Enviromental Microbiology

Two fractions showing proteolytic enzymes have been obtained from the latex of Synadenium grantii Hook, ‘f’, using gel-filtration and anion-exchange chromatographic techniques. Both these proteases have the same molecular mass of 76±2 kDa each. They exhibit maximal activity at pH 7.0 and at a temperature of 60 °C. They display stability over a pH range from 5–10 and are also highly thermostable. Irreversible inhibition by PMSF indicates that they are serine proteases. In addition, histidine residues also appear to play an important role in catalysis as evidenced by inhibition with DEPC. They also exhibit similarity with respect to pH and temperature optima, kinetic properties and thermal stability.

Plant residuals comprise the natural habitat of the plant pathogen; therefore, attention is currently focusing on biological-based bioprocessing of biomass residuals into benefit substances. The current study focused on the biodegradation of peanut plant residual (PNR) into citric acid (CA) through a mathematical modeling strategy. Novel endophytic Trichoderma longibrachiatum WKA55 (GenBank accession number: MZ014020.1), having lytic (cellulase, protease, and polygalacturonase) activity, and tricalcium phosphate (TCP) solubilization ability were isolated from peanut seeds and used during the fermentation process. As reported by HPLC, the maximum CA (5505.1 μg/g PNR) was obtained after 9 days in the presence of 15.49 mg TCP, and 15.68 mg glucose. GC-MS analysis showed other bioactive metabolites in the filtrate of the fermented PNR. Practically, the crude product (40%) fully inhibited (100%) the growth and spore germination of three mycotoxinogenic fungi. On peanuts, it improved the seed germination (91%), seedling features, and vigor index (70.45%) with a reduction of abnormal seedlings (9.33%). The current study presents the fundamentals for large-scale production in the industry for the sustainable development of PNR biomass as a natural source of bioactive metabolites, and safe consumption of lignocellulosic-proteinaceous biomass, as well. T. longibrachiatum WKA55 was also introduced as a novel CA producer specified on PNR. Application of the resulting metabolite is encouraged on a large scale.

ABSTRAK Telah dilakukan penelitian " Isolasi Dan Karakterisasi Bakteri Proteolitik Dari Saluran Pembuangan Limbah Industri Tahu ". Penelitian ini bertujuan untuk mendapatkan isolat bakteri proteolitik dari saluran pembuangan limbah industri tahu dan mengetahui karakter serta kemampuan aktivitas proteolitik sebagai penghasil enzim protease. Isolasi bakteri proteolitik menggunakan medium SMA (Skim Milk Agar). Pengamatan karakter isolat bakteri proteolitik terlebih dahulu dilakukan, seleksi isolat bakteri, dan selanjutnya pengamatan morfologi koloni, pengecatan gram, dan pewarnaan spora. Hasil seleksi isolat bakteri proteolitik menunjukkan terdapatnya 7 isolat bakteri yang dapat menghasilkan zona halo dan terdapat dua isolat bakteri tersebut yang memiliki nilai Indeks Proteolitik tertinggi. Pengamatan morfologi bertujuan untuk menunjukkan adanya variasi warna, bentuk, tepi dan elevasi dari setiap isolat. Hasil pengecatan mikroskopis menunjukkan terdapat 2 isolat bakteri bersifat gram positif dan memiliki bentuk sel berbentuk batang dan tidak memiliki endospora. Hasil pertumbuhan bakteri menunjukkan 2 isolat bakteri memiliki fase logaritmik dengan waktu generasi yang berbeda yang masing-masing isolat A1 = 240 menit dan isolat A4 = 168 menit. Pengujian aktivitas proteolitik menunjukkan 2 isolat bakteri menghasilkan nilai aktivitas enzim berbeda yaitu isolat A1 sebesar 0,0044 U/mL dan isolat bakteri A4 sebesar 0,1498 U/mL. The research have done about " Isolation and Characterization of Proteolytic Bacteria From Industrial Waste Sewer of Tofu ". The research has objective to obtain proteolytic bacteria from sewage with industrial of tofu and know that character along with ability of proteolytic activity as produce protease enzymes. Isolation of proteolytic bacteria using by Skim Milk Agar medium. Observations character of proteolytic bacteria, beforehand do the selection of bacterial isolats, and then observing of colony morphology, gram staining, and spores staining. The result of Selection proteolytic bacteria isolats showed seven isolats that produce halo zone and there are two bacterial isolats that have the highest value proteolytic index. Morphological observation aims to show the variation of color, shape, edge and elevation of each isolat. The results of microscopic staining showed that there were two isolatd bacteria are gram-positive and has has the shape of stem cells and didn't had endospores. The results showed two isolats have logarithmic phase and a different generation on the bacterial growth that is A1 = 240 minute and A4 = 168 minute. Proteolytic activity test showed two isolats different value of produce enzyme activity that is A1 as much as 0,0044 U/mL and A4 isolats much as 0,1498 U/mL.

Proteases are enzymes used in industries such the production and processing of detergents, food, leather, and silk. The aim of this study was to isolate and identify protease-producing bacteria from a sludge disposal site and from sediments. Soil samples were collected separately from the selected area. Samples weighing 1 g were serially diluted and spread onto skim milk agar. A total of 16 bacteria species were isolated from the study samples. Four bacterial isolates showed high proteolytic activity and were selected for enzymatic study based on their zone of proteolysis. The isolates were identified based on biochemical tests. The results indicated that the isolated bacteria were E. coli (99.69%), Pseudomonas putrefaciens (Shewanella putrefaciens) (91.61%), Bacillus carboniphilus (92.78%), and Lysinibacillus sphaericus (98.4%). The crude protease enzymes produced by these bacterial isolates showed promising results for application in dehairing and destaining as detergent additives. Bacillus carboniphilus showed the best level of activity and was selected as the most potent protease-producing bacteria for both dehairing and destaining ability. Soils from sludge disposal sites and sediments from around tannery wastes could be good sources from which to isolate alkaline protease-producing bacteria.

Problem statement: Nitrate and phosphate are two major pollutants due to anthropogenic activity like excessive use of fertilizers in agriculture. Their contamination has emerged as a global problem and its potential threat is marked on the environmental sustainance as well as on the public health. Approach: The objective of the current study is to isolate efficient nitrate and phosphate removing microbes from various environmental sites that have been selected on the basis of the nature of polutants received by them and their water quality assessment. These well characterized isolates could in future be used for the remediation of waste water. 30 different sites were screened using culture based method. The nitrate and phosphate removing abilities of the microbes were checked in enriched medium (Himedia M439) after 16 h of incubation at 37°C. Results: 7 efficient isolates were obtained from rhizosphere of Water lily, Marine beaches, Paddy field and Raw sewage canal. The highest nitrate removal (88.3%) was shown by isolate (WBUNB009) from raw sewage canal and the highest phosphate removal (82.9%) was shown by isolate (WBUNB004) from rhizosphere of Water lily. Morphologically all the isolates were gram positive bacilli as reconfirmed by environmental scanning electron microscopy. Biochemically as well as physiologically they differ from each other. Conclusion/Recommendation: This study leads to the isolation of efficient nitrate and phosphate removers from environmental origin. The phosphate removing efficiency is much higher than the type strain under identical condition. These native microbes might be responsible for maintaining the phosphate and nitrate levels at the 30 sites investigated inspite of the received pollution load. These isolates could be the potential bioremedial agents for other sites with high nitrate and phosphate contamination level.

Objective – To identify the spatial and seasonal variation of microorganisms associated with Zungaro jahu cultivation semi-intensive.
Methods – Water samples were collected along a longitudinal axis in a fish farming Conceição das Alagoas county, Minas Gerais - Brazil,
and seeded in different culture medium. Results – Ten bacteria genus were isolated and identified 11 of microorganisms and 3 species of
Gram-negative and Gram-positive bacteria, respectively was obtained in the five collected points analyzed. The seasonal variation analysis
indicated a reduction of taxonomic categories in July, must be directly associated with the low temperatures. Conclusion – The analysis showed
that space variation of the tank which receives water from the whole farm-fish has a higher microbial diversity; probably due to the addition
of organic matter used in fish feed

The use of bioagents is risk free and the best
alternative to sustain plant protection. Rhizoctonia solani
affect the seed and seedling health of chickpea by causing
seed and seedling rot. It showed that all the Trichoderma
isolates inhibited significantly the growth of the pathogen
and it ranged from 41.4 (T. viridi 1) to 52.8 (T. viridi 2).
Germination per cent ranged from 74 (T. viridi 4) to 90.3
(PDBTCH 10) was when treated with bio-agent where as in
pathogen check it was only 32.6%. Maximum vigor index
(2239.4) was recorded in T. harzianum (PDBCTH 10)
where as in pathogen check it was 169.4 only. All bioagents
treatment recorded significantly less disease incidence
as compared to pathogen check. Minimum (13.3%)
pre emergence mortality was recorded in seeds treated with
T. viridi 2 Lowest post emergence mortality was recorded
in T. viridi 1 (4.8%). In remaining bioagents treatments the
post emergence mortality ranged from 16.7 to 27.9%.
The results obtained clearly indicated that the efficacy of
different isolates of same species also showed variation
when tested against the same pathogen, therefore, it may be
concluded that before application in the field the potentiality
of every strain of the bioagents should be tested
against target pathogen

In the present study, an attempt is made to evaluate the kinetics of biological phosphate removal using a bacterial consortium of activated sludge, as well as screening for dominant polyphosphates accumulating bacteria. The results showed an efficient phosphate uptake (P < 0.001) of the consortium, with rates related to the initial concentration of both phosphate and carbon sources. Short chain volatile fatty acids presented the suitable substrates for enhanced biological phosphorus removal, of which maximum yield reached 99.23% and 78.51% in basal salt medium supplemented with 0.5% of sodium acetate and lactate respectively. Fifteen phosphate-accumulating bacteria were isolated from the activated sludge and only four isolates were selected and characterized as Pseudomonas aeruginosa AS1, Moraxella lacunata AS2, Acinetobater junii AS3 and Alcaligenes denitrificans AS4. The highest efficiency of phosphate uptake using pure culture was achieved with Ac. junii AS3 (83.36) followed by P. aeruginosa AS1 (81.78%), Al. denitrificans AS4 (76.72%), and M. lacunata AS2 with 50.6%.

The objective of this investigation was to assess the sanitary status of groundwater to ensure hygienic
requirement water standards. Pathogens in water may originate from animals and anthropogenic waste. In
a semi-rural area microorganisms are significant indicators connected with human activities in the underground water. Notably, our results showed that a more efficient diagnostic group to determine the level of
contamination was proteolytic bacteria (p<0.01). Results showed that pathogenic bacteria such as Shigella
sp., Salmonella sp., Escherichia coli, and total coliforms were not detected. The highest number of Staphylococcus sp. and fungal spores were found in the spring. Considerable additional research is needed to
appropriately measure microbial communities in underground water.