agricultural and environmental
production, it was possible to select parental strains and obtain superior fields (Fig. 3) (Mnif and Ghribi, 2015; Paulino et al., 2016; Banat et al.,
hybrids with great potential for production of mainly isoamyl acetate 2014b; Joy et al., 2017). Moreover, among the glycolipid biosurfac-
and ethyl acetate from glucose when incubated at 30 °C for 7 days. tants, rhamnolipids, sophorolipids, trehalolipids and mannosylery-
These new strains are considered non-GMO and, according with the thritol lipids are the most important groups, while surfactins, fengycins,
authors, can be readily applied in industry (Steensels et al., 2014). and iturins are the best know lipopeptides.
The development of a feasible methodology for large-scale The high molecular weight (HMW) biosurfactants are considered
screening of microorganism is another challenge to be overcome. A efficient emulsifying agents and its application in bioremediation as
plate technique was developed in order to easily identify strains capable well as to remove hydrophobic compounds from different environments
324
Fig. 3. Low molecular weight biosurfactants with potential applications in food, medical, cosmetic, agricultural and environmental fields.
have been reported (Pacwa-Płociniczak et al., 2011). However, some et al., 2013). In order to overcome these problems, different approaches
authors have used the term bioemulsifier to classify these amphiphilic have been suggested such as the application of statistical methods to
compounds, emphasizing their different physico-chemical properties optimize the process conditions, use of alternative substrates, new
and physiological roles, requiring different screening methods purification strategies using sustainable solvents, genetic engineering
(Uzoigwe et al., 2015). Considering the fact that some compounds have tools, and screening studies for prospection of novel microorganisms
surface activity and emulsifying properties, the classification of bio- from different environments able to produce of high yields of bio-
surfactants based on molecular weight has still been used in the lit- surfactant (Zhao et al., 2016; Ebadipour et al., 2016; Bhangale et al.,
erature. 2014; Tavares et al., 2013; Mukherjee et al., 2006; Cai et al., 2014;
In this context, the group of HMW biosurfactants comprises dif- Kennedy et al., 2011).
ferent compounds including amphipathic polysaccharides, lipopoly- Many studies have been performed to isolate novel microorganisms
saccharides, proteins, lipoproteins or mixtures of these biopolymers, for biosurfactant production and different screening assays for identi-
named by some studies as polymeric biosurfactants (Gudiña et al., fication of promising strains can be used for this purpose. Among the
2013; Ron and Rosenberg, 2001; Pacwa-Płociniczak et al., 2011). The main qualitative methods used in screening of microbial biosurfactant
most studied HMW biosurfactants are Emulsan and Biodispersan pro- producers, surface and interfacial tension measurement, oil displace-
duced by Acinetobacter calcoaceticus strains, Alasan produced by ment test, drop collapse method, emulsification index, CTAB (N-Cetyl-
A. radioresistens, Liposan produced by Candida lipolytica, and others N,N,N-trimethylammoniumn bromide) agar plate method, hemolytic
(Zosim et al., 1982; Rosenberg et al., 2001; Toren et al., 2001; activity, and BATH (bacterial adherence to hydrocarbons) assay are the
Cirigliano and Carman, 1984). most used (Pizon and Ju, 2009; Desai and Banat, 1997; Walter et al.,
From industrial point view, many patents were filled regarding the 2010; Cai et al., 2014). Each test is based into evaluating the surface
application of biosurfactants for different purposes, including its addi- activity or emulsifying properties of the microbial metabolites pro-
tion in detergent and cosmetic formulations, enhance fluid recovery, duced after or during cultivation by wild microorganism isolated.
protection against corrosion, biopesticides, biodispersants, and others Moreover, the use of natural samples as an isolation source is promising
(Randhawa and Rahman, 2014; Ferreira et al., 2017; Paulino et al., and it has been reported the use of contaminated or soil from extreme
2016; Almeida et al., 2017). In addition, the expected biosurfactant environments, leaves, fruits, honey, sugarcane, wastes and even insect
market in 2020 is approximately 2.3 billion dollars and a production of gut (Bodour et al., 2003; Khopade et al., 2012; Gesheva et al., 2010;
approximately 462 thousand tons, showing the growing commercial Elazzazy et al., 2015; Ohmart et al., 1988; Sabaté et al., 2009).
interest in these biotechnological products (Focus on Surfactants, 2014; Table 2 comprises recent studies focused in isolation and screening
Grand View Research Inc., 2014). The main companies focusing in of novel biosurfactant-producers and the natural sources used as a
biosurfactant production or its application are Ecover, Henkel, Kaneka sample for isolation. However, screening studies often report the bio-
Co., AGAE technologies LCC, Jeneil Biosurfctant Co. LLC, Synthezyme diversity of microorganisms found and few data about yield and con-
LLC, Soliance, MG Intobio Co. Ltd and Saraya Co. Ltd (Randhawa and centration of the biosurfactant produced are provided. This fact is jus-
Rahman, 2014). However, the industrial production and insertion of tified because generally the most promising producers will be later
biosurfactants in market is limited by factors as high costs of the culture studied as the best conditions for conducting the biotechnological
media used in biotechnological process, complexity in purification steps process to biosurfactant production.
that many times employed diverse organic solvents and acids, and the The differential metabolic and physiological capabilities of marine
low yields achieved in some processes (Franzetti et al., 2010; Mnif microorganisms have been described as a determinant factor for their
325
M.G. Pessôa et al.
Table 2
Novel biosurfactant-producers isolated from natural sources and their potential applications.
Biosurfactant type Biosurfactant family Microorganism Isolation source Potential application Reference
Glycolipid Rhamnolipid Rhodococcus fascians Antarctic soil Antibacterial agent Gesheva et al. (2010)
Acinetobacter junii Oil–water mixture samples from Enhance oil recovery Dong et al. (2016)
petroleum reservoir
Pseudomonas aeruginosa M408 Petroleum-contaminated soil Emulsifying agent Ji et al. (2016)
Pseudomonas aeruginosa strains Hydrocarbon-contaminated soil n.d Rikalovic et al. (2013)
Serratia rubidaea SNAU02 Hydrocarbon-contaminated soil Emulsifying agent Nalini and Parthasarathi (2014)
Pseudomonas nitroreducens Petroleum-contaminated soil Enhance oil recovery Onwosi and Odibo (2012)
Achromobacter sp. (PS1) Oil spilled refinery soil Bioremediation processes Joy et al. (2017)
Bacillus sp. (SLDB1) Tank settled refinery sludge
Sophorolipids Wickerhamiella domercqiae CGMCC Oil-containing waste water sample Pharmaceutical and petroleum indutries Ma et al. (2012)
1576
Pseudozyma churashimaensis leaves of Saccharum officinarum n.d Morita et al. (2011a)
Mannosylerythritol Lipid Ustilago scitaminea NBRC 32730 Sugarcane juice Ceramide-like skin-care property Morita et al. (2011b)
Pseudozyma tsukubaensis leaves of Perilla frutescens Efficient production of the diastereomer MEL-B Morita et al. (2010)
from vegetable oils
Lipopeptide Surfactin Bacillus subtilis LSFM-05 Hydrocarbon contaminated soil Emulsifying agent Faria et al. (2011)
Bacillus subtilis C4, M1, and G2III honeybee gut and honey samples Antimicrobial agent against honeybee pathogens Sabaté et al. (2009)
326
strains
Aneurinifactin Aneurinibacillus aneurinilyticus SBP- Sediment samples from coastal sites Antimicrobial agent Balan et al. (2017)
11
Pontifactin Pontibacter korlensis SBK-47 Petroleum contaminated sea water Antimicrobial and anti-adhesive agent Balan et al. (2016)
Mixture of surfactin, fengycin, and Pseudomonas sp. WJ6 Heavy oil-contaminated soil Heavy oil sand washing Xia et al. (2014)
lichenysin
n.d Nesterenkonia sp. MSA31 Marine sponge Fasciospogia cavernosa Antioxidant and antimicrobial agent Kiran et al. (2017)
tissue
a
n.d Achromobacter sp. HZ01 Crude-oil contaminated seawater Emulsifying agent Deng et al. (2016)
n.da Bacillus pseudomycoides BS6 Long-term contaminated soil with edible Emulsifying agent Li et al. (2016)
oil
n.d Bacillus brevis sediment of mangrove trees Emulsifying agent Mouafi et al. (2016)
Different isoforms of surfactin, iturin and Bacillus methylotrophicus DCS1 Hydrocarbon contaminated soil samples Potential for enhancing solubility of hydrocarbon Jemil et al. (2016) and Jemil et al.
fengycin (2017)
n.d Staphylococcus xylosus STF1 Petroleum contaminated soil samples Antimicrobial agent Keskin et al. (2015)
n.d Ochrobactrum sp. (GREW1) Refinery raw oil effluent Bioremediation processes Joy et al. (2017)
n.d Bacillus sp. (SB2) Desert soil
n.d: not described.

a
A novel cyclic lipopeptide.
potential in secondary metabolites of industrial interest, including after 144h the cell free supernatants obtained by centrifugation were
biosurfactants, and in the last years many studies were conducted for tested using drop collapsing, oil displacement, lipase, haemolytic ac-
prospection of biosurfactant producers using samples from marine en- tivity, surface tension measurement and emulsification index. The
vironments (Kennedy et al., 2011; Satpute et al., 2010; Das et al., 2010; biosurfactant produced by Nesterenkonia sp. MSA31 strain showed
Deepika et al., 2016). promising characteristics such as a low surface tension value (34 mN/
Cai et al. (2014) performed a screening study with marine micro- m), high emulsification activity (75%), 8 mm of clear diameter in oil
organisms isolated from contaminated sea water and sediment samples displacement method, a flat drop in drop collapsing test and a CMC
from North Atlantic Canada coastal line. In this work, 55 strains be- value of 18.6 μg/mL. Furthermore, the lipopeptide produced in this
longing to Bacillus sp., Rhodococcus sp., Acinetobacter sp., Halomonas sp., study exhibited non-toxic characteristic in the brine shrimp cytotoxicity
Alcanivorax sp., Pseudomonas sp., Streptomyces sp. and Exiguobacterium assay, antioxidant and antimicrobial potential, and its incorporation in
sp. genera were isolated and tested for surface and emulsifying activ- food products (muffin) improved organoleptic qualities as texture and
ities. The best results of surface tension reduction were observed to color.
Bacillus sp. and Rhodococcus sp., that exhibited values around Considering the importance of low molecular weight biosurfactants
36–31.3 mN/m. In addition, high emulsification index (E24 > 50%) due to their attractive surface and interfacial properties, novel lipo-
were obtained to 3 Acinotobacter sp. and two Rhodococcus sp. strains. peptides biosurfactants could be produced by microorganisms isolated
Similarly, Hassanshahian (2014) used sediments and seawater samples from marine environments, for example Pontibacter korlensis SBK-47,
collected from the coastal line of Bushehr provenance (Persian Gulf) to Bacillus circulans, Bacillus licheniformis, Brevibacterium aureum MSA13,
isolate 14 bacteria strains and screened for biosurfactant production Nocardiopsis dassonvillei MAD08, and others (Balan et al., 2016; Balan
capacity by different screening tests, including hemolysis in blood agar, et al., 2017; Das et al., 2008; Lawrance et al., 2014; Kiran et al., 2010;
oil spreading drop collapse, BATH and emulsification activity. The Selvin et al., 2009; Gandhimathi et al., 2009). However, the production
emulsification activity and BATH observed to microorganisms in this of novel lipopeptides also have been reported for microorganisms iso-
study ranged from 10% to 65% and 1.7% to 59%, respectively, while lated from other environments, including oil contaminated soil samples
the surface tension values measured were of 50.2–34.5 mN/m. (Faria et al., 2011; Li et al., 2016).
The amount of potential biosurfactant producers identified in The production of surfactin by Bacillus sp. strains can be considered
screening studies can be variable according to type and number of as- the most studied approach found in literature about the lipopeptide
says applied for this purpose. Using the same approach of the studies biosurfactant production. In this context, the production of 1.36 g/L of
mentioned above, Mohanram et al. (2016) used nine different methods surfactin by B. subtilis LSFM-05 isolated from oil contaminated soil
of screening to isolate 19 bacterial strains from oil-polluted seawater sample was reported (Faria et al., 2011). The biosurfactant was purified
samples collected of three Mumbai Harbor stations. In this case, after from 0.5 L of foam collected during 72 h of process and exhibited sur-
isolation step, each bacterial strain was grown in flasks contained face tension activity varying of 29.5–32.1 mN/m, while the emulsifi-
100 mL of modified Bushnell–Haas broth (MBHB) mineral media with cation index after 24 h (EI 24) ranged of 49.6–67%.
1% of high-speed diesel as a carbon source, incubated at 30 °C, 180 rpm The production of pontifactin by a new marine bacterium identified
for 3 days. Then, the culture broth was centrifuged and the supernatant as Pontibacter korlensis isolated from the coastal waters was described
that was used to perform the drop-collapse assay, oil spreading assay, by Balan et al. (2016). The biotechnological process was carried using
microplate assay, penetration assay, stable emulsification assay, and glucose mineral salts medium prepared in sea water, and the isolated
surface tension measurement. Also, the bacterial cells were used in the strain was incubated at 37 °C, 150 rpm for 90 h reaching a maximum
blue agar method, hemolytic and BATH assays. The promising bio- yield of 2.34 g/L. The lipopeptide produced exhibited good surface and
surfactant producers (19 bacterial strains) were those that showed po- emulsifying activities, with surface tension reduction and CMC value of
sitive response to more than four screening assays. Moreover, surface 25 mN/m and 25 mg/L, respectively. These authors also proved the
tension values obtained ranged from 52.7 mN/m to 27.6 mN/m, with stability of this biosurfactant to wide range of pH and temperature, and
E24 index greater than 70% up to 20%. The phylogenetic character- its promising antimicrobial and anti-biofilm activities against several
ization of bacterial isolates revealed ten different genera, among them, pathogens such as Streptococcus mutans, Micrococcus luteus, Salmonella
Alcanivorax sp., Acinetobacter sp., Bacillus sp., Comamonas sp., Halo- typhi, Klebsiella oxytoca, Bacillus subtilis, Staphylococcus aureus, and Vi-
monas sp., Marinobacter sp., Pseudomonas sp, and Serratia sp., being that brio cholerae.
for first time it was described strains belonging Nesterenkonia sp. and Other recent study reported the production of a novel lipopeptide
Chryseomicrobium sp. genera as marine biosurfactant producers. biosurfactant by a marine bacterium Achromobacter sp. HZ01 isolated
Different culture media are used in the screening of biosurfactants from crude-oil contaminated seawater (Deng et al., 2016). When cul-
producers and the composition of these media varies greatly depending tivated in fermentation medium containing glycerine (40 g/L) as a
on the type of microorganism and biosurfactant produced. The pro- carbon source at 28 °C, 150 rpm for 144 h, it was possible to obtain a
duction of sophorolipids was described using a Wickerhamiella do- maximum surfactant concentration of 6.84 g/L, presenting a surface
mercqiae strain, isolated from an oil-containing waste water sample, tension reduction of 24.2 mN/m. Furthermore, this biosurfactant ex-
when cultivated in fermentation medium composed by glucose (80 g/ hibited high emulsifying activity against different mineral and vege-
L), yeast extract (3 g/L), KH2PO4 (1 g/L), Na2HPO4·12H2O (1 g/L), table oils as a soybean oil, sunflower oil, olive oil, kerosene and diesel
MgSO4·7H2O (0.5 g/L) and oleic acid (60, v/v) at 30 °C, 200 rpm, for oil. Nowadays, most of the screening studies of microbial biosurfactant
9 days (Ma et al., 2012). After 144 h of cultivation this yeast was able to producers are performed using soil samples, generally from environ-
produce 41.0 g/L of a mixture of sophorolipids (acidic and lactonic ments contaminated with hydrocarbons or crude oil (Ji et al., 2016;
types), that exhibited, after purification, different profile of surface Jemil et al., 2016; Yadav et al., 2016). This perspective is widely used
activity and critical micelle concentration (CMC) values, ranging from due to the adaptive factors attributed to the microorganisms present in
32.32 mN/m up to 42 mN/m and 20 to 90 mg/L, respectively. More contaminated soils.
recently, the potential of 33 actinomycetes isolated from marine sponge Thirteen actinomycetes strains isolated from sediment samples of
Fasciospogia cavernosa were evaluated for biosurfactant production, and industrial and coal mine areas were screened by Chakraborty et al.
among them a strain identified as Nesterenkonia sp. MSA31 exhibited (2015). In this work, 4 strains showed positive results in the screening
the highest lipopeptide production capacity (Kiran et al., 2017). In this methods used (oil displacement and drop collapse methods, CTAB agar
study, all microorganisms were cultivated in minimal salt media plate method and lipase activity) and the best result was found for
(KH2PO4 0.5 g/L, FeSO4 .7H2O 0.1 g/L, Na2CO3 0.2 g/L, L-asparagine Actinomycetes nocardiopsis A17, that was able to reduce the surface
0.1 g/L, MgSO4 0.1 g/L, yeast extract 1 g/L and NaCl 1 g/L, at pH 7) and tension of water from 72 mN/m to 51.43 mN/m with CMC value around
327
6 mg/L. Keskin et al. (2015) described the production of lipopeptide and the effects of carbon and nitrogen were also investigated (Onwosi
biosurfactant by Staphylococcus xylosus STF1 isolated from petroleum and Odibo, 2012). The results suggested that glucose and sodium ni-
contaminated soil samples of Ankara, Turkey. In addition, this bio- trate are the best carbon and nitrogen sources, allowing rhamnolipid
surfactant showed antimicrobial effects against several pathogenic yields of 5.28 and 4.38 g/L, respectively. Moreover, the biosurfactant
bacteria strains, including Salmonella enterica, Sthaphylococcus aureus, produced exhibited good surface activity and critical micelle con-
Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae and Pseudomonas centration (surface tension reduction of 37 mN/m and CMC 0.028 g/L)
aeruginosa. and their application in enhanced oil recovery and petroleum de-
In a screening study performed by Jemil et al. (2016), six wild-type gradation was suggested. Previously, the production of rhamnolipids by
bacteria strains were isolated from hydrocarbon contaminated soil a strain of Rhodococcus fascians isolated from Antarctica soil samples
samples and evaluated for biosurfactant production using oil displace- was described (Gesheva et al., 2010). In addition, this strain was able to
ment and drop collapsing tests, haemolytic activity and surface tension produce biosurfactant using kerosene and glucose as carbon source,
measurement. The best results were obtained by a strain identified as which lowered the surface tension of air/water from 72 mN/m to 27
Bacillus methylotrophicus DCS1, able to reduce the surface tension of mN/m and exhibited antibacterial activity against B. subtilis ATCC
water from 72 mN/m to 31 mN/m and a CMC value of 100 mg/L. These 6633.
results were described to crude extract from acid precipitation of cell Recently, the production of rhamnolipid by an unconventional and
free supernatant and its chemical composition was reported in a recent non-pathogenic microorganism identified as Acinetobacter junii BD was
study using tandem high-resolution mass spectrometry analysis (Ma- reported by Dong et al. (2016). After a screening performed with 77
trix-assisted laser desorption ionization time of flight mass spectro- wild microorganisms isolated from oil reservoir samples, this hydro-
metry- MALDI-TOF-MS and MALDI-TOF MS2) after extraction and carbon-degrading bacteria strain was selected due to ability to produce
fractionation in silica gel column (Jemil et al., 2017). The chemical different isoforms and homologs of rhamnolipids. Moreover, the au-
analysis showed that the crude extract contained a mixture of 25 ana- thors demonstrated which using NaNO3 as nitrogen source and 10 g/L
logues of three different lipopeptides families, including isoforms of of soybean oil as carbon source the maximum concentration of biomass
surfactin, iturin and fengycin. Previously, Xia et al. (2014) also reported and biosurfactant were achieved. Rikalovic et al. (2013) performed a
the production of a mixture of isoforms and homologs of cyclic lipo- comparative analysis of rhamnolipids produced by 4 Pseudomonas aer-
peptides (surfactin, fengycin and lichenysin) using a Pseudomonas sp. uginosa isolated from hydrocarbon contaminated soil samples. In this
WJ6 strain isolated from heavy oil contaminated soil. Moreover, the study, the rhamnolipid mixtures obtained showed similar surface ac-
biosurfactant-producing bacteria was able to degrade different n-al- tivity varying from 38.8 mN/m to 40.5 mN/m, and the CMC values
kanes and polycyclic aromatic hydrocarbons and to heavy oil washing, ranging from 131 mg/L to 162 mg/L. Satisfactory results in terms of
indicating the potential use in environmental remediation and petro- production were obtained when the fermentation was carried using
leum industry. phosphate-limited proteose peptone-ammonium salt (PPAS) medium,
Recently, Joy et al. (2017) described the isolation of 47 bacterial with sunflower oil as a carbon source, obtaining the total rhamnolipid
strains, among them, 11 with potential for biosurfactant production and production ranging from 0.053 g/L up to 3.3 g/L. In this context, Ji
concomitant hydrocarbon degradation. These microorganisms were et al. (2016) reported the production of four different types of rham-
obtained from hydrocarbon contaminated soil, and extreme environ- nolipids using P. aeruginosa M408 isolated from petroleum-con-
ments samples as desert soil and hot spring water. The screening study taminated soil sample when olive oil was applied as sole carbon source
was performed in two steps: drop collapse and oil-spreading methods in the bioprocess. This strain was able to produce around 12.6 g/L after
followed by a secondary screening using five different assays, haemo- 96 h using 40 g/L of olive oil as carbon source and the mixture of
lytic assay, CTAB method, surface tension measurement, and emulsifi- rhamnolipids produced exhibited a CMC value ranged of 60–120 mg/L
cation index (E24). Four different microorganisms were reported as lowering the surface tension to values less than 31 mN/m.
producers of glycolipids (Achromobacter sp. PS1 and Bacillus sp. SLDB1) Wild strains isolated from leaves, plant tissues and fruits demon-
and lipopeptides (Ochrobactrum sp. GREW1 and Bacillus sp. SB2). All strated that important biosurfactant producers can be obtained from
the biosurfactants exhibited high emulsification activity and were ef- natural sources and in this context, a strain of Pseudozyma chura-
fective in surface tension reduction, also showing efficiently crude oil shimaensis, isolated from leaves of sugarcane (Saccharum officinarum),
degradation profile revealing their potential applicability in bior- showed the ability to produce a mixture of mannosylerythritol lipids
emediation processes. (MEL) with excellent surface properties (CMC: 0.0011 g/L and surface
Mouafi et al. (2016) reported a potent biosurfactant-producing tension reduction of 29.2 mN/m) (Morita et al., 2011a). The same type
strain identified as Bacillus brevis after screening study with 20 bacterial of biosurfactant could be produced by Ustilago scitaminea isolated from
isolates obtained of the different samples collected from oil con- sugarcane juice, being obtained 25.1 g/L of MEL (Morita et al., 2011b).
taminated soil and sediment of mangrove trees. The application of A new strain of Pseudozyma tsukubaensis isolated from leaves of Perilla
statistical tools through central composite design for optimization of frutescens was employed for diastereomer MEL-B production and ex-
biosurfactant production by B. brevis demonstrated that the best con- hibited high concentration of this biosurfactant (73.1 g/L) using a fer-
ditions for achieving high production (predicted value of E24 around mentation medium with olive oil as a carbon source in fed-batch culture
80%) was when the biotechnological process was carried with 8.5 g/L (Morita et al., 2010). This concentration of biosurfactant was obtained
of glucose as a carbon source at 33 °C and pH 8 for 10 days. Previously, after incubation at 25 °C for 168 h, in 20 mL medium containing 100 g/
the production of rhamnolipids by Serratia rubidaea SNAU02 isolated L of olive oil, and after 56 h, 100 g/L of olive oil was directly added to
from hydrocarbon-contaminated soil under solid-state fermentation the culture. In addition, these authors performed also a screening of 43
was optimized using this same statistical strategy (Nalini and fungal MEL-producers isolated from sugarcane plants. Among them, 4
Parthasarathi, 2014). The potential of mahua oil cake as substrate in strains were able to grown in culture media containing 50% sugarcane
solid-state fermentation was evaluated and the best conditions reported juice as alternative substrate, reaching biosurfactant production
were using 7.48 g/L of mahua oil cake, 2.5 mL of inoculum size, and pH varying from 0.8 to 3.7 g/L (Morita et al., 2012).
7.22 at 31 °C. According to the exposed, the isolation and screening of novel
The production of rhamnolipids biosurfactant is traditionally per- biosurfactant producers consist in an important biotechnological ap-
formed using Pseudomonas aeruginosa strains, but some studies have proach due the potential market and several applications of microbial
been reported the production of this glycolipid biosurfactant by other surfactants in medical, food, cosmetic, environmental and pharmaceu-
species, an example was the production rhmanolipids by Pseudomonas tical industries. However, more studies focused in development of cost-
nitroreducens isolated from petroleum-contaminated soil was described attractive process for biosurfactant production in large-scale are
328
necessary, as well as the standardization of the culture media and as- Kelcogel®, patented by Kelco; and pullulan, that can be produced from
says used in the screening studies. Another important topic is the use of starch by the fungus Aureobasidium pullulans and is the most used in
low-cost carbon sources and optimization strategies to improve the breath freshener or oral hygiene products, such as Listerine® (Singh
production of biosurfactants by newly microorganisms obtained from et al., 2008). Nowadays, companies such Rhone Poulenc (France),
screening studies. Finally, further efforts are expected for the devel- Statoil (Norway), and Kelco (USA) are leaders at microbial exopoly-
opment and standardization of high throughput screening methods, and saccharides research and production, with expected increase of 10%
it is suggested that the new works use the combination of traditional annually (Sidorova and Voronov, 2016).
methods for selection of potential biosurfactant producers, as well as A search in Google Patents shows that there is an increase in the
high performance analytical tools for efficient characterization of the patents about EPS since the beginning of 2000, reaching its maximum
biosurfactants produced. in the period 2013–2017. They are mostly regarding EPS producing
methods using different microbial strains and for application in food,
2.3. Polysaccharides pharmaceutical and medical fields, for example skin protection against
UV and kidney protection using EPS produced by Ceriporia lacerate; and
Polysaccharides are macromolecules belonging to the group of prevention or treatment of metabolism disorders using EPS produced by
carbohydrates, generally used as thickeners, gelling agents and stabi- Leuconostoc mesenteroides (Kim et al., 2017; Ikuo et al., 2017).
lizers in food and pharmaceutical products (Moscovici, 2015). These EPS presents an important physiological role for the microorganism,
compounds are classified considering their position inside the microbial being mainly associated with protection of the cell from adverse con-
cell, as (i) polysaccharides constituting the cell wall, such as techoid ditions, for example the formation of biofilms, responsible for resistance
acids, lipopolysaccharides (LPS) and peptidoglycans; (ii) poly- to environmental stress and disinfectants, and also for the formation of
saccharides which provide energy and act as carbon source for the cell, cell aggregates, an important characteristic for wastewater and soil
called cytosolic polysaccharides; and (iii) exopolysaccharides (EPS), treatment (Czaczyk and Myszka, 2007; Rühmann et al., 2015).
polysaccharides that are secreted, in form of biofilm or capsules, to the The amount of EPS produced is directly linked to the process con-
extracellular medium (Bergmaier, 2002; Boels et al., 2001; Canilha ditions and medium composition, and also the type of microorganism
et al., 2006; Donot et al., 2012; Lahaye, 2006; Ruas-Madiedo and de los used (Kumar et al., 2007). Factors such availability and type of carbon
Reyes-Gavilán, 2005). and nitrogen sources, temperature, pH, growth phase, agitation, and
Other classification for microbial polysaccharides are based on the aeration play the major influence, and are specific for each micro-
constitution of the molecule, being heteropolysaccharides, constituted organism (Barbosa et al., 2004; Czaczyk and Myszka, 2007). To identify
of several monosaccharides, e.g. xanthan gum and gellan; and homo- the best conditions of the different strains in order to optimize its
polysaccharides, constituted of only one type of monosaccharide, e.g. production and to better understand its metabolic ways, several studies
dextran (Fig. 4) (Aranda-Selverio et al., 2010; Bergmaier, 2002; are being conducted. In addition, the search for the appropriate mi-
Czaczyk and Myszka, 2007; Lahaye, 2006). croorganism able to accumulate polysaccharides is an emergent
The growing interest in renewable resources is leading to expansion strategy to obtain these compounds.
on the market of microbial polysaccharides, since they can be used as Table 3 summarizes the main researches dealing with screening of
substitute of polymers and they can improve the rheological char- microorganisms to produce EPS. It is interesting to notice that the
acteristics of various products in food and pharmaceutical industries methods to identify polysaccharides production in isolated strains are
(Bergmaier, 2002; Canilha et al., 2006; Donot et al., 2012; Nwodo et al., very similar in the cited articles. Most of them perform the isolation
2012; Rühmann et al., 2015; Sutherland, 1998, Sutherland, 2001). from fermented foods or from environment with harsh conditions,
The microbial polysaccharides presents itself as an unexploited where the microorganisms can produce polysaccharides as a strategy
market, specially EPS, that can be produced by various classes of mi- for cell protection (Van Geel-Schutten et al., 1998; Adebayo-Tayo and
croorganisms, such as bacteria, molds and yeasts (Sutherland, 2001). Onilude, 2008). Once isolated, the EPS-producing microorganisms are
The EPS are the most studied and important classes of microbial identified by the mucoid characteristics of their colonies. This is a fast
polysaccharides, considered potential bioactive molecules with im- and reliable method for a first screening. Then, the polysaccharide can
munostimulatory, antitumor antioxidant, antiulcer and proved reduce be better characterized and its application can be tested (Pawar et al.,
blood cholesterol. In addition, it is easier to identify the EPS-producing 2013; Subramanian et al., 2010; Vijayendra et al., 2008).
strains and also the purification of final product is simplified, reducing One of the first studies that reported the isolation of microorganisms
production costs (Madhuri and Prabhakar, 2014). For these reasons, to produce polysaccharides was conducted by Van Geel-Schutten et al.
this topic will focus only on newly isolated EPS-producting strains. (1998), where samples of fermented foods, gastro intestinal tract of
The EPS group comprises a great variety of molecules, possessing laboratory animals and human dental plaque were used. In this work,
specific properties for several industrial applications, especially at food 182 Lactobacillus sp. strains were screened for the ability to produce
and pharmaceutical fields. More than 400 EPS are known, having dif- EPS. The strains were grown anaerobically at 37 °C in liquid De Man,
ferent chemical structures and properties, depending on the different Rogosa and Sharpe (MRS) using glucose, fructose, maltose, raffinose,
sources of production (Moscovici, 2015; Rühmann et al., 2015). In this sucrose, galactose and lactose in a concentration of 100 g/L as carbon
sense, the screening and isolation of EPS-producing microorganisms sources. Sucrose was considered an excellent substrate, since only a few
and the characterization of the molecule produced represents an im- microorganisms were able to synthesize EPS from another carbon
portant step for the development of process and discovery of new in- source. Sixty strains showed positive results for EPS production, with 17
dustrial additives. strains achieving concentrations up to 100 g/L. Finally, this work in-
The first exopolysaccharide produced on industrial scale was the dicated that the higher sugar concentrations led to a large number of
xanthan gum, which continues to be one of the most commercially EPS-positive strains and a higher product yield (Van Geel-Schutten
produced nowadays (Jazini et al., 2017). The production scale reaches et al., 1998).
30,000 tons per year by the bacteria Xanthomonas campestris, ac- Kaci et al. (2005) used soil samples from eight contrasting regions of
counting for only 6% of the value of the polysaccharides market with an Algeria for the isolation of bacteria able to produce EPS. In total, 125
expected annual rate growth of 5–10% (Donot et al., 2012; Freitas et al., isolates were selected by qualitative criteria (gel-forming capacity),
2011; Habibi and Khosravi-Darani, 2017). purified and compared by EPS yields. After trials, one bacteria identi-
Other relevant EPS produced industrially are dextran, commercia- fied as a new Rhizobium sp. strain, related to R. sullae, was selected for
lized by Pharmacosmos, Sigma-Aldrich, Pharmachem Corporation and its extensive ability to produce gel-forming EPS on RCV-sucrose
Amersham Biosciences; Gellan gum, sold by the names Gelrite® and medium, reaching 2.5 g/L of EPS from 20 g/L of sucrose. The product
329
Fig. 4. Chemical structures of the most studied and commercially exploited microbial polysaccharides.
was identified and the rheological properties tested, being verified that identified using physiological and morphological tests, being all gram
it possesses a new structure, consisting of glucose, galactose, and positive, rods, cocci and ovoid, considered active producers of EPS the
mannuronic acid, which differs from others EPS produced by Rhizobium ones with yields above 40 mg/L. The results were 64.29% of
sp. and presents high viscosity stability, suggesting its use in agronomic Lactobacillus plantarum, 42.85% of L. fermentum, 50% of L. delbrueckii,
field. Leuconostoc mesenteroides, L. casei subsp. pseudoplantarum and L. lactis,
Ortega-Morales et al. (2007) screened 34 strains isolated from 100% of L. casei subsp. tolerans, L. brevis, L. coryniformis, Leuc. me-
leaves of seagrass Thalassia testudium and rock samples found in senteroides subsp. dextranicum, 40% of Leuc. gelidium, L. cellobiosus,
southern Gulf of Mexico. Samples were washed and vortexed to obtain 66.7% of Lactococcus lactis subsp. plantarum and 2.57% of Leuc. amell-
cell suspensions, diluted and incubated in marine agar (MA) at 30 °C for biosum (Adebayo-tayo and Onilude, 2008). This works shows the im-
5 days. Colonies with mucoid characteristics, indicative of EPS produ- portance of the appropriate choice of initial isolation source, since its
cers, were selected and incubated at MA + 3% glucose for 48 h. Eleven endogenous characteristics will be related with the microorganisms that
colonies were selected as good EPS-producers and from this number, 2 can be obtained. In this case, EPS gives consistency and texture in
isolates which presented anionic composition were identified as be- fermented food, therefore, the lactic acid bacteria isolates are potential
longing to the genus Microbacterium sp. and Bacillus sp.. The EPS pro- EPS producers.
duction was carried at 30 °C in 500-mL Erlenmeyer flasks filled with A similar study used dahi (a traditional fermented milk product
liquid marine broth (MB) and 30 g/L of glucose. The EPS produced by from India), buttermilk and some vegetables, such as cucumber and
Microbacterium sp. reached 2 g/L and showed an interest activity, si- cabbage, for the isolation of potential EPS-producers (Vijayendra et al.,
milar to commercial surfactants, but with higher activity of chelate 2008). Samples were inserted in sterile saline medium and subjected to
cations, being needed further studies to determine its ecological role. serial dilution plated onto modified MRS agar in which glucose was
Lactic acid bacteria were isolated from traditional fermented food in replaced for sucrose (5%, w/v) and then spread into plates and in-
MRS agar and incubated for 24 h at 35 °C. Among 113 strains isolated, cubated at 30 and 37 °C for 24–48 h, having, in total 32 colonies
101 presented potential for EPS production after trials using EPS se- identified. The colonies that presented mucoid appearance, were se-
lection medium (ESM) and incubated for 24 h at 30 °C. Product pur- lected and purified by streaking method, being identified by morpho-
ification was performed with 4 volumes of cold ethanol and quantifi- logical and physiological tests as Leuconostoc sp. The fermentation
cation by Dubois method for total carbohydrates, ranging from 1.0 to process for EPS synthesis by this isolate was conducted at 30°C, for 72 h
196.0 mg/L (Adebayo-tayo and Onilude, 2008). Microorganisms were and 200 rpm at MRS media and a new media, which facilitates the
330
M.G. Pessôa et al.
Table 3
Studies involving isolation of microorganisms for exopolysaccharides production.
Source of isolation Number of Number of positive microorganisms Process conditions Reference
microorganism
Fermented foods, gastro intestinal tract of 182 Lactobacillus strains 60 strains Anaerobically grow at 37 °C for 3 days at liquid MRS media, under Van Geel-Schutten et al.
laboratory animals and human dental agitation and flushed with nitrogen (1998)
plaque
Soil samples from Algeria 125 strains 1 strain (Rhizobium sp.) RCV-sucrose media for 5 days at 30 °C, later purified on TSA/10 Kaci et al. (2005)
medium
Leaves of seagrass Thalassia testudium and 34 strains 11 strains (Microbacterium sp. and Bacillus sp.) Marine agar (MA) at 30 °C for 5 days (Ortega-Morales et al.,
rock samples from Gulf of Mexico 2007)
Traditional fermented food from Nigeria 113 lactic acid bacteria 101 strains MRS agar, incubated for 24 h at 35 °C (Adebayo-Tayo and
strains Onilude, 2008)
Dahi, buttermilk and some vegetables, such 32 strains 1 strain (Leuconostoc sp.) Serial dilution plated onto modified De Man Rogosa and Sharpe (Vijayendra et al., 2008)
as cucumber and cabbage agar with sucrose (5%, w/v), incubated at 30 and 37 °C for
331
24–48 h
Wastewater Not mentioned 25 strains (Bacillus sp., Serratia sp., Pseudomonas sp., Enterobacter Plate count agar (PCA) and Sabouraud’s dextrose agar (SDA) (Subramanian et al.,
sp., Yersinia sp., Microbacterium sp., Pantoea sp., Photorhabdus sp. media with serial dilution incubated for 48–72 h at 30 °C 2010)
and Pectobacterium sp.)
Saline soil of Baramati region of Maharashtra Not mentioned 5 strains (Azotobacter sp., Pseudomonas sp., Agrobacterium sp., Serial dilutions and 0.1 mL of soil suspensions spread on nutrient Pawar et al. (2013)
(India) Alpha Proteobacterium group and Xanthomonas sp.,) agar plates, incubated at 48 h at room temperature
Raw camel milk from Algeria 82 strains 30 strains (12 Lactobacillus sp., 9 Streptococcus sp., 7 Enterococcus Isolation using standard pour-plate method with modified (Abdellah et al., 2014)
sp. and 2 Pediococcus sp.) Chalmers-agar media. Plates incubated in a semi anaerobically
condition for 48–72 h at 42 °C.
Environmental samples, such as soil, curd, Not mentioned 11 strains Serial dilution and incubation at 30 °C for 24 h in a medium Rawal et al. (2016)
spoilage fruit and garbage water composed of glucose (20 g/L), yeast extract (3.0 g/L), MgSO4
(0.2 g/L), K2HPO4 (5 g/L) and agar (30 g/L)
Madeira archipelago ocean sediments 744 strains 500 strains (Among them, Pseudoalteromonas sp., Alteromonas sp., Not mentioned Roca et al. (2016)
Psychrobacter sp., and Brevibacterium sp)
Kefir milk 22 strains 10 strains (Lactobacillus kefiranofaciens, Lactobacillus kefiri, A loop of kefir milk streaked onto MRS agar and incubated Jeong et al. (2017)
Lactococcus lactis, and Leuconostoc mesenteroide) anaerobically at 30 °C for 72–96 h
recovery processes. This new medium had beef extract and protease For the isolation of lactic acid bacteria, a loop of kefir milk was
peptone and presented a higher amount of EPS produced (22.5 g/L) streaked onto MRS agar and the plates were incubated anaerobically at
when compared to MRS (14 g/L), indicating that the production of EPS 30 °C for 72–96 h. By this methodology, it was possible to obtain 22
is facilitated using rich nutrients. strains, being 8 strains of Lactobacillus kefiranofaciens, 2 strains of
Subramanian et al. (2010) isolated microorganisms from waste- Lactobacillus kefiri, 7 strains of Lactococcus lactis, and 5 strains of
water in Quebec, Canada. The isolation technique used was serial di- Leuconostoc mesenteroides. Among these, 10 strains showed potential for
lution spread in plate count agar (PCA) and Sabouraud’s dextrose agar EPS production, confirming the high ability of lactic acid bacteria to
(SDA) media and incubated for 48–72 h at 30 °C. From the 25 bacterial synthetized EPS. The Lactobacillus kefiranofaciens was identified as the
strains with mucoid appearance, 13 of them were gram positive. Using best producer strain with highest yield of EPS, 2.2 g/L. This EPS showed
16s rDNA sequence, 11 bacterial strains were identified as Bacillus sp., 3 antimicrobial activity against Listeria monocytogenes and Salmonella
Serratia sp., 2 Pseudomonas sp., 2 Enterobacter sp., 2 Yersinia sp., 2 Mi- Enteritidis suggesting a novel bioactive compound in kefir (Jeong et al.,
crobacterium sp. and 1 strain from each genus Pantoea, Photorhabdus and 2017).
Pectobacterium, producing EPS with concentrations ranging from 3.6 to It is noteworthy that due to the great diversity of polysaccharide-
35.8 g/L. producing microorganisms and the different uses and properties of
Five strains of bacteria were isolated from saline soil of Baramati these compounds, the search for strains capable of synthesizing poly-
region of Maharashtra (India) and identified by their morphological, saccharides through isolation and screening represents a useful bio-
biochemical characteristics as well as 16S rRNA gene sequencing, as technological tool, able to discover new strains and new poly-
Azotobacter sp., Pseudomonas sp., Agrobacterium sp., Alpha saccharides structures. Even though there are some successful cases of
Proteobacterium group and Xanthomonas sp., with ability to produce EPS. screening procedures with basic conditions and many isolated micro-
The choice of saline soil was based on the fact that this condition could organism presenting satisfactory amounts of EPS produced, such as
generate stress at the natural microflora. The screening of the bacteria described by Jeong et al. (2017), Roca et al. (2016), Van Geel-Schutten
was made with serial dilutions and 0.1 mL of soil suspensions spread on et al. (1998) and others cited above, further studies with the newly
nutrient agar plates and incubated at 48 h at room temperature. The isolated strains are necessary to determine the most influential condi-
mucoid colonies were screened and re-streaked on another nutrient tions and optimized values, in order to obtain maximum production and
agar plate to obtain pure culture. The EPS production by the strain yields.
Agrobacterium sp. achieved best results, having its production condi-
tions optimized obtaining 5.2 g/L at pH 7.5, 30 °C and 3 days of fer- 2.4. Microbial oils
mentation (Pawar et al., 2013).
Abdellah et al. (2014) isolated thermophilic lactic acid bacteria for Microbial oils, also known as single cell oils (SCOs), have being
the production of EPS from raw camel milk in Algeria. In total, 28 broadly studied as an alternative for the production of conventional
samples of camel milk were taken from two different locations at south oils, mainly due to their fatty acids profile similarity with vegetal oils.
Algeria and acidified to retrieve a large amount of lactic acid bacteria. Therefore, these compounds could be a viable source for the synthesis
The procedure consist of 10 ml of each sample mix with 90 ml of sterile of cocoa butter substitute, feedstock for biodiesel production or as
yeast water (10% w/v) followed by serial decimal dilutions. Then, the source of polyunsaturated fatty acids, like a γ–linoleic acid (GLA)
isolation of the bacteria was made using the standard pour-plate (Fig. 5) (Arous et al., 2015; Vieira et al., 2014; Bharathiraja et al.,
method with modified Chalmers-agar medium. After, the plates were 2017). Besides that, SCOs can also be applied in the production of
incubated in a semi anaerobic condition for 48–72 h at 42 °C. The co- biosurfactants, solvents, paints, plastics, lubricants and waxes (Ayadi
lonies were purified on MRS agar plates, being found 82 different et al., 2016; Vinarta et al., 2016). Additionally, these products have
strains, of these, 30 strains presented potential for produce EPS (12 great appeal for use in the food industry, as recently reviewed by
Lactobacillus sp., 9 Streptococcus sp., 7 Enterococcus sp. and 2 Pediococcus Béligon et al. (2016).
sp.), with concentrations range from 70 to 740 mg/L, being necessary SCOs can been produced by oleaginous microorganisms such as
more studies for identification of EPS and properly utilization (Abdellah fungi, yeasts, bacteria and microalgae (Subhash and Mohan, 2011).
et al., 2014). Nevertheless, to be considered as a producer of such compounds, mi-
More recently, Rawal et al. (2016) isolated microorganisms from croorganisms should accumulate at least 20% of their dry biomass as
environmental samples, such as soil, curd, spoilage fruit and garbage lipids (Ayadi et al., 2016; Cui et al., 2012) and this capacity is generally
water using serial dilution and incubation at 30 °C for 24 h in a medium analyzed by Nile Red Fluorescence or Sudan Black B staining. Ap-
composed of glucose (20 g/L), yeast extract (3.0 g/L), MgSO4 (0.2 g/L), proximately 80–90% of microbial oils are triglycerides containing long
K2HPO4 (5 g/L) and agar (30 g/L). Colonies with mucoid appearance chain fatty acids (Ayadi et al., 2016; Vinarta et al., 2016). This oils
(eleven, in total) were obtained at pure form at agar plates. The pro- could be considered similar to vegetable oils when their fatty acid
duction of EPS was conducted by submerged fermentation at 30 °C, profile have abundant quantities of low degree unsaturated long chain
150 rpm for 24 h. The maximum EPS production was obtained after fatty acid (as oleic and linoleic acids) and saturated long chain fatty
24 h of fermentation with a yield of 3.65 g/L. One newly isolated strain acids (as palmitic and stearic acids) (Ayadi et al., 2016; Liu et al., 2010;
called P11 had its EPS production optimized by one-factor-at-a-time Tampitak et al., 2015; Vinarta et al., 2016). This fatty acid profile can
with various nitrogen sources, carbon source, incubation temperature, be analyzed by GC-MS after a transesterification process (Khot et al.,
pH analyzed, with the best operational conditions at pH 7, 30 °C, 4.5 g/ 2012; Liu et al., 2010).
L of glucose and 5.9 g/L of beef extract. Is noteworthy that the fatty acids profile produced and its quantity
From 744 microorganisms isolated from Madeira archipelago ocean depend fundamentally on the catalyst, carbon source, pH and tem-
sediments, Roca et al. (2016) selected 500 strains as potential EPS perature of the medium, demonstrating the crucial role of process
producers being that only 8% presented yields above 1.0 g/L and 2% conditions in the production of lipids of economic interest, as can be
presented EPS concentration above 2.5 g/L. The strains were tested at seen in the current text, the working ranges are in the most cases at a
25 mL shake flasks on liquid medium (750 mL sea water and 250 mL pH between 5.5–6.5, temperature 28–30 °C and agitation of
deionized water) containing: 30 g/L glucose, 4 g/L yeast extract and 150–250 rpm (Khot et al., 2012; Donot et al., 2014). Based on these
2 g/L peptone, incubated for 5 days at 28 °C and 200 rpm. Among them, limitations, several strategies were conducted in order to increase
Pseudoalteromonas sp., Alteromonas sp., Psychrobacter sp., and Brevi- biomass productivity and lipid content, as well as favor the production
bacterium sp. were identified, being Pseudoalteromonas sp. the best of a compatible fatty acid profile for the application as raw material in
producer, with yield of 4.4 g/L after 17 h of fermentation. biodiesel production. These strategies include the isolation of
332
Fig. 5. Fatty acids found in microbial oils produced by fermentation.
microorganisms from hostile environments, use of raw material as samples from different regions of Thailand. Among these, 48 strains
carbon source, scale up process and process optimization (Kraisintu presented potential as microbial oils producer. The best producer,
et al., 2010; Saran et al., 2017). classified as Rhodosporidium toruloides strain DMKU3-TK16 was used in
Polyunsaturated fatty acids have a crucial role in maintaining bio- a study for optimizing process parameters; reaching a lipid concentra-
functions in humans (Ratledge, 2013), and due to their low availability tion of 9.26 g/L in a medium containing 70 g/L of glucose and pH of 5.5
in animal and plant origin foods, they are sometimes consumed as and at 28 °C.
nutraceutical (Finco et al., 2016). Some of these compounds, such as Aiming the isolation of oleaginous Mucor sp. strains with potential
docosahexaenoic acid (DHASCO®), eicosapentaenoic acid (EPA-SCO), for the synthesis of γ–linoleic acid (GLA), Mamatha et al. (2010) col-
GLA (Oil of Javanicus) and arachidonic acid (CABIO oil and ARA-SCO), lected 250 soil samples from different regions, reaching up 20 species.
are currently produced commercially by different type’s microorgan- From these, 9 showed satisfactory results for the production of lipids
isms, however the production of biodiesel from microbial oil is not reaching lipid concentrations ranging between 20.08 and 30.0%. The
viable in the current market (Finco et al., 2016; Ochsenreither et al., fungus Mucor rouxii identified as CFR-G15, reached 2.97 g/L of lipids
2016; Sitepu et al., 2014; Ratledge, 2013; Huang et al., 2013; Jin et al., produced, of which 14.42% were GLA, when was used glucose as
2015). Also many patents have been recently registered to describe the carbon source (40 g/L), at 28 °C and agitation of 200 rpm. This strain
process production of microbial oil with genetic modified strains, pro- appears to be a potential candidate for the production of GLA.
duction of microbial oil with high proportion of polyunsaturated fatty Out of 61 microorganisms isolated from forest soils in Hunan pro-
acids (PUFA), new process to isolation microbial oil without organic vince in China and cultured in high glucose (30 g/L) medium, tem-
solvent and the use of lignocellulosic materials as substrates perature of 30 °C, 180 rpm and pH of 5.5, 16 strains showed positive
(Stephanopoulos et al., 2013; Trimbur et al., 2016; Trimbur et al., 2008; profile for the production of lipids. The strain identified as Thamnidium
Hong et al., 2012; Yamaguchi et al., 2016; Biji and Schaap, 2004; ctenidium presented the best concentration of lipids (6.4 g/L). This mi-
Koskinen and Tanner, 2012; Vainio et al., 2016; Yu et al., 2014). croorganism was then cultured in a 5-L bioreactor under optimal con-
The production of SCOs is a recent and extensively explored topic, ditions of glucose (60 g/L), pH (6.5), temperature (30 °C) and agitation
since most of the microbial oils features as interesting fatty acid profile (220 rpm), yielding 20.6 g/L of biomass and 13.6 g/L of lipids, which
and similar to vegetable oils, increasing their potential for biodiesel showed fatty acid profile similar to vegetable oils (Liu et al., 2010).
production (Tanimura et al., 2014). Biodiesel is produced by transes- A culture of Aspergillus sp. was isolated from marshy soil samples
terification of ethanol or methanol with vegetable oils, which are also and grown on glucose or corncob waste liquor (CWL) as carbon sources.
intended for human consumption (Liu et al., 2010; Vieira et al., 2014). The best results were obtained under pH of 5.0, 150 rpm and 30 °C,
Associated with this, biodiesel produced by this method gives a product using glucose as substrate, reaching 23.33% lipids, whereas the results
of higher cost than conventional diesel, causing an obstacle to its use for the CWL were 22.1% lipids (Subhash and Mohan, 2011).
(Pan et al., 2009). Considering the potential of such products, the aim of Eleven microorganisms were isolated from soil samples collected
this section is to present the main microorganisms screened for the from hot spring, wetland, sand, lawn, saline and alkali land, 7 of which
production of microbial oils, which can be used for various purposes were capable of producing lipids in significant amounts (more than
but mainly as raw material the production of biodiesel. 20%) using D-xylose as carbon source. Among these, the three strains
Pan et al. (2009) isolated 40 yeast colonies from orchard soils, from identified as Tilletiopsis albescens, Aspergillus fumigatus and Gibberella
which 20 of them showed potential to produce lipids when cultivated in fujikuroi produced lipids ranging from 3.74 to 4.93 g/L, with similar
a medium containing xylose (40 g/L) as carbon source, at 28 °C and composition to that of vegetable oils (Li et al., 2011).
agitation of 180 rpm. Among them, 13 colonies showed lipids pro- Kitcha and Cheirsilp (2011) isolated yeast strains from soils and
ductivity at satisfactory levels (higher than 2.5 g/L), which indicates wastes of palm oil mill and biodiesel plant using glucose or glycerol as a
their potential for biodiesel production or as substitute for coconut oil. carbon source (40 g/L). From 889 isolated strains, only 23 presented
Among 83 yeast strains isolated from soil samples from Ermei positive results for the production of lipids. These strains were then
(China), 16 showed potential for the production of oils and 12 were incubated with crude glycerol as carbon source and four of them pre-
capable of development using xylose (50 g/L) as carbon source. The sented lipid content higher than 40%. Under optimal process conditions
highest lipid content found was 51.43% by one xylose negative strain with crude glycerol of 10%, C/N ratio of 17 and 0.5% ammonium
when cultivated with glucose (50 g/L) as carbon source. However, the phosphate, the most productive strains were Trichosporonoides spathu-
yeast identified as Pichia guillermondii stood out as the best microbial oil lata and Kodamaea ohmeri, achieving concentration of lipids equal to
producer, reaching approximately 6.2 g/L of product (He et al., 2010). 4.45 g/L and 3.22 g/L, respectively.
Kraisintu et al. (2010) isolated more than 100 strains from soil In a later study, the same authors performed the scale-up process in
333
a 5-L bioreactor, using Trichosporonoides spathulata as biocatalyst and that this yeast reproduced both sexually and asexually, depending on
crude glycerol as carbon source (10% w/v). Authors compared batch the supply of nutrients (C/N ratio), resulting in greater quantities of
and fed batch system for process improvements, where cells production biomass when has asexual reproduction (7 g/L) and a reduction of
reached 11.3 g/L and 17.3 g/L while lipid content equal to 44.3 and biomass and increased concentration of lipids from 11.9 to 22.4% when
41.9%, respectively, demonstrating the importance of adapting a sui- passed to the sexual reproductive stage (Arous et al., 2015).
table process to maximize lipid production (Kitcha and Cheirsilp, Polburee et al. (2015) isolated 323 yeasts from several samples of
2013). soil and other materials and another 64 yeast strains from plant leaves.
Khot et al. (2012) isolated 14 microorganisms from mangrove Among this total, 23 species accumulated lipids in concentrations
wetlands along India west coast, including five filamentous fungi that higher than 20% of dry biomass when cultivated at 28 °C, pH 5.5 and
were able to accumulate lipids. The fungus identified as Aspergillus agitation of 150 rpm. When pure glycerol was used as substrate, the
terreus Thom-309 showed a higher percentage of lipids (54%), with highest yields were achieved by the strain Rhodotorula taiwanensis
maximum lipid accumulation of 2.98 g/L. The fatty acid profile of the DMKU-RK65, that reached 7 g/L (71% lipids), while Rhodosporidium
lipid produced proved to be very similar to conventional vegetable oils fluviale DMKU-RK253 using crude glycerol, accumulated 3.9 g/L of li-
and presented specifications inside the international standards for pids.
biodiesel production, such as density, kinematic viscosity and con- Among 138 yeast strains isolated from soil and wastes of palm oil
centrations of linolenic acid and fatty acid methyl esters (FAME). Pro- mill, 16 were able to produce lipids from glucose (20 g/L) and 11 when
cess conditions that led to higher biomass and lipid production were xylose (20 g/L) was used as carbon source at 30 °C, 140 rpm of agitation
30 °C, 120 rpm and pH 5.5. and pH 6.0. However, only 3 of these strains showed positive result for
Cui et al. (2012) isolated 55 pure colonies from salt marsh en- the production of lipids, when used EFB hemicellulose hydrolysate
vironment, being first grown in liquid medium containing sea salt and (20 g/L) as carbon source. These strains were identified as Rhodotorula
using glucose as carbon source. However, only the psychrotrophic mucilaginous G43, Kluyveromyces marxianus X32 and Candida tropicalis
strain Sporobolomyces roseus showed satisfactory results, with a pro- X37. This work demonstrated that maximum lipid concentration ob-
duction of approximately 2.37 g/L of total fatty acids, of which 1.86 g/L tained was approximately 2.73 g/L and could be used as feedstock for
were unsaturated fatty acids. biodiesel production considering the fatty acid composition similar to
A Brazilian group isolated 129 strains of yeasts from soil samples, than those of vegetable oils (Tampitak et al., 2015).
stems, fruits and flowers, collected from four Brazilian biomes. From From samples collected in Antarctica, it was isolated 12 oleaginous
these strains, 42 showed potential for the production of lipids, but only yeast strains that were able to accumulate lipids between 20.4 and 73%
5 were evaluated for productivity of biomass and lipids. Tests were m/m of dry biomass. Among them, the strain identified as Rhodotorula
conducted in laboratory scale (Erlenmeyer flasks), under pH 6.0, 28 °C glacialis R15 showed the best results, accumulating 73% m/m of lipids.
and 185 rpm of agitation. One isolated strain from the Pantanal and It was observed a high amount of linolenic acid in this microbial oil,
classified as Candida sp. LEB-M3 showed lipids concentrations of and despite its possible application as feedstock for production of bio-
2.43 g/L and 9.14 g/L, grown with pure and raw glycerol, respectively. diesel, this composition also indicates a potential application in the
Fatty acids profile obtained from raw glycerol proved to be very similar food and pharmaceutical industries. Additionally, this work also iden-
to soybean oil, demonstrating its potential application for the produc- tified a new strain of oleaginous yeast called Rhodotorula laryngis. All
tion of biodiesel (Duarte et al., 2013). microorganisms were cultured with glucose 30 g/L as carbon source,
A Pichia kudriavzevii strain was selected as highly efficient lipid- pH 6.0, 250 rpm and 25 °C for psychrotolerant or 15 °C for psychro-
producer from 250 microorganisms isolated from samples of several philic strains (Vinarta et al., 2016).
fruits. Optimization of process parameters showed that the best Ayadi et al. (2016) tested the lipid accumulation capacity of the
growing conditions was the use of glycerol (1% v/v) as carbon source, yeast Candida viswanathii Y-E4, isolated from soil samples extensively
pH 5.5, 200 rpm agitation and 28 °C (Sankh et al., 2013). Following, contaminated by lipid substances. This yeast was grown in Erlenmeyer
bioprocess scale-up was conducted in fed-batch system in 5-L and 26-L flasks and incubated with 40 g/L of glucose as carbon source, achieving
bioreactors. Authors used crude glycerol (10 g/L) as the carbon source, concentrations of 3.2 g/L of lipids (Ayadi et al., 2016). Subsequently,
700 rpm, pH 5.5 and 28 °C, reaching cell concentration of 35.7 g/L and authors noted through carbon source optimization that the best yields
32.1 g/L, while lipids concentration reached 18.6% and 16.6%, re- were achieved using oleic acid, when process reached 18.17 g/L of
spectively. Fatty acid profile showed that this product could be a sui- biomass and 11.81 g/L of lipids. Fatty acids profile obtained was found
table source for biodiesel production due to their similarity with other to be very similar to the profile of fatty acids of vegetable oils and
microbial oils produced by Cryptococcus curvatus (Sankh et al., 2013). therefore a potential candidate for the production of biodiesel. Finally,
Tanimura et al. (2014) selected 13 lipid-producer strains among 500 the ability of Candida viswanathii Y-E4 of producing lipids using agro-
isolated yeast strains. Cryptococcus sp. strain JCM 24511 showed higher industrial wastes as alternative substrate was evaluated. Interesting
lipid content among the tested strains, reaching 1.37 g/L (approxi- results were achieved using soap stock of soybean oil refining, reaching
mately 61.5%) and 2.23 g/L of biomass using glucose as carbon source, 5.93 g/L of lipid content, whereas the higher concentration of lipid
using 150 rpm at 27 °C. (45.42%) were achieved using waste cooking oil (Ayadi et al., 2016).
Kitcha and Cheirsilp (2014) isolated 51 microorganisms from soils In a recent study, 139 yeast strains were isolated from soil samples
and wastes of palm oil mills. Only 4 were selected for lipid production rich in cellulosic wastes. Among them, 23 were identified as oleaginous
studies in solid-state fermentation (SSF), due to their potential pro- yeasts when cultivated with glucose (40 g/L) as carbon source. The
duction of lipids and high cellulolytic activity. The fungus identified as strain identified as Cystobasidium oligophagum JRC1 presented the
Aspergillus tubingensis TSIP9 were able to synthesize lipids using palm highest levels of lipids by Nile Red fluorescence microscopy protocol,
pressed fiber (PPP) and palm empty fruit bunch (EFB) as a carbon being also cultivated with different carbon sources (40 g/L), such as
sources in the proportion of 31.1 and 37.5 mg/g of dry substrate, re- starch, CMC, glycerol, xylose, maltose, fructose, sucrose, lactose and
spectively, and after optimization process the fungal strain was able to glucose. The best results obtained by C. oligophagum JRC1 in the carbon
produce 88.5 mg/g of lipid. The profile of the lipid produced demon- source test for biomass production were glucose (12.45 g/L) and fruc-
strated that this product could be used as an interesting source for the tose (11.5 g/L); while for lipid output was glucose (4.91 g/L) and xylose
production of biodiesel(Kitcha and Cheirsilp, 2014). (4.02 g/L) and for lipid content was glycerol (42.7%) and starch
The yeast identified as Debaryomyces etchellsii, isolated from a (41.54%) (Vyas and Chhabra, 2017).
sample of olive mill wastewater sludge, was cultivated in 3.7-L bior- Saran et al. (2017) isolated 52 yeast strains from decaying peel of
eactor batch system with glucose as carbon source. Authors reported banana, mango, litchi, grapes, melon, and plum, being that 4 strains
334
Table 4
Main microorganisms isolated from different environments, substrates used and achieved yields for microbial oils.
Microorganism Isolation source Substrate Biomass (g/L) Lipids (g/L) Reference
Aspergillus sp. Marshy soil samples Glucose 13.6 g/L 3.17 g/L Subhash and Mohan (2011)
Sporobolomyces roseus Salt marsh Glucose and sea salts 7 g/L 2.37 g/L Cui et al. (2012)
Rhodosporidium toruloides DMKU3- Soil samples of Thailand Glucose (70 g/L) 13.33 g/L 9.26 g/L (Kraisintu et al., 2010)
TK16
Aspergillus terreus Thom 309 Samples from mangrove Glucose (30 g/L) 5.52 g/L 2.98 g/L Khot et al. (2012)
wetlands
Mucor rouxii CFR-G15 Soil samples Glucose (40 g/L) 8.8 g/L 2.97 g/L Mamatha et al. (2010)
Cryptococcus sp. Samples from Kyoto University Glucose 2.23 g/L 1.37 g/L Tanimura et al. (2014)
Debaryomyces etchellsii Olive mill wastewater sludge Glucose - 22,4% Arous et al. (2015)
Candida viswanathii Y-E4 Soil samples Soap stock of soybean oil 14.48 g/L 5.93 g/L Ayadi et al. (2016)
refining
Glucose 21 g/L 10.5 g/L
Rhodotorula glacialis R15 Samples collected in Antarctica Glucose (30 g/L) – 6.92 g/L Vinarta et al. (2016)
Thamnidium ctenidium Forest soil samples Glucose (60 g/L) 20.6 g/L 13.6 g/L Liu et al. (2010)
Cystobasidium oligophagum JRC1 Soil samples Glucose (40 g/L) 12.45 g/L 4.91 g/L Vyas and Chhabra (2017)
Rhodosporidium toruloides A29 Decaying peel of fruits Glucose (75 g/L) 23.36 g/L 12.5 g/L Saran et al. (2017)
Rhodococcus sp. YHY01 Water samples Glucose (3% w/v) 3.27 g/L 1.54 g/L Bhatia et al. (2017)
Candida sp. LEB-M3 Samples of Pantanal Raw glycerol 16.12 g/L 2.43 g/L Duarte et al. (2013)
Candida sp. LEB-M3 Samples of Pantanal Pure glycerol 11.86 g/L 9.14 g/L
Rhodotorula taiwanensis DMKU- Soil samples and other Pure glycerol (70 g/L) 9.8 g/L 7 g/L Polburee et al. (2015)
RK65 materials
Rhodosporidium fluviale DMKU- Soil samples and other Crude glycerol 6.0 g/L 3.9 g/L
RK253 materials
Pichia kudriavzevii Fruits samples Crude glycerol (10 g/L) 35.7 g/L 6.64 g/L Sankh et al. (2013)
Trichosporonoides spathulata Soils and wastes of palm oil mill Crude glycerol (10 w/v%) 10.40 g/L 4.45 g/L Kitcha and Cheirsilp (2011)
Kodamaea ohmeri and biodiesel plant 10.50 g/L 3.22 g/L
Trichosporonoides spathulata 11.3 g/L 5.01 g/L Kitcha and Cheirsilp (2013) and
Trichosporonoides spathulata 17.3 g/L 7.25 g/L Kitcha and Cheirsilp (2013)
13 yeast Orchard soils Xylose (40 g/L) – Higher than Pan et al. (2009)
2.5 g/L
Pichia guillermondii Soil samples from Ermei Xylose (50 g/L) 13.6 g/L 6.2 g/L He et al. (2010)
(China)
Tilletiopsis albescens Soil samples Xylose (100 g/L) 15.38 g/L 4.28 g/L Li et al. (2011)
Aspergillus fumigatus 15.27 g/L 4.93 g/L
Gibberella fujikuroi 16.45 g/L 3.74 g/L
Trichosporon sp. RW Decayed wood Acid hydrolysate of sugarcane 25.28 g/L 10.25 g/L Brar et al. (2017)
bagasse
Glucose 35.98 g/L 21.45 g/L
Aspergillus tubingensis TSIP9 Soils and wastes of palm oil mill Palm empty fruit bunches and – 88.5 mg/g of Kitcha and Cheirsilp (2014)
and biodiesel plant palm kernel cake substrate
Candida tropicalis X37 Soil and wastes of palm oil mill EFB hemicellulose hydrolysate 6.81 g/L 2.73 g/L Tampitak et al. (2015)
(20 g/L)
Cryptococcus psychrotolerans River samples Groundnut shell hydrolysate 13.68 g/L 6.34 g/L Deeba et al. (2017)
IITRFD
Rhodosporidium toruloides DEBB Data not shown Sugarcane-based medium 40.3 g/L 20.5 g/L Soccol et al. (2017)
5533
were identified as lipid producers by Nile Red fluorescence. Among acid profile has indicated suitability for biodiesel production (Brar
them, the highest potential for lipid production was observed using the et al., 2017).
newly isolated Rhodosporidium toruloides A29, which was still selected Cryptococcus psychrotolerans IITRFD was selected among 386 strains
for further trials. The scale-up process was conducted in fed batch isolated from river samples (West Bengal – India). The strain showed
system in a 30-L bioreactor with glucose (75 g/L) as carbon source, pH lipid productivity of 0.044 g/L/h (6,34 g/L) when was cultivated in
5.5, 30 °C, 250 rpm, 0.6 vvm for 60 h. The scale-up provided several groundnut shell hydrolysate (GSH) as growth media at 25 °C during
improvements to the process, resulting in 12.5 g/L of yeast oil and 144 h (Deeba et al., 2017).
53.5% of lipid yield against only 14.1% reached during the screening The strain Rhodosporidium toruloides DEBB 5533 was screened
procedure (Saran et al., 2017). among one hundred different strains for microbial oil production and
The potential of lipid production of six different bacteria isolated used as biocatalyst in a pilot scale biodiesel production. The strain was
from water samples was evaluated by Nile Red and Sudan Black B cultivated in a sugarcane-based medium with urea as nitrogen source
staining and the strain identified as Rhodococcus sp. YHY01 presented during 24 h (batch) or 48 h (fed-batch) at 32 °C in a 1500 L bioreactor.
the best results. Later, this strain was cultivated with glucose or xylose The lipid production was 10.1 g/L and 20.5 g/L for the batch and fed
(1–5%) as carbon source, the maximum fatty acid production (1.54 g/L) batch system, respectively. The economic analysis was estimated the
and fatty acid accumulation (47%) were obtained at 3% glucose (Bhatia final cost of the microbial biodiesel as US$0.76/L, similar to the im-
et al., 2017). ported fossil diesel (US$0.80/L) and biodiesel from soybean (US$0.81/
A strain isolated from decayed wood, Trichosporon sp. RW, was L). Also, this microbial biodiesel has a better performance in the engine
evaluated for lipid production from glucose, glycerol or acid hydro- tests and lower pollutant emissions (Soccol et al., 2017).
lysate of sugarcane bagasse as substrate at 30 °C during 120 h of fer- Table 4 summarizes the main microbial lipid-producers selected in
mentation. This strain accumulated 21.45 (59.6%), 18.41 (56%) and screening processes, as well as their source of isolation, carbon source
10.25 g/L (40.5%) of lipids in the presence of glucose, glycerol and acid and productivities of biomass and lipids achieved. The use of microbial
hydrolysate of sugarcane bagasse, respectively. The analysis of the fatty lipids as dietary supplements can be considered as commercial reality.
335
Meanwhile, their use as starting raw materials for biodiesel production 901–914.
still lacks economic competitiveness due to the low-cost vegetable oil Balan, S.S., Kumar, C.G., Jayalaskshmi, S., 2016. Pontifactin, a new lipopeptide bio-
surfactant produced by a marine Pontibacter korlensis strain SBK-47: purification,
substrates and petroleum derivatives. Thus, the cost reduction asso- characterization and its biological evaluation. Process Biochem. 51, 2198–2207.
ciated with better production process become important to make fea- Balan, S.S., Kumar, C.G., Jayalaskshmi, S., 2017. Aneurinifactin, a new lipopeptide bio-
sible the industrial production os such compounds. In this perspective, surfactant produced by a marine Aneurinibacillus aneurinilyticus SBP-11 isolated from
Gulf of Mannar: purification, characterization and its biological evaluation.
the combination between screening of potential strains, the use of low- Microbiol. Res. 194, 1–9.
cost substrates and process optimization should improve the biopro- Banat, I.M., Satpute, S.K., Cameotra, S.S., Patil, R., Nyayanit, N.V., 2014a. Cost effective
duction of new compounds with commercial interest, such as the re- technologies and renewable substrates for biosurfactants production. Front.
Microbiol. 5, 1–18.
cently microbial biodiesel produced in pilot scale by Soccol et al. Banat, I.M., Díaz De Rienzo, M.A., Quinn, G.A., 2014b. Microbial biofilms: biosurfactants
(2017), with approximate cost around US$ 0.76/L. as antibiofilm agents. Appl. Microbiol. Biotechnol. 98, 9915–9929.
The importance of the selection of new microorganisms and po- Barbosa, A.M., Cunha, P.D.T., Pigatto, M.M., Corradi, L., 2004. Produção e Aplicações de
Exopolissacarídeos Fúngicos. Semina: Ciências Exatas E Tecnológicas 25 (1), 29–41.
tential microbial-producers is evident for this area, which constitutes a
Béligon, V., Christophe, G., Fontanille, P., Larroche, C., 2016. Microbial lipids as potential
key tool in the improvement of biosynthesis of these important com- source to food supplements. Curr. Opin. Food Sci. 7, 35–42.
pounds, since these new biocatalysts are able to produce microbial li- Berger, R., 2007. Flavours and Fragrances: Chemistry, Bioprocessing and Sustainability.
pids in sufficient quantity and quality to meet a variety of industrial Chemistry and Materials Science. Springer-Verlag, Berlin.
Berger, R., 2015. Biotechnology as a source of natural volatile flavours. Curr. Opin. Food
applications. Sci. 1, 38–43.
Bergmaier, D., 2002. Production d’exopolysaccharides par fermentation avec des cellules
3. Conclusion immobilisées de LB. Rhamnosus RW-9595M d’un milieu à base de perméat de
lactosérum. Resume 2, 2–3. Available at. http://archimede.bibl.ulaval.ca/
archimede/fichiers/20383/20383.pdf.
Considering the extent of application of biotechnological in- Bertrand, S., Bohni, N., Schnee, S., Schumpp, O., Gindro, K., Wolfender, J.L., 2014.
gredients and the essential role of microorganisms in these processes, Metabolite induction via microorganism co-culture: a potential way to enhance
chemical diversity for drug discovery. Biotechnol. Adv. 32, 1180.
there is a great interest in the selection of new biocatalysts able to resist Bhangale, A., Wadekar, S., Kale, S., Bhowmick, D., Pratap, A., 2014. Production of so-
the conditions of industrial processes, with high selectivity and high phorolipids synthesized on castor oil with glucose and glycerol by using Starmerella
productivity rates, besides being able to generate important compounds bombicola (ATCC 22214). Eur. J. Lipid Sci. Technol. 116, 336–343.
Bharathiraja, B., Sridharan, S., Sowmya, V., Yuvaraj, D., Praveenkumar, R., 2017.
with commercial interest, enabling the association of the production Microbial oil – a plausible alternate resource for food and fuel application. Bioresour.
scale with an affordable cost of biotechnological products. Technol. 233, 423–432. https://doi.org/10.1016/j.biortech.2017.03.006.
In this sense, isolation and screening of novel biosurfactant, bio- Bhatia, S.K., Kim, J., Song, H.-S., Kim, H.J., Jeon, J.-M., Sathiyanarayanan, G., Yoon, J.-J.,
Park, K., Kim, Y.-G., Yang, Y.-H., 2017. Microbial biodiesel production from oil palm
flavors, microbial lipids and polysaccharides producers, consist in an
biomass hydrolysate using marine Rhodococcus sp. YHY01. Bioresour. Technol. 233,
important biotechnological approach due to the several applications of 99–109.
these compounds in food, medical, cosmetic, environmental and phar- Bicas, J.L., Pastore, G.M., 2007. Isolation and screening of D-limonene-resistant micro-
maceutical industries. Thus, the screening and isolation of microorganisms. Braz. J. Microbiol. 38, 563–567.
Bicas, J.L., Dionísio, A.P., Pastore, G.M., 2009. Bio-oxidation of terpenes: an approach for
organisms represents an important step for the development of bio- the flavor industry. Chem. Rev. 109, 4518–4531.
processes and discovery of new products in quantity and quality to meet Bicas, J.L., Silva, C., Dionísio, A.P., Pastore, G.M., 2010. Biotechnological production of
a variety of industrial applications. bioflavors and functional sugars. Ciênc. Tecnol. Aliment. 30, 7–18.
Bier, M.C.J., Poletto, S., Soccol, V.T., Soccol, C.R., Medeiros, A.B.P., 2011. Isolation and
screening of microorganisms with potential for biotransformation of terpenic sub-
Conflict of interest strates. Braz. Arch. Biol. Technol. 54, 1019–1026.
Bier, M.C.J., Medeiros, A.B.P., Soccol, C.R., 2016. Biotransformation of limonene by
endophytic fungus using synthetic and orange residue-based media. Fungal Biology
Authors declare that they do not have conflict of interest. 121, 137–144.
Biji, H.L., Schaap, A., 2004. Isolation of microbial oil. United States Patent 2004/
Acknowledgements 067574 A1.
Bodour, A.A., Dress, K.P., Maier, R.M., 2003. Distribution of biosurfactant-producing
bacteria in undisturbed and contaminated arid southwestern soils. Appl. Environ.
Authors acknowledge the funding agencies Conselho Nacional de Microbiol. 69, 3280–3287.
Desenvolvimento Científico e Tecnológico (CNPq – Process number Boels, I.C., Ramos, A., Kleerebezem, M., de Vos, W.M., 2001. Functional analysis of the
Lactococcus lactis galU and galE genes and their impact on sugar nucleotide and
460897/2014-4; 140513/2016-7), Fundação de Amparo à Pesquisa do
exopolysaccharide biosynthesis. Appl. Environ. Microbiol. 67, 3033–3040.
Estado de São Paulo (FAPESP – Process number 2013/18390-0), Brar, K.K., Sarma, A.K., Aslam, M., Polikarpov, I., Chadha, B.S., 2017. Potential of olea-
Fundação de Amparo à Pesquisa do Estado de Minas Gerais (Fapemig – ginous yeast Trichosporon sp., for conversion of sugarcane bagasse hydrolysate into
Process number CAG – APQ-01056-17) and Coordenação de biodiesel. Bioresour. Technol. 242, 161–168.
Burdock, G.A., 2010. Fenaroli’s Handbook of Flavor Ingredients, Sixth edition. CRC Press,
Aperfeiçoamento de Pessoal de Nível Superior (Capes). Boca Raton, FL.
Cai, Q., Zhang, B., Chen, B., Zhu, Z., Lin, Z., Cao, T., 2014. Screening of biosurfactant
References producers from petroleum hydrocarbon contaminated sources in cold marine en-
vironments. Mar. Pollut. Bull. 86, 402–410.
Cameotra, S.S., Makkar, R.S., 2010. Biosurfactant-enhanced bioremediation of hydro-
Abdellah, M., Ahcène, H., Benali, Y., Saad, B., Abdelmalek, B., 2014. Evaluation of biofilm phobic pollutants. Pure Appl. Chem. 82, 97–116.
formation by exopolysaccharide-producer strains of thermophilic lactic acid bacteria Canilha, L., Silva, D.D.V., Carvalho, W., Mancilha, M., 2006. Aditivos alimentares pro-
isolated from Algerian camel milk. Emir. J. Food Agric. 27, 513–521. duzidos por via fermentativa parte 3: polissacarídeos e enzimas. Rev. Anal. 20,
Adebayo-Tayo, B.C., Onilude, A.A., 2008. Screening of lactic acid bacteria strains isolated 332–341.
from some Nigerian fermented foods for EPS production. World Appl. Sci. J. 4, Chakraborty, S., Ghosh, M., Chakraborti, S., Jana, S., Sen, K.K., Kokare, C., Zhang, L.,
741–747. 2015. Biosurfactant produced from Actinomycetes nocardiopsis A17: characterization
Almeida, D.G., Silva, R.C.F.S., Brasileiro, P.P.F., Luna, J.M., Rufino, R.D., Sarubbo, L.A., and its biological evaluation. Int. J. Biol. Macromol. 79, 405–412.
2017. Commercial formulation of biosurfactant from yeast and its evaluation to use Chaudhry, V., Nautiyal, C.S., 2011. A high throughput method and culture medium for
in the petroleum industry. Chem. Eng. Trans. 57, 661–666. rapid screening of phosphate accumulating microorganisms. Bioresour. Technol. 102,
Aranda-Selverio, G., Barretto Penna, A.L., Campos-Sás, L.F., Dos Santos Junior, O., 8057.
Dalberto Vasconcelos, A.F., De Lourdes Corradi da Silva, M., et al., 2010. Cirigliano, M.C., Carman, G.M., 1984. Purification and characterization of liposan, a
Propriedades reológicas e efeito da adição de sal na viscosidade de exopolissacarídeos bioemulsifier from Candida lipolytica. Appl. Environ. Microbiol. 50, 846–850.
produzidos por bactérias do gênero Rhizobium. Quim. Nova 33, 895–899. Cui, Y., Fraser, C., Gardner, G., Huang, C.J., Reith, M., Windust, A.J., 2012. Isolation and
Arous, F., Triantaphyllidou, I.E., Mechichi, T., Azabou, S., Nasri, M., Aggelis, G., 2015. optimization of the oleaginous yeast Sporobolomyces roseus for biosynthesis of 13C
Lipid accumulation in the new oleaginous yeast Debaryomyces etchellsii correlates isotopically labelled 18-carbon unsaturated fatty acids and trans 18: 1 and 18: 2
with ascosporogenesis. Biomass Bioenergy 80, 307–315. derivatives through synthesis. J. Ind. Microbiol. Biotechnol. 39, 153–161.
Ayadi, I., Kamoun, O., Trigui-Lahiani, H., Hdiji, A., Gargouri, A., Belghith, H., Guerfali, Czaczyk, K., Myszka, K., 2007. Biosynthesis of extracellular polymeric substances (EPS)
M., 2016. Single cell oil production from a newly isolated Candida viswanathii Y-E4 and its role in microbial biofilm formation. Pol. J. Environ. Stud. 16, 799.
and agro-industrial by-products valorization. J. Ind. Microbiol. Biotechnol. 43, Dai, J.-Y., Cheng, L., He, Q.-F., Xiu, Z.-L., 2015. High acetoin production by a newly
336
isolated marine Bacillus subtilis strain with low requirement of oxygen supply. Bozonnet, S., O’Donohue, M.J., 2016. Design of chromogenic probes for efficient
Process Biochem. 50, 1730–1734. screening and evaluation of feruloyl esterase-like activities. J. Mol. Catal. B Enzym.
Das, P., Mukherjee, S., Sen, R., 2008. Antimicrobial potential of a lipopeptide bio- 126, 24.
surfactant derived from a marine Bacillus circulans. J. Appl. Microbiol. 104, Grand View Research Inc., 2014. Biosurfactant Market Analysis By Product
1675–1684. (Rhamnolipids, Sophorolipids, MES, APG, Sorbitan Esters, Sucrose Esters) And
Das, P., Mukherjee, S., Sivapathasekaran, C., Sen, R., 2010. Microbial surfactants of Segment Forecast to 2020. Available at: http://www.grandviewresearch.com/
marine origin: potentials and prospects. In: Biosurfactants. Springer, pp. 88–101. industry-analysis/biosurfactants-industry.
De Carvalho, C.C., da Fonseca, M.M.R., 2006. Biotransformation of terpenes. Biotechnol. Gudiña, E.J., Rangarajan, V., Sen, R., Rodrigues, L.R., 2013. Potential therapeutic ap-
Adv. 24, 134. plications of biosurfactants. Trends Pharmacol. 34, 667–675.
Deeba, F., Pruthi, V., Negi, Y.S., 2017. Fostering triacylglycerol accumulation in novel Guichard, H., Bonnarme, P., 2005. Development and validation of a plate technique for
oleaginous yeast Cryptococcus psychrotolerans IITRFD utilizing groundnut shell for screening of microorganisms that produce volatile sulfur compounds. Anal. Biochem.
improved biodiesel production. Bioresour. Technol. 242, 113–120. 338, 299–305.
Deepika, K.V., Nagaraju, G.P., Bramhachari, P.V., 2016. Optimization of cultural condi- Habibi, H., Khosravi-Darani, K., 2017. Effective variables on production and structure of
tions for marine microbial biosurfactant production: future prospects from untapped xanthan gum and its food applications: a review. Biocatal. Agric. Biotechnol. 10,
marine resources. In: Marine Pollution and Microbial Remediation. Springer, pp. 130–140.
105–128. Hassanshahian, M., 2014. Isolation and characterization of biosurfactant producing
Demyttenaere, J.C.R., Vanoverschelde, J., de Kimpe, N., 2004. Biotransformation of (R)- bacteria from Persian Gulf (Bushehr provenance). Mar. Pollut. Bull. 86, 361–366.
(+)- and (S)-(-)- citronellol by Aspergillus sp. and Penicillium sp. and the use of solid- He, M., Hu, Q., Gou, X., Liu, X., Li, Q., Pan, K., Zhu, Q., Wu, J., 2010. Screening of
phase microextraction for screening. J. Chromatogr. A 1027, 137–146. oleaginous yeast with xylose assimilating capacity for lipid and bio-ethanol produc-
Deng, M.C., Li, J., Hong, Y.H., Xu, X.M., Chen, W.X., Yuan, J.P., Peng, J., Yi, M., Wang, tion. Afr. J. Biotechnol. 9, 8392–8397.
J.H., 2016. Characterization of a novel biosurfactant produced by marine hydro- Hong, S.-P., Sharpe, P.L., Xue, Z., Yadav, N.S., Zhang, H., Zhu, Q.Q., 2012. Recombinant
carbon-degrading bacterium Achromobacter sp. HZ01. J. Appl. Microbiol. 120, microbial host cells for high eicosapentaenoic acid production. United States Patent
889–899. 2012/052537 A1.
Desai, J.D., Banat, I.M., 1997. Microbial production of surfactants and their commercial Hua, D., Ma, C., Lin, S., Song, L., Deng, Z., Maomy, Z., 2007. Biotransformation of iso-
potential. Microbiol. Mol. Biol. Rev. 61, 47–64. eugenol to vanillin by a newly isolated Bacillus pumilus strain: identification of major
Dionísio, A.P., Molina, G., Carvalho, D.S., Santos, R., Bicas, J.L., Pastore, G.M., 2012. metabolites. J. Biotechnol. 130, 463–470.
Natural flavorings from biotechnology for food and beverages. In: Baines, D., Seal, R. Huang, C., Chen, X., Xiong, L., Chen, X., Ma, L., Chen, Y., 2013. Single cell oil production
(Eds.), Natural Food Additives, Ingredients and Flavourings. Woodhead Publishing from low-cost substrates: the possibility and potential of its industrialization.
Series in Food Science, Technology and Nutritionpp. 231–259. Biotechnol. Adv. 31, 129–139.
Dong, H., Xia, W., Dong, H., She, Y., Zhu, P., Liang, K., Zhang, Z., Liang, C., Song, Z., Sun, Huembelin, M., Beekwilder, M.J., Gerardus, J., Kierkels, T., 2014. Rhodobacter for
S., Zhang, G., 2016. Rhamnolipids produced by indigenous Acinetobacter junii from Preparing Terpenoids. (WO2014014339A2).
petroleum reservoir and its potential in enhanced oil recovery. Front. Microbiol. 7. Ikuo, K., Kenji, Y., & Keiko, H., 2017. U.S. Patent No. WO2017146213 (A1). Washington,
Donot, F., Fontana, A., Baccou, J.C., Schorr-galindo, S., 2012. Microbial exopoly- DC: U.S. Patent and Trademark Office.
saccharides: main examples of synthesis, excretion, genetics and extraction. Jazini, M.H., Fereydouni, E., Karimi, K., 2017. Microbial xanthan gum production from
Carbohydr. Polym. 87, 951–962. alkali-pretreated rice straw. RSC Adv. 7, 3507–3514.
Donot, F., Fontana, A., Baccou, J.C., Strub, C., Schorr-Galindo, S., 2014. Single cell oils Jemil, N., Ayed, H.B., Hmidet, N., Nasri, M., 2016. Characterization and properties of
(SCOs) from oleaginous yeasts and moulds: production and genetics. Biomass biosurfactants produced by a newly isolated Bacillus methylotrophicus DCS1 and their
Bioenergy 68, 135–150. applications in enhancing solubility of hydrocarbon. World J. Microbiol. Biotechnol.
Duarte, S.H., De Andrade, C.C.P., Ghiselli, G., Maugeri, F., 2013. Exploration of Brazilian 32, 175.
biodiversity and selection of a new oleaginous yeast strain cultivated in raw glycerol. Jemil, N., Manresa, A., Rabanal, F., Ayed, H.B., Hmidet, N., Nasri, M., 2017. Structural
Bioresour. Technol. 138, 377–381. characterization and identification of cyclic lipopeptides produced by Bacillus me-
Ebadipour, N., Lotfabad, T.B., Yaghmaei, S., RoostAazad, R., 2016. Optimization of low- thylotrophicus DCS1 strain. J. Chromatogr. B 1060, 374–386.
cost biosurfactant production from agricultural residues through response surface Jeong, D., Kim, D., Kang, I., Kim, H., Song, K., Kim, H., et al., 2017. Characterization and
methodology. Prep. Biochem. Biotechnol. 46, 30–38. antibacterial activity of a novel exopolysaccharide produced by Lactobacillus kefir-
Elazzazy, A.M., Abdelmonei, T.S., Almaghrabi, O.A., 2015. Isolation and characterization anofaciens DN1 isolated from ke fi r. Food Control 78, 436–442.
of biosurfactant production under extreme environmental conditions by alkali-halo- Ji, F., Li, L., Ma, S., Wang, J., Bao, Y., 2016. Production of rhamnolipids with a high
thermophilic bacteria from Saudi Arabia. Saud J. Biol. Sci. 22, 466–475. specificity by Pseudomonas aeruginosa M408 isolated from petroleum-contaminated
Emmerstorfer-Augustin, A., Moser, S., Pichler, H., 2016. Screening for improved iso- soil using olive oil as sole carbon source. Ann. Microbiol. 66, 1145–1156.
prenoid biosynthesis in microorganisms. J. Biotechnol. 235, 112–120. Jin, M., Slininger, P.J., Dien, B.S., Waghmode, S., Moser, B.R., Orjuela, A., Sousa, L. da C.,
Etschmann, M.M.W., Sell, D., Schrader, J., 2003. Screening of yeasts for the production of Balan, V., 2015. Microbial lipid-based lignocellulosic biorefinery: feasibility and
the aroma compound 2-phenylethanol in a molasses-based medium. Biotechnol. Lett. challenges. Trends Biotechnol. 33, 43–54.
25, 531–536. Joy, S., Rahman, P.K.S.M., Sharma, S., 2017. Biosurfactant production and concomitant
Fanaro, G.B., Fontanille, P., Maróstica Jr., M.R., Bicas, J.L., 2016. Biotechnological aroma hydrocarbon degradation potentials of bacteria isolated from extreme and hydro-
compounds. In: Bicas, J.L., Maróstica Jr.M.R., Pastore, G.M. (Eds.), Biotechnological carbon contaminated environments. Chem. Eng. J. 317, 232–241.
Production of Natural Ingredientes for Food Industry. Bentham Science Publishers, Jullesson, D., David, F., Pfleger, B., Nielsen, J., 2015. Impact of synthetic biology and
pp. 255–287. metabolic engineering on industrial production of fine chemicals. Biotechnol. Adv.
Faria, A.F., Teodoro-Martinez, D.S., Barbosa, G.N.O., Vaz, B.G., Silva, I.S., Garcia, J.S., 33, 1395.
Tótola, M.R., 2011. Production and structural characterization of surfactin (C14/ Kaci, Y., Heyraud, A., Barakat, M., Heulin, T., 2005. Isolation and identification of an EPS-
Leu7) produced by Bacillus subtilis isolate LSFM-05 grown on raw glycerol from the producing Rhizobium strain from arid soil (Algeria): characterization of its EPS and
biodiesel industry. Process Biochem. 46, 1951–1957. the effect of inoculation on wheat rhizosphere soil structure. Res. Microbiol. 156,
Felipe, L.O., de Oliveira, A.M., Bicas, J.L., 2017. Bioaromas – perspectives fos sustainable 522–531.
development. Trends Food Sci. Technol. 62, 141–153. Kennedy, J., O’Leary, N.D., Kiran, G.S., Morrissey, J.P., O’Gara, F., Selvin, J., Dobson,
Ferreira, A., Vecino, X., Ferreira, D., Cruz, J.M., Moldes, A.B., Rodrigues, L.R., 2017. A.D., 2011. functional metagenomic strategies for the discovery of novel enzymes
Novel cosmetic formulations containing a biosurfactant from Lactobacillus paracasei. and biosurfactants with biotechnological applications from marine ecosystems. J.
Colloids Surf. B: Biointerfaces 155, 522–529. Appl. Microbiol. 111, 787–789.
Finco, A.M. de O., Mamani, L.D.G., de Carvalho, J.C., de Melo Pereira, G.V., Thomaz- Keskin, N.O.S., Han, D., Ozkan, A.D., Angun, P., Umu, O.C.O., Tekinay, T., 2015.
Soccol, V., Soccol, C.R., 2016. Technological trends and market perspectives for Production and structural characterization of biosurfactant produced by newly iso-
production of microbial oils rich in omega-3. Crit. Rev. Biotechnol. 37, 656–671. lated Staphylococcus xylosus STF1 from petroleum contaminated soil. J. Pet. Sci. Eng.
Focus on Surfactants, 2014. Global Biosurfactants Market to Reach $2308.8 M by 2020. 133, 689–694.
7. pp. 2–3. Khopade, A., Biao, R., Liu, X., Mahadik, K., Zhang, L., Kokare, C., 2012. Production and
Franzetti, A., Gandolfi, I., Bestetti, G., Smyth, T.J.P., Banat, I.M., 2010. Production and stability studies of the biosurfactant isolated from marine Nocardiopsis sp. B4.
applications of trehalose lipid biosurfactants. Eur. J. Lipid Sci. Technol. 112, Desalination 285, 198–204.
617–627. Khot, M., Kamat, S., Zinjarde, S., Pant, A., Chopade, B., Ravikumar, A., 2012. Single cell
Freitas, F., Alves, V.D., Reis, M.A.M., 2011. Advances in bacterial exopolysaccharides: oil of oleaginous fungi from the tropical mangrove wetlands as a potential feedstock
from production to biotechnological applications. Trends Biotechnol. 29, 388–398. for biodiesel. Microb. Cell Factories 11, 1–13.
Furukawa, H., Morita, H., Yoshida, T., Nagasawa, T., 2003. Conversion of isoeugenol into Kim, Y. S., Yoon, S. K., & Shin, E. J., 2017. U.S. Patent No. KR101682101. Washington,
vanillic acid by Pseudomonas putida I58 cells exhibiting high isoeugenol-degrading DC: U.S. Patent and Trademark Office.
activity. J. Biosci. Bioeng. 96, 401–403. Kiran, G.S., Thomas, T.A., Selvin, J., Sabarathnam, B., Lipton, A.P., 2010. Optimization
Gandhimathi, R., Kiran, G.S., Hema, T.A., Selvin, J., Raviji, T.R., Shanmughapriya, S., and characterization of a new lipopeptide biosurfactant produced by marine
2009. Production and characterization of lipopeptide biosurfactant by a sponge-as- Brevibacterium aureum MSA13 in solid state culture. Bioresour. Technol. 101,
sociated marine actinomycetes Nocardiopsis alba MSA10. Bioprocess Biosyst. Eng. 32, 2389–2396.
825–835. Kiran, G.S., Priyadharsini, S., Sajayan, A., Priyadharsini, G.B., Poulose, N., Selvin, J.,
Gesheva, V., Stackebrandt, E., Vasileva-Tonkova, E., 2010. Biosurfactant production by 2017. Production of lipopeptide biosurfactant by a marine Nesterenkonia sp. and its
halotolerant Rhodococcus fascians from Casey station, Wilkes Land, Antarctica. Curr. application in food industry. Front. Microbiol. 8 (article 1138), 1–11.
Microbiol. 61, 112–117. Kitcha, S., Cheirsilp, B., 2011. Screening of oleaginous yeasts and optimization for lipid
Gherbovet, O., Fauré, R., Ferreira, F., Durand, J., Ragon, M., Hostyn, G., Record, E., production using crude glycerol as a carbon source. Energy Procedia 9, 274–282.
337
Kitcha, S., Cheirsilp, B., 2013. Enhancing lipid production from crude glycerol by newly Morita, T., Fukuoka, T., Imura, T., Hirose, N., Kitamoto, D., 2012. Isolation and screening
isolated oleaginous yeasts: strain selection, process optimization, and fed-batch of glycolipid biosurfactant producers from sugarcane. Biosci. Biotechnol. Biochem.
strategy. Bioenergy Resour. 6, 300–310. 76, 1788–1791.
Kitcha, S., Cheirsilp, B., 2014. Bioconversion of lignocellulosic palm byproducts into Moscovici, M., 2015. Present and future medical applications of microbial exopoly-
enzymes and lipid by newly isolated oleaginous fungi. Biochem. Eng. J. 88, 95–100. saccharides. Front. Microbiol. 6, 1–11. https://doi.org/10.3389/fmicb.2015.01012.
Koskinen, P., Tanner, R., 2012. Integrated process system. United States Patent 2012/ Mouafi, F.E., Elsoud, M.M.A., Moharam, M.E., 2016. Optimization of biosurfactant pro-
159840 A1. duction by Bacillus brevis using response surface methodology. Biotechnol. Rep. 9,
Kraisintu, P., Yongmanitchai, W., Limtong, S., 2010. Selection and optimization for lipid 31–37.
production of a newly isolated oleaginous yeast Rhodosporidium toruloides DMKU3- Mukherjee, S., Das, P., Sen, R., 2006. Towards commercial production of microbial sur-
TK16. Kasetsart J. 44, 436–445. factants. Trends Biotechnol. 24, 509–515.
Kumar, A.S., Mody, K., Jha, B., 2007. Bacterial exopolysaccharides - A perception. J. Basic Nalini, S., Parthasarathi, R., 2014. Production and characterization of rhamnolipids
Microbiol. 47 (2), 103–117. https://doi.org/10.1002/jobm.200610203. produced by Serratia rubidaea SNAU02 under solid-state fermentation and its appli-
Lahaye, É., 2006. Rôle structurant des exopolysaccharides dans un biofilm bactérien. cation as biocontrol agent. Bioresour. Technol. 173, 231–238.
Available at: http://www.theses.fr/2006LORIS071. Nwodo, U.U., Green, E., Okoh, A.I., 2012. Bacterial exopolysaccharides: functionality and
Lawrance, A., Balakrishnan, M., Joseph, T.C., Sukumaran, D.P., Valsalan, V.N., Gopal, D., prospects. Int. J. Mol. Sci. 13, 14002–14015.
Ramalingam, K., 2014. Functional and molecular characterization of a lipopeptide Ochsenreither, K., Glück, C., Stressler, T., Fischer, L., Syldatk, C., 2016. Production
surfactant from the marine sponge-associated eubacteria Bacillus licheniformis NIOT- strategies and applications of microbial single cell oils. Front. Microbiol. 7.
AMKV06 of Andaman and Nicobar Islands, India. Mar. Pollut. Bull. 82, 76–85. Ohmart, C.P., Thomas, J.R., Bubela, B., 1988. Surfactant-producing microorganisms
Leffingwell & Associates Flavor & Fragrance Industry Leaders. http://www.leffingwell. isolated from the gut of a Eucalyptus-Feeding Sawfly, Perga affinis affinis. Oecologia
com/top_10.htm Accessed 03.07.2017. 77, 140–142.
Leffingwell, J.C., Leffingwell, D., 2015. Biotechnology – conquests and challenges in Onwosi, C.O., Odibo, F.J., 2012. Effects of carbon and nitrogen sources on rhamnolipid
flavors & fragrances. Leffingwell Reports 7, 1–11. biosurfactant production by Pseudomonas nitroreducens isolated from soil. World J.
Li, S.L., Feng, S.L., Li, Z.T., Xu, H., Yu, Y.P., Qiao, D.R., Cao, Y., 2011. Isolation, identi- Microbiol. Biotechnol. 28, 937–942.
fication and characterization of oleaginous fungi from the soil of Qinghai Plateau that Ortega-Morales, B.O., Santiago-García, J.L., Chan-Bacab, M.J., Moppert, X., Miranda-
utilize D-xylose. Afr. J. Biotechnol. 5, 2075–2081. Tello, E., Fardeau, M.L., Carrero, J.C., Bartolo-Pérez, P., Valadéz-Gonzalez, A.,
Li, J., Deng, M., Wang, Y., Chen, W., 2016. Production and characteristics of biosurfactant Guezennec, J., 2007. Characterization of extracellular polymers synthesized by tro-
produced by Bacillus pseudomycoides BS6 utilizing soybean oil waste. Int. Biodeterior. pical intertidal biofilm bacteria. J. Appl. Microbiol. 102, 254–264.
Biodegrad. 112, 72–79. Pacwa-Płociniczak, M., Płaza, G.A., Piotrowska-Seget, Z., Cameotra, S.S., 2011.
Liaud, N., Navarro, D., Vidal, N., Sigoillot, J.C., Raouche, S., 2014. High throughput Environmental applications of biosurfactants: recent advances. Int. J. Mol. Sci. 12,
automated colorimetric method for the screening of L-lactic acid producing micro- 633–654.
organisms. MethodsX 1, 254. Palmerín-Carreño, D.M., Rutiaga-Quiñones, O.M., Calvo, J.R.V., Prado-Barragán, A.,
Liu, G.-Q., Lin, Q.-L., Jin, X.-C., Wang, X., Zhao, Y., 2010. Screening and fermentation Huerta-Ochoa, S., 2015. Screening of microorganisms for bioconversion of (+)-va-
optimization of microbial lipid-producing molds from forest soils. Afr. J. Biotechnol. lencene to (+)- nootkatone. LWT Food Sci. Technol. 64, 768–793.
4, 1462–1468. Pan, L.-X., Yang, D.-F., Shao, L., Li, W., Chen, G.-G., Liang, Z.-Q., 2009. Isolation of the
Longo, M.A., Sanromán, M.A., 2006. Production of food aroma compounds. Food oleaginous yeasts from the soil and studies of their lipid-producing capacities. Food
Technol. Biotechnol. 44, 335–353. Technol. Biotechnol. 47, 215–220.
Ma, X., Li, H., Song, X., 2012. Surface and biological activity of sophorolipid molecules Paulino, B.N., Pessôa, M.G., Mano, M.C.R., Molina, G., Neri-Numa, I.A., Pastore, G.M.,
produced by Wickerhamiella domercqiae var. sophorolipid CGMCC1576. J. Colloid 2016. Current status in biotechnological production and applications of glycolipids
Interface Sci. 376, 165–172. biosurfactants. Appl. Microbiol. Biotechnol. 100, 10265–10293.
Madhuri, K.V., Prabhakar, K.V., 2014. Microbial exopolysaccharides: biosynthesis and Pawar, S.T., Bhosale, A.A., Gawade, T.B., Nale, T.R., 2013. Isolation, Screening and
potential applications. Orient. J. Chem. 30 (3). Optimization of Exopolysaccharide Producing Bacterium From Saline Soil. Available
Mamatha, S.S., Halami, P.M., Venkateswaran, G., 2010. Identification and characteriza- online at. www.scholarsresearchlibrary.com vol. 3. pp. 24–31.
tion of the n-6 fatty acid-producing Mucor rouxii native isolate CFR-G15. Eur. J. Lipid Pessôa, M.G., Paulino, B.N., Mano, M.C.R., Neri-Numa, I.A., Molina, G., Pastore, G.M.,
Sci. Technol. 112, 380–389. 2017. Fusarium species – a promising tool box for industrial biotechnology. Appl.
Maróstica Jr., M.R., Pastore, G.M., 2007. Production of R-(+)-α-terpineol by the bio- Microbiol. Biotechnol. 101 (9), 3493–3511.
transformation of limonene from orange essential oil, using cassava waste water as Pimentel, M.R., Molina, G., Dionísio, A.P., Maróstica Jr., M.R., Pastore, G.M., 2011. The
medium. Food Chem. 101, 345–350. use of endophytes to obtain bioactive compounds and their application in bio-
Mnif, I., Ghribi, D., 2015. Review lipopeptides biosurfactants: mean classes and new in- transformation process. Biotechnol. Res. Int. 2011, 11 pages.
sights for industrial, biomedical, and environmental applications. Biopolymers 104, Pizon, N., Ju, L.K., 2009. Improved detection of rhamnolipid production using agar plates
129–147. containing Methylene blue and Cetyltrimethyl ammonium bromide. Biotechnol. Lett.
Mnif, I., Besbes, S., Ellouze-Ghorbel, R., Ellouze-Chaabouni, S., Ghribi, D., 2013. 31, 1583–1588.
Improvement of bread dough quality by Bacillus subtilis SPB1 biosurfactant addition: Pogacic, T., Maillard, M.B., Leclerc, A., Hervé, C., Chuat, V., Yee, A., Valence, F., Thierry,
optimized extraction using response surface methodology. J. Sci. Food Agric. 93, A., 2015. A methodological approach to screen diverse cheese-related bacteria for
3055–3064. their ability to produce aroma compounds. Food Microbiol. 46, 145–153.
Mohanram, R., Jaqtap, C., Kumar, P., 2016. Isolation, screening, and characterization of Polburee, P., Yongmanitchai, W., Lertwattanasakul, N., Ohashi, T., Fujiyama, K., Limtong,
surface-active agent-producing, oil-degrading marine bacteria of Mumbai Harbor. S., 2015. Characterization of oleaginous yeasts accumulating high levels of lipid
Mar. Pollut. Bull. 105, 131–138. when cultivated in glycerol and their potential for lipid production from biodiesel-
Molbase, 2015. Chemical Search and Share. http://www.molbase.com, Accessed date: 7 derived crude glycerol. Fungal Biol. 19, 1194–1204.
March 2017. Randhawa, K.K.S., Rahman, P.K.S.M., 2014. Rhamnolipid biosurfactants- past, presente,
Molina, G., Bicas, J.L., Maróstica Jr., M.R., Pastore, G.M., 2012. Compostos de Aroma. In: and future scenario of global market. Front. Microbiol. 5, 454.
Pastore, G.M., Bicas, J.L., Maróstica Jr.M.R. (Eds.), Biotecnologia de Alimentos. Ratledge, C., 2013. Microbial oils: an introductory overview of current status and future
Atheneu, pp. 273–296. prospects. Ocl 20, D602. https://doi.org/10.1051/ocl/2013029.
Molina, G., Pinheiro, M., Pimentel, M.R., Santos, R., Pastore, G.M., 2013. Monoterpene Rawal, P.M., Chauhan, P.B., Prajapati, H., Gahlout, M., 2016. Evaluation of cultivation
bioconversion for the production of aroma compounds by fungi isolated from condition for enhanced production of exopolysaccharide by bacterial isolate P 11
Brazilian fruits. Food Sci. Biotechnol. 22, 999–1006. under submerged culture condition. Int. J. Adv. Res. Biol. Sci. 3, 183–190.
Molina, G., Pessôa, M.G., Pimentel, M.R., Pelissari, F.M., Bicas, J.L., Pastore, G.M., 2014. Ruas-Madiedo, P., de los Reyes-Gavilán, C.G., 2005. Invited review: methods for the
Production of natural flavor compounds using monoterpenes as substrates. In: screening, isolation, and characterization of exopolysaccharides produced by lactic
Jinnah, H. (Ed.), New Developments in Terpene Research. Nova Publishers, New acid bacteria. J. Dairy Sci. 88, 843–856. https://doi.org/10.3168/jds.S0022-
York, pp. 1–24. 0302(05)72750-8.
Molina, G., Abrahão, M.R.E., Pessôa, M.G., Bution, M.L., Paulino, B.N., Néri-Numa, I.A., Rikalovic, M.G., Abdel-Mawgoud, A.M., Déziel, E., Gojgic-Cvijovic, G.Dj., Nestorovic, Z.,
Pastore, G.M., 2016. Industrial additives obtained through microbial biotechnology: Vrvic, M.M., Karadzic, I.M., 2013. Comparative analysis of rhamnolipids from novel
bioflavors and Biocolorants. In: Gupta, V.K., Sharma, G.D., Tuohy, M.G., Gaur, R. environmental isolates of Pseudomonas aeruginosa. J. Surfactant Deterg. 16, 673–682.
(Eds.), The Handbook of Microbial Bioresources. CABI Publishing, Boston, pp. Roca, C., Lehmann, M., Torres, C.A., Baptista, S., Gaudêncio, S.P., Freitas, F., Reis, M.A.,
549–566. 2016. Exopolysaccharide production by a marine Pseudoalteromonas sp. strain iso-
Morita, T., Takashima, M., Fukuoka, T., Konishi, M., Imura, T., Kitamoto, D., 2010. lated from Madeira Archipelago ocean sediments. New Biotechnol. 33, 460–466.
Isolation of basidiomycetous yeast Pseudozyma tsukubaensis and production of gly- Rodrígues-Bustamante, E., Sánches, S., 2007. Microbial production of C13-norisoprenoids
colipid biosurfactant, a diasteomer type of mannosylerythritol lipid-B. Appl. and other aroma compounds via carotenoid cleavage. Crit. Rev. Microbiol. 33,
Microbiol. Biotechnol. 88, 679–688. 211–230.
Morita, T., Ogura, Y., Takashima, M., Hirose, N., Fukuoka, T., Imura, T., Kondo, Y., Ron, E.Z., Rosenberg, E., 2001. Natural roles of biosurfactants. Environ. Microbiol. 3,
Kitamoto, D., 2011a. Isolation of Pseudozyma churashimaensis sp. Nov., a novel usti- 229–236.
laginomycetous yeast species as a producer of glycolipid biosurfactants, mannosy- Rosenberg, E., Rubinovitz, C., Legmann, R., Ron, E.Z., 2001. Purification and chemical
lerythritol lipids. J. Biosci. Bioeng. 112, 137–144. properties of Acinetobacter calcoaceticus As Biodispersan. Appl. Environ. Microbiol.
Morita, T., Ishibashi, Y., Hirose, N., Wada, K., Takahashi, M., Fukuoka, T., Imura, T., 54, 323–326.
Sakai, H., Abe, M., Kitamoto, D., 2011b. Production and characterization of a gly- Rottava, I., Toniazzo, G., Cortina, P.F., Martello, E., Grando, C.E., Lerin, L.A., 2010.
colipid biosurfactant, mannosylerythritol lipid B, from sugarcane juice by Ustilago Screening of microorganisms for bioconversion of (-)-β-pinene and R-(+)-limonene
scitaminea NBRC 32730. Biosci. Biotechol. Biochem. 75, 1371–1376. to α-terpineol. LWT Food Sci. Technol. 43, 1128–1131.
338
Rottava, I., Cortina, P.F., Martelo, E., Cansian, R.L., Toniazzo, G., Antunes, O.A.C., 2011. 1909–1921.
Optimization of α-terpineol production by biotransformation of R-(+)- limonene and Toren, A., Navon-Venezia, S., Ron, E.Z., 2001. Emulsifying activity of purified alasan
(-)-β-pinene. Appl. Biochem. Biotechnol. 164, 514–523. proteins from Acinetobacter radioresistens. Appl. Environ. Microbiol. 67, 1102–1106.
Rühmann, B., Schmid, J., Sieber, V., 2015. Methods to identify the unexplored diversity of Trimbur, D.E., Im, C.-S., Dillon, H.F., Day, A.G., Franklin, S., Coragliotti, A., 2008.
microbial exopolysaccharides. Front. Microbiol. 6, 565. Production of Oil in Microorganisms. World Intellectual Property Organization 2008/
Sabaté, D.C., Carrillo, L., Audisio, M.C., 2009. Inhibition of Paenibacillus larvae and 151149 A2.
Ascosphaera apis by Bacillus subtilis isolated from honeybee gut and honey samples. Trimbur, D.E., Im, C.-S., Dillon, H.F., Day, A.G., Franklin, S., Coragliotti, A., 2016.
Res. Microbiol. 160, 193–199. Renewable diesel and jet fuel from microbial sources. United States Patent 9,434,
Sankh, S., Thiru, M., Saran, S., Rangaswamy, V., 2013. Biodiesel production from a newly 909 B2.
isolated Pichia kudriavzevii strain. Fuel 106, 690–696. Trytek, M., Jedrzejewski, K., Fiedurek, J., 2015. Bioconversion of alpha-pineneby a novel
Saran, S., Mathur, A., Dalal, J., Saxena, R.K., 2017. Process optimization for cultivation cold-adapted fungus Chrysosporium pannorum. J. Ind. Microbiol. Biotechnol. 42,
and oil accumulation in an oleaginous yeast Rhodosporidium toruloides A29. Fuel 188, 181–188.
324–331. UBIC Consulting, 2014. The World Biotechn Flavours Market. http://ubicconsulting.com/
Sarma, S.J., Dhillon, G.S., Hegde, K., Brar, S.K., Verma, M., 2014. Utilization of agro- wp-content/uploads/2015/08/Biotech-Flavor.pdf Accesses 03.07.2017.
industrial waste for the production of aroma compounds and fragrances. In: Brar, S. Uzoigwe, C., Burgess, J.G., Ennis, C.J., Rahman, P.K.S.M., 2015. Bioemulsifiers are not
(Ed.), Biotransformation of Waste Biomass into High Value Biochemicals. Springer biosurfactants and require different screening approaches. Front. Microbiol. 6
Science+Business Media, New York, pp. 99–115. (245), 1–6.
Satpute, S.K., Banat, I.M., Dhakephalkar, P.K., Banpurkar, A.G., Chopade, B.A., 2010. Vainio, H., Sipponen, M., Laakso, T., Pastinen, O., Lehtomäki, I., Kahelin, H., Koskinen,
Biosurfactants, bioemulsifiers and exopolysaccharides from marine microorganisms. P., Laamanen, M., 2016. Method for producing single cell oil from lignocellulosic
Biotechnol. Adv. 28, 436–450. materials. United States Patent 2016/304914 A1.
Schempp, F.M., Drummond, L., Buchhaupt, M., Schrader, J., 2017. Microbial cell factories Van Elsas, J.D., Boersma, F.G.H., 2011. A review of molecular methods to study the
for the production of terpenoid flavor and fragrance compounds. J. Agric. Food microbiota of soil and the mycosphere. Eur. J. Soil Biol. 47, 77.
Chem. 14, 2247–2258 In press. Van Geel-Schutten, G.H., Flesch, F., Ten Brink, B., Smith, M.R., Dijkhuizen, L., 1998.
Selvin, J., Shanmughapriya, S., Gandhimathi, R., Kiran, G.S., Ravji, T.R., 2009. Screening and characterization of Lactobacillus strains producing large amounts of
Optimization and production of novel antimicrobial agents from sponge associated exopolysaccharides. Appl. Microbiol. Biotechnol. 50, 697–703.
marine actinomycetes Nocardiopsis dassonvillei MAD08. Appl. Microbiol. Biotechnol. Vespermann, K.A., Paulino, B.N., Barcelos, M.C., Pessôa, M.G., Pastore, G.M., Molina, G.,
83, 435–445. 2017. Biotransformation of α-and β-pinene into flavor compounds. Appl. Microbiol.
Shimoni, E., Ravid, U., Shoham, Y., 2000. Isolation of a Bacillus sp. capable of trans- Biotechnol. 101, 1805.
forming isoeugenol to vanillin. J. Biotechnol. 78, 1–9. Vieira, J.P.F., Ienczak, J.L., Rossell, C.E.V., Pradella, J.G.C., Franco, T.T., 2014. Microbial
Sidorova, N.A., Voronov, P.D., 2016. Results of the study of mutagenic effects of micro- lipid production: screening with yeasts grown on Brazilian molasses. Biotechnol. Lett.
bial polysaccharides. Eur. J. Mol. Biotechnol. 12, 85–90. https://doi.org/10.13187/ 36, 2433–2442.
ejmb.2014.4.72. Vijayendra, S.V.N., Palanivel, G., Mahadevamma, S., Tharanathan, R.N., 2008. Physico-
Singh, R.S., Saini, G.K., Kennedy, J.F., 2008. Pullulan: microbial sources, production and chemical characterization of an exopolysaccharide produced by a non-ropy strain of
applications. Carbohydr. Polym. 73, 515–531. https://doi.org/10.1016/j.carbpol. Leuconostoc sp. CFR 2181 isolated from dahi, an Indian traditional lactic fermented
2008.01.003. milk product. Carbohydr. Polym. 72, 300–307.
Sitepu, I.R., Garay, L.A., Sestric, R., Levin, D., Block, D.E., German, J.B., Boundy-Mills, Vinarta, S.C., Angelicola, M.V., Barros, J.M., Fern, P.M., Aybar, M.J., de Figueroa, L.I.C.,
K.L., 2014. Oleaginous yeasts for biodiesel: current and future trends in biology and 2016. Oleaginous yeasts from Antarctica: screening and preliminary approach on
production. Biotechnol. Adv. 32, 1336–1360. lipid accumulation. J. Basic Microbiol. 56, 1–9.
Soberón-Chávez, G., Maier, R.M., 2011. Biosurfactants: a general overview. In: Soberón- Vyas, S., Chhabra, M., 2017. Isolation, identification and characterization of
Chávez, G. (Ed.), Biosurfactants, Microbiology Monographs 20. Springer-Verlag, Cystobasidium oligophagum JRC1: a cellulase and lipase producing oleaginous yeast.
Berlin, Germany, pp. 1–11. Bioresour. Technol. 223, 250–258.
Soccol, C.R., Dalmas Neto, C.J., Soccol, V.T., Sydney, E.B., da Costa, E.S.F., Medeiros, Walter, V., Syldatk, C., Hausmann, R., 2010. Screening concepts for the isolation of
A.B.P., Vandenberghe, L.P. de S., 2017. Pilot scale biodiesel production from mi- biosurfactant producing microorganisms. Biosurfactants 1–13.
crobial oil of Rhodosporidium toruloides DEBB 5533 using sugarcane juice: perfor- Wang, Y.S., Hu, Q.W., Zheng, X.C., Zhang, J.F., Zheng, Y.G., 2017. A spectrophotometric
mance in diesel engine and preliminary economic study. Bioresour. Technol. 223, screening method for avermectin oxidizing microorganisms. J. Microbiol. Methods
259–268. 135, 93.
Steensels, J., Meersman, E., Snoek, T., Saels, V., Verstrepen, K.J., 2014. Large-scale se- Xia, W., Du, Z., Cui, Q., Dong, H., Wang, F., He, P., Tang, Y., 2014. Biosurfactant pro-
lection and breeding to generate industrial yeasts with superior aroma production. duced by novel Pseudomonas sp. WJ6 with biodegradation of n-alkanes and polycyclic
Appl. Environ. Microbiol. 80, 6965–6975. aromatic hydrocarbons. J. Hazard. Mater. 276, 489–498.
Stephanopoulos, G., Tai, M., Chakraborty, S., 2013. Engineered microbes and methods for Xu, P., Hua, D., Lin, S., Song, L., Ma, C., Zhang, Z., 2009. Streptomyces strain and the
microbial oil production. United States Patent 2013/143282 A1. method of converting ferulic acid to vanillin by using the same. US Patent 2009/
Strobel, G., Daisy, B., 2003. Bioprospecting for microbial endophytes and their natural 0186399 A1.
products. Micobiol. Mol. Biol. Rev. 67, 491–502. Yadav, A.K., Manna, S., Pandiyan, K., Singh, A., Kumar, M., Chakdar, H., Kashyap, P.L.,
Sutherland, I.W., 1998. Novel and established applications of microbial polysaccharides. Srivastava, A.K., 2016. Isolation and characterization of biosurfactant producing
Tibtech 16, 41–46. Bacillus sp. from diesel fuel contaminated site. Microbiology 85, 56–62.
Subhash, G.V., Mohan, S.V., 2011. Biodiesel production from isolated oleaginous fungi Yamaguchi, H., Kosuge, Y., Ikeda, R., Doisaki, N., Sato, S., 2016. Microbial oil, producing
Aspergillus sp. using corncob waste liquor as a substrate. Bioresour. Technol. 102, method for microbial oil, concentrated microbial oil, and production method for
9286–9290. concentrated microbial oil. United States Patent 2016/317592 A1.
Subramanian, S., Yan, S., Tyagi, R.D., Surampalli, R.Y., 2010. Extracellular polymeric Yee, A.L., Maillard, M.B., Roland, N., Chuat, V., Leclerc, A., Pogacic, T., Valence, F.,
substances (EPS) producing bacterial strains of municipal wastewater sludge: isola- Thierry, A., 2014. Great interspecies and intraspecies diversity if dairy propioni-
tion, molecular identification, EPS characterization and performance for sludge set- bacteria in the production of cheese aroma compounds. Int. J. Food Microbiol. 191,
tling and dewatering. Water Res. 44, 2253–2266. 60–68.
Sutherland, I.W., 2001. Microbial polysaccharides from Gram-negative bacteria. Int. Yu, X., Zeng, J., Zheng, Y., Bule, M., Chen, S., 2014. Simultaneous saccharification and
Dairy J. 11 (9), 663–674. fermentation (SSF) of lignocellulosic biomass for single cell oil production by olea-
Tampitak, S., Louhasakul, Y., Cheirsilp, B., Prasertsan, P., 2015. Lipid production from ginous microorganisms. United States Patent 2014/234919 A1.
hemicellulose and holocellulose hydrolysate of palm empty fruit bunches by newly Zhao, L.Q., Sun, Z.H., Zheng, P., He, J.Y., 2006. Biotransformation of isoeugenol to va-
isolated oleaginous yeasts. Appl. Biochem. Biotechnol. 176, 1801–1814. nillin by Bacillus fusiformis CGMCC1347 with addition of resin HD-8. Process
Tanimura, A., Takashima, M., Sugita, T., Endoh, R., Kikukawa, M., Yamaguchi, S., Biochem. 41, 1673–1676.
Sakuradani, E., Ogawa, J., Shima, J., 2014. Selection of oleaginous yeasts with high Zhao, F., Zhou, J., Han, S., Ma, F., Zhang, Y., Zhang, J., 2016. Medium factors on
lipid productivity for practical biodiesel production. Bioresour. Technol. 153, anaerobic production of rhamnolipids by Pseudomonas aeruginosa SG and simplifying
230–235. medium for in situ microbial enhanced oil recovery applications. World J. Microbiol.
Tavares, L.F., Silva, P.M., Junqueira, M., Mariano, D.C., Nogueira, F.C., Domont, G.B., Biotechnol. 32, 54.
Freire, D.M., Neves, B.C., 2013. Characterization of rhamnolipids produced by wild- Zosim, Z., Gutnick, D.L., Rosenberg, E., 1982. Properties of hydrocarbon-in-water emul-
type and engineered Burkholderia kururiensis. Appl. Microbiol. Biotechnol. 97, sions stabilized by Acinetobacter RAG-1 emulsan. Biotechnol. Bioeng. 24, 281–292.
339

Biotechnology Advances

Uploaded by

Copyright:

Available Formats

Biotechnology Advances

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Biotechnology Advances

Uploaded by

Copyright:

Available Formats

Biotechnology Advances 37 (2019) 319–339

Contents lists available at ScienceDirect

Research review paper

Newly isolated microorganisms with potential application in biotechnology T

1. Introduction applications with the development of a spectrophotometric screening

Fig. 2. Chemical structure of aroma compounds obtained in fermentation processes.

n.d: not described.

Fig. 5. Fatty acids found in microbial oils produced by fermentation.

You might also like