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PRODUCTION OF AMYLASE FROM MICROBIAL SOURCES: A REVIEW

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DOI: 10.31032/IJBPAS/2021/10.4.1038

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 IJBPAS, April, Special Issue, 2021, 10(4): 341-350
ISSN: 2277–4998

PRODUCTION OF AMYLASE FROM MICROBIAL SOURCES: A

REVIEW

GOJIYA R1, ANDHARE P2, MARCHAWALA F2, BHATTACHARYA I2 AND

UPADHYAY D2*
1: Student, M. Sc. Microbiology, Parul Institute of Applied Sciences, Parul University, Post
Limda, Waghodia, Gujarat, 391760
2: Assistant Professor, Parul Institute of Applied Sciences, Parul University, Post Limda,
Waghodia, Gujarat, 391760
*Corresponding Author: Dr. Dhwani Upadhyay; E Mail:dhwani.updhyay82123@paruluniversity.ac.in;
Tel: +919558021474
nd
Received 22 Jan. 2021; Revised 24 Feb. 2021; Accepted 25th March 2021; Available online 1st April 2021
th

https://doi.org/10.31032/IJBPAS/2021/10.4.1038
ABSTRACT
Amylase is one of oldest enzyme. Amylase is naturally produced in humans by the salivary
glands and pancreas. It can also be found naturally in plants and animals. Amylase is an
enzyme that converts the hydrolysis of starch into sugar. Amylase divided into three types: 1)
α-amylase 2) ß-amylase and 3) γ-amylase. α-amylase: Alpha-amylase, is an enzyme EC
3.2.1.1 that hydrolyses alpha bonds of large,it is the major form of amylase found in humans
and other mammals. ß-amylasethe enzymes in the second group, exoamylases, are
exclusively cleaved a, 1-4. glycoside bonds such as beta-amylase or cleavage glycoside bonds
a,1-4 and a,1-6. Amyl glucosidase or glucoamylase and α-glucosidase. γ-amylase: Y1-
amylase cuts glycosidic α (1-6) compounds, in addition to purifying the final α (1-
4) glycosidic links in the non-limiting structure of amylose and amylopectin, express
glucose. Unlike other types of amylase, γ-amylase works best in acidic environments and
contains 3 pH high. Two major groups of microorganisms play an important role in
amylase.production, namely: bacteria and fungi. Starch is a substrate used in the production
of amylase. In betweentwo fermentation processes used in the production of amylase (e.g.,
immersed boiling and solid state fermentation), this has great benefits as it saves costs,

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Upadhyay D et al Review Article

produces less pollution, and is higher a lot of product. In this review, various applications,
production and factors affecting amylase activities are discussed.
Keywords: Enzyme, amylase, production, fermentation, SSF, SmF
INTRODUCTION
Amylase is perhaps the most seasoned beta-amylase or cleavage glycoside bonds
catalyst known, the historical backdrop of a,1-4 and a,1-6. [29] Amyl glucosidase or
amylase started in 1815 when Kirchhoff glucoamylase and α-glucosidase. Γ-
established the framework for the amylase: Y1-amylase cuts glycosidic α (1-
revelation of amylase, Kirchhoff performed 6) compounds, in addition to purifying the
an investigation, which changed over four final α (1-4) glycosidic links in the non-
pieces of water, two pieces of starch, and limiting structure of amylose and
malt into a starch glue. This glue started to amylopectin, express glucose. Unlike other
condense into sweet syrup; his outcomes types of amylase, γ-amylase works best in
demonstrated that gluten could change over acidic environments and contains 3 pH high
a bigger amount of starch into sugar. In [25]. Two major groups of microorganisms
1831, Erhard Friedrich Leuchs portrayed play an important role in amylase Synthetic
the hydrolysis of starch by spit, due to the catalysis however generally utilized was
presence of a protein in salivation, very awkward. The impediments that this
"ptyalin", an amylase [1]. In 1833, Anselme technique presents incorporate a
Payen and Jean-François Persoz further requirement for high temperature and
portray and confine diastase (amylase) in weight for catalysis and moderate
powder structure from grain malt, explicitness [28].
demonstrating it to be heat-labile [2] These restrictions were overwhelmed by
Compounds are organic impetuses which the utilization of compounds. Catalysts
are a vital segment of organic responses. work at milder conditions when contrasted
The utilization of substance impetuses has with that needed by synthetic impetuses for
been followed for an extremely long time. activity. Additionally, catalysts are
α-amylase: Alpha-amylase, is an enzyme exceptionally explicit and catalyze responses

EC 3.2.1.1 that hydrolyses alpha bonds of quicker than compound impetuses [1].

large,it is the major form of amylase found Compounds are currently being utilized in

in humans and other mammals. ß-amylase different areas of industry. They are

the enzymes in the second utilized in cleansers, paper industry,

group, exoamylases, are exclusively material industry, food industry, and

cleaved a, 1-4. glycoside bonds such as numerous other modern applications.

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Catalysts have been being used since old hydrolysis is carried out at higher
occasions [2] and they have been utilized in temperatures, the polymerization of D-
saccharification of starch, creation of glucose to iso-maltose is minimized [9].
refreshments like a lager, treatment of Amylase has been produced by main two
stomach related problems, and creation of methods;
cheddar from milk [4]. Among the  Submerged fermentation.
numerous catalysts that are generally  Solid state fermentation.
utilized α-Amylase has been in expanding Submerged fermentation:
request because of its pivotal part of starch Submerged fermentation is a method of
hydrolysis and the utilizations of this producing biomolecules that involves
hydrolytic activity. The accompanying submerging enzymes and other reactive
segments expound on the sorts of amylases compounds in a liquid such as alcohol, oil,
and their jobs in enzymatic responses. or nutrient broth. Submerged Fermentation
PRODUCTION OF AMYLASE: (SmF)/Liquid Fermentation (LF) uses free-
1) The growth of microorganisms is rapid flowing liquid substrates like molasses and
and this will in turn speed up the broths for fermentation. The procedure is
production of enzyme. Microorganisms are used for a number of reasons, the majority
easy to handle when compared to animals of which are related to industrial
and plants. They require lesser space and production. The growth of microorganisms
serve as more cost-effective sources. in a liquid broth is referred to as submerged
2) Microorganisms can be easily fermentation. The method often
manipulated using genetic engineering or necessitates a large amount of oxygen.
other means. They can be subjected to When the microorganisms interact with the
strain improvement, mutations and other nutrients in the broth, they break them
such changes by which the production of α- down, resulting in the formation of
Amylase can be optimized. Also, the enzymes. In the fermentation broth,
microorganisms can be tailored to cater to bioactive compounds are secreted.
the needs of growing industries and to Solid State fermentation:
obtain enzymes with desired characteristics Solid State Fermentation (SSF) is a
like thermostability for example. fermentation process used by a variety of
Thermostable α-Amylases are desired as industries, including pharmaceuticals, food,
they minimize contamination risk and and textiles, to manufacture microorganism
reduce reaction time, thus saving metabolites using a solid support rather
considerable amount of energy. Also, when

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than a liquid medium. It's known as approximately 54 kDa. Saliva amylases

microbe growth in the absence of a free- hold 40% to 50% of complete protein
flowing aqueous process. For the saliva and most of the enzyme associated
processing of value-added products such as with parotid hunger (80% of number) [7].
antibiotics, single-cell protein, PUFAs, Calcium metalloenzyme containing
enzymes, organic acids, biopesticides, hydrolyzes α-1,4 links starch in glucose and
biofuel, and aromas, the SSF is a viable maltose. It is known to be involved mainly
alternative to submerged fermentation. in the initiation of digestion starch in the
Grain brans, de-oiled oil seed cakes, and oral cavity. However, Salivary α-amylase
other materials are among the materials has also been shown to have an important
used as a support. Initially, fungi were function in viral interactions [9].
primarily used in this form of fermentation Microbial sources:
(as these microorganisms were considered In addition to the widespread distribution of
to be very optimally active in very low enzyme, microbic sources, particularly
water activity). Later, a variety of bacterial fungous similarly bacterium enzyme, that is
species and yeasts were used in the employed in industrial production owing to
fermentation process. such advantages as prices efficiency,
Organism used in SSF: consistency, period and easy method
SSF can also be made with bacteria and modification and potency [10]. Fungal
yeasts, which require a higher moisture amylases are widely utilized in the
content for efficient fermentation, but with preparation of oriental food [12]. In
a lower yield. between bacteria, Bacillus sp. is widely
SOURCES OF AMYLASE: utilized in the assembly of volatile α-
Animal sources: amylase to fulfill business requirements. B.
Ptyalin, salivary α-amylase (α-1,4-α-D- subtilis, B. stearothermophilus, B.
glucan-4-glucanohydrolase; EC 3.2.1.1) licheniformis
one of the most important enzymes in the a, and B. amyloliquefaciens are renowned
saliva. The enzyme was expressed by have been sensible producers of α-amylase
tithing by Leuchs in 1831 [7]. It consists of and these are widely utilized in business
two families of isoenzymes, one of which is production of a catalyst for varied
glycosylated and the other does not contain applications [13]. Similarly, threadlike
carbohydrates. The molecular weight of the fungi have widely used the production of
glycosylated form is approximately amylases for hundreds of years. As these
57 kDa,that of the non-glycosylated form is fungi are removed to be the leading

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producers of foreign proteins, they are [17]. Remains of true bacteria

extremely exploited within the production stearothermophilus and B. Amylolyticus
of assorted enzymes including as well. secreted high alpha enzyme in a
PARAMETERS AFFECTING neighborhood infused with I Chronicles
ENZYME ACTIVITY organic compound, 0.5% yeast extract, and
Carbon sources: zero.5% malt6ose below intense vibration
Carbon sources such as galactose, conditions [20]. L-asparagine was is
glycogen, and Inulin have been reported as rumored to be one of the foremost
suitable substrates for amylase production promising sources of N sources of alpha
by B. licheniformis and Bacillus [11]. enzyme production by Thermomyces
Starch and glycerol were known to increase lanuginosus [19].
enzyme production in B. subtilis IMG22, Temperature
Bacillus sp, PS-7, and Bacillus sp. 1-3 [18]. The effect of heat on amylase production is
Soluble starch has been found as a leading related to body growth [3]. Therefore, the
substrate production of alpha-amylase maximum temperature depends on whether
by B. stearothermophilus [19]. Bacillus sp. the culture is mesophilic or thermophilic. In
it was noted that the high given raw starch the middle of the mold is a lot of amylase
digests amylase in the area containing production studies were performed with
lactose (1%) and yeast extraction (15%). mesophilic mold within a temperature of
Agricultural waste is used in both liquid 25-37°C [22]. A raw amylase-damaging
and solid fermentation to reduce the cost of starch was made by Aspergillus ficum at
fermentation media. Waste contains carbon 30°C by Hayashida in 1986. Amylase
and nitrogen resources needed for physical production in High levels has been reported
growth and transformation. These nutrient between 50-55°C by fungalophilic fungal
sources include orange peel, peanuts cultures such as Talaromyces Emersonii
for peers’ starch, potatoes, corn, tapioca, etc. [21].
wheat, and rice as flour [16]. pH:
Nitrogen sources: pH is one of the important factors that
Soybean is a wonderful supply of N for determine the growth and morphology of
alpha enzyme by true bacteria sp. 1- microorganisms as they are sensitive to the
3. Tanyildizi et al rumored that organic concentration of hydrogen ions in
compounds increased accelerator activity between[19]. Previous studies have shown
whereas the discharge of yeast failed to that mold is needed acidic pH and bacteria
show a sway on alpha enzyme production require a neutral pH to grow properly. pH

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IJBPAS, April, Special Issue, 2021, 10(4)
Upadhyay D et al Review Article

is known for incorporating and secreting attempt to produce α-Amylase from A.

pH alpha-amylase [24]. Aspergillus fungus oryzae using coconut oil cake as a
sp. as A. oryzae, A. ficuum, and A. substrate, the main activity was found to be
Niger have been found to provide a high 1752 U / gds which resulted in 72 h
yield of alpha-amylase at a pH of 5.0 to 6.0 incubation [32]. The positive α-Amylase
in SmF [23] Alpha-amylase species that activity of 62 470 U / g was obtained after
produce yeast such 72 hours of fermentation during the
as Cerevisiae and Sekyere have shown high production of Bacillus amyloliquefaciens
enzyme production at pH 5.0 [22]. [34]
DURATION OF FERMENTATION APPLICATION
This is an important factor in the Amylase has been used in various
fermentation process. If the process is done industries. Like Alcohol industries,
shorter than the time limit the maximum Chocolate industries, Paper industries,
yield cannot be obtained. Enzyme activity Bread Industries, Textile Industries, Biofuel
increases with increasing incubation time Industries, Detergent industries [35].
until you reach the best time. In most cases, CONCLUSION
enzyme production begins to decline as the Amylase is known as one of the oldest
incubation period increases further. This enzymes. And it was discovered by
can be caused by nutrient depletion in the Kirchhoff, 1815. It has been divided into
middle [31, 32] or the release of toxic three types: α, ß, and Γ amylase. Mainly
substances. Bacillus subtilis, a well-known amylase is used in industries like the paper
producer of alpha amylase was studied to industry, chocolate industry, textile
compare between different hours of industry, bread industry, the alcohol
fermentation and studies have shown a high industry and Bio-fuel industry, Detergent
yield of alpha amylase after 48 hours of industry, etc.
fermentation [33]. The duration of the ACKNOWLEDGEMENT
incubation period was compared with the It's our privilege and honour to express our
amylase product from Penicillium sincerest gratitude to the Parul University,
fellutanum separated from the mangrove Vadodara, Gujarat for providing me all the
rhizosphere soil. The culture when necessary support and facilities including
incubated at 96 h, showed a significant state of the art infrastructural facilities with
activity of 136 U / ml. This was a double advanced technological scientific
increase in activity compared to a 24 h laboratories and everything else that was
incubation (68 ± 2.3 U / ml) [12]. In an required to carry out this work.

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Upadhyay D et al Review Article

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