Comparative study of proteolytic activity of protease-producing bacteria isolated from thua nao (original) (raw)
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Journal of Microbiology, 2011
The present study was investigated to optimize and partially purify the proteases produced by the food borne bacterial strains. Methodology and Results: Four bacterial strains such as Bacillus cereus, Proteus vulgaris, P. mirabilis and Enterobacter aerogenes were isolated from food wastes. These strains were individually inoculated in to the formulated culture media supplied with three different concentrations (1:1 to 1:3) of raw milk as major substrate. Among the concentrations, 1:2 ratio of substrate supplied medium showed maximum (0.133 to 8.000 IU/mL) protease production by all the tested organisms. After optimization, the organisms were tested for protease production at various pH (3 to 9), and temperature (30 to 80 °C). The result showed that all the organisms were capable of producing maximum protease at pH 6 (8.533 to 10.133 IU/mL) and at 50 °C (8.666 to 10.666 IU/mL). The crude enzymes produced by the tested organisms were individually purified by two different methods viz sodium alginate and ammonium sulphate-butanol methods. The purity of the protease determined in these two methods was ranged between 3.24 to 5.44 I and 3.13 to 5.55 IU/mL respectively. The partially purified enzymes were further analysed through SDS-PAGE; accordingly the molecular weight of protein produced by the test organisms was determined in between 49.44 and 50.98 kDa. Conclusion, significance and impact of study: Among the tested strains P. vulgaris was identified as the major protease producer in optimized culture condition of 50 o C and pH6. The molecular mass of the partially purified protease of P. vulgaris was 50.32 KDa. Further research on optimization of other fermentation parameters using statistical tools with P. vulgaris is needed to scale up the process.
Study of Protease Producing Bacteria and their Enzymatic Activity at Different Parameters
International Research Journal of Engineering & Technology, 2019
Bacterial alkaline proteases are among the important hydrolytic enzyme and have been used extensively since the advent of enzymology. Bacterial extracellular alkaline proteases are of great importance due to its wide spectrum applications in detergent industries, bioremediation, food industries, and leather processing and bio-film degradradation. The Optimization of the time of incubation, the effect of temperature, the effect of pH, the effect of different carbon source and effect of different nitrogen source and they were found to be 120hrs, 12, 170Unit/ml for galactose, and 70 Unit/ml for ammonium sulfate respectively for the production of protease and also optimize the parameters on enzyme activity they are temperature, pH, 1% casein substrate, and the different volume of the crude enzymes and the results was 370C,11, 3.5ml, 60 Unit/ml for 2 ml of crude enzymes respectively.
Energy Reports, 2020
The objective of the study to determine the factors affecting the protease content generated during soybean fermentation by Bacillus subtilis. In this study, four factors were investigated during soybean fermentation: (1) pH, (2) fermentation temperature, (3) fermentation time and bacterial density, and (4) added glucose content on steamed soybean. The experimental results showed that at 33 • C temperature, 48 h fermentation time, bacterial density 10 4 CFU, pH 7, and supplementation of 1 g glucose into 50 g steamed soybean, Bacillus subtilis fermented soybean (another name is natto) had the highest protease activity. This suggests that the fermentation conditions in the study are consistent with the growth of Bacillus subtilis to produce proteases. Moreover, the optimization of fermentation conditions contributes to reducing the energy consumption to obtain the desired product, by increasing the yield in reaction. Therefore, this study is important at an industrial level, while contributing to the rational use of energy. c
Pure and Applied Biology
The purpose of this study was production of protease enzyme by isolated bacteria and evaluation of their potential for various industrial applications. Two strains AN16 and AN13 exhibiting maximum proteolytic activity were used for further studies. The activity of crude enzymes was completely inhibited by phenyl-methyl-sulphobnyl fluoride (PMSF) and ethylene diamine tetraacetic acid (EDTA) had no such negative effect on activity. Enzymes were relatively stable in presence of surface active agents such as Triton X-100, Tween-80, Tween-20 and oxidizing agent H₂O₂. The enzymes exhibited maximum activity within pH range of 7-9. The optimum temperature was recorded to be 37⁰C. The relative activity of isolate AN13 protease in presence of metal ions Ca2+,
ScienceAsia
Eighty-two bacillus strains were isolated from Thai fermented soybeans (thua nao), of which thirty-nine were identified as Bacillus subtilis. Crude proteins from these B. subtilis strains were investigated for their proteolytic activity. When tested on skim milk agar, the crude proteins of B. subtilis strain 38 exhibited the highest proteolytic activity (a clear zone with an average area of 480 mm 2). Optimal pH and temperature of the proteases from B. subtilis strain 38 were found to be at 6.5 and 47 °C , respectively. The proteases were unstable and rapidly decreased in activity when heated at 60 °C. Various protease inhibitors were tried, only 1, 10-phenanthroline decreased the enzyme activity indicating the presence of metalloproteases. In addition, we attempted to produce fermented soybeans using the B. subtilis strain 38 as a starter culture.
Microbiology and Industrial Biotechnology of Food-Grade Proteases: A Perspective
2006
Summary Recent developments in industrial biotechnology have resulted in the exploitation ofnew and undiscovered microorganisms and the devising of improved methods for en-zyme production, which have led to increased yields of the enzyme, thus making a viableindustrial process feasible. This review tracks the developments in the field of acidic andneutral protease production with regard to the producers, methods of production andtheir improvement, the product and its applications. Key words : neutral protease, acidic protease, fermentation, genetic engineering, purifica-tion, enzyme assay Introduction Proteases are one of the industrially most importantenzymes. These proteolytic (protein digesting) biocata-lysts have been in use for many centuries, at first in thedairy industry as milk-clotting agents (rennet) for themanufacture of cheese. Proteases are enzymes that cata-lyse hydrolytic reactions in which protein molecules aredegraded to peptides and amino acids. These constitutea v...
Biotechnological Perspectives of Microbial Proteases
The review briefly elucidates the importance of proteases in living organisms and their wide range of potential applications in the vast areas of research and biotechnology. The important features of the proteases are also exploited in a number of ways and can be used to serve various applications in different industries. Apart from their natural potential of performing various important operations in living tissues, various microbial proteases have potential applications in a number of industries. In cellular environment, proteases are involved in the breakdown of the proteins' peptide bonds and transform them into smaller fragments of amino acids and peptides which are prerequisite for the differentiation and cellular growth. Proteases have also vast applications in a range of industrial procedures such as food, pharmaceutical, dairy and detergent. Microbial proteases have dominated roles in the industrial sectors. Microbial proteases are exploited for their characteristic feature of hydrolyzing the protein and the rest of the components of wheat and soy beans in the production of soy sauce. The production of proteases can be enhanced via substantiated fermentation methods. The variation in the composition of growth media such as changes in carbon and nitrogen ratio and some other features affecting microbial growth are significant in the evaluating the fermentation procedures. The production of microbial proteases is advantageous because they can be generated rapidly, their production is cost effective and the manipulation of microbial enzymes is quite easy. Proteolytic enzymes can be produced by either submerged fermentation (SmF) or solid state fermentation (SSF). But the latter is far more advantageous because it direct towards many potential benefits for the protease production. The review mainly focuses on the microbial protease production, their functional and structural aspects and the application of these proteolytic enzymes in different industries.
INFLUENCE OF CULTURE CONDITIONS ON PRODUCTION AND ACTIVITY OF PROTEASE FROM BACILLUS SUBTILIS BS1
Bacillus subtilis BS1 was used in the present study for the production of protease. For protease production optimum pH and temperature were found to be 11 and 50˚C, respectively. Soybean (197 PU/mg) and casein (168 PU/mg) proved as the best substrates for the production of enzyme. Maximum production of protease (126 and 121 PU/mg) was shown in 1.5 and 4.5% of Sodium chloride (NaCl) concentration, respectively. Maximum activity was observed at pH 9 at 90ºC, in crude extract, after 20 minutes of incubation. EDTA slightly enhanced proteolytic activity, whereas, Na, K, Ca, Li, Mg, Cu and Fe inhibited the activity of protease. Due to maximum production of protease in the presence of cheaper, low concentrations of substrate and stability at alkaline pH, high temperature and salt-tolerance, these characteristics makes the strain and its enzymes usefull in different industries.
Journal of Agricultural and Food Chemistry, 2003
Bacillus amyloliquefaciens FSE-68 isolated from meju, a Korean soybean fermentation starter, was identified on the basis of biophysical tests and 16S rRNA gene sequence. A neutral metalloprotease (NPR68) and an alkaline serine-protease (APR68) were purified by ammonium sulfate precipitation and cation exchange chromatography and identified on the basis of their activities at different pH values and the selective protease inhibitors. The molecular weights of NPR68 and APR68 measured with ESI-MS were 32743 (( 0.8) and 27443 (( 0.5) Da, respectively. Against oxidized insulin chains, the NPR68 has a cleavage preference at the site where leucine is located as a P1′ residue followed by phenylalanine, and the APR68 has broad specificity and favors leucine at the P1 site. These results indicate that the proteases are natural variants of subtilisin and bacillolysin.