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Characterization of Protease Produced by Bacillus amyloliquefaciens HTP-8 Isolated from Korean Fermented Anchovy Sauce.  

임형택 (경남대학교 생명과학부)
정순경 (양산대학 식품가공제과제빵과)
김기남 (경남대학교 생명과학부)
하정욱 (경남대학교 생명과학부)
백현동 (경남대학교 생명과학부)
Publication Information
Microbiology and Biotechnology Letters / v.30, no.1, 2002 , pp. 26-32 More about this Journal
Abstract
For commercial production of Korean fermented anchovy sauce through rapid fermentation, a bacterial strain which showed the high protease activity was isolated from a commercially fermented anchovy sauce. The isolate was Bacillus amyloliquefaciens, and named as B. amyloliquefaciens HTP-8. The incubation temperature, initial pH, and cultivation time for optimal production of protease by B. amyloliquefaciens HTP-8 were $30^{\circ}C$, 7.0, and 3 days, respectively. In jar fermenter, B. amyloliquefaciens HTP-8 showed higher protease activity when grown at pH 7.0. The protease was partially purified by 80% ammonium sulfate precipitation and CM-Sephadex C-50 ion exchange chromatography. The partially purified enzyme had specific activity of 103.3 units/mg, yield of 0.4%, and purification fold of 43.0. The optimal pH and temperature for the protease activity were 10.0 and $50^{\circ}C$, respectively. The protease was relatively stable at the pH range of 7.0~12.0 and at the temperatures below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $Ag^{＋}$ /, $Ba^{2＋}$ and selectively inhibited by PMSF, suggesting that it is a serine protease.
Keywords
Korean fermented anchovy sauce; Protease; Bacillus amyloliquefaciens;
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1 Hagihara, B., H. Matrubara, M. Nakai, and K. Okunuki. 1958. Crystalline bacterial proteinase. I. Preparation of crystalline proteinase of Bacillus subtilis. J Biochem. 45: 185194.   DOI
2 Horikoshi, K. and T. Akiba. 1982. Alkalophilic Microorganisms. pp 93-101. Scientific Societies Press, Tokyo.
3 Lee, E. G., E. H. Park, and H. H. Hyun. 2000. Purification and characterization of two alkaline proteases produced by Pseudomonas sp. BK7. J Microbiol. Biotechnol. 10: 677684.
4 Lim, S. I. and J. Y Yoo. 1999. Purification and characteristics of protease produced by Syncephalastrum racemosum PDA 132-2 from Korean traditional meju. J Korean Soc. Food Sci. Nutr. 28: 1010-1016.
5 Kobayashi, T., A. Ogasawara, S. Ito, and M. Saitoh. 1985. Purification and some properties of alkaline proteinase produced by Pseudomonas maltophilia. Agric. Bioi. Chem. 49: 693-698.
6 Yeum, D. M., T. G. Lee, J. R. Do, O. K Kim, Y. B. Park, S. B. Kim, and Y. H. Park. 1993. Characteristics of angiotensinI converting enzyme inhibitors derived from fermented fish product. 2. Characteristics of angiotensin-I converting enzyme inhibitors of fish sauce prepared from sardine, Sardinops melanosticta. Bull Korean Fish. Soc. 26: 416-423.
7 Hwang, S. Y 1995. Purification and characterization of an extracellular serine protease from Bacillus sp. strain KUN17. Kor. J Appl. Microbio!. Biotechnol. 23: 53-59.
8 Lim, S. I. 2000. Purification and characterization of protease produced by Aspergillus wentti isolated from Korean traditional meju. Korean J. Food Sci. Techno!. 32: 161-167.
9 Lim, S. I., H. K. Kim, and J. Y. Yoo. 2000. Characteristics of protease produced by Bacillus subtilis PCA 20-3 isolated from Korean traditional meju. Korean J Food Sci. Technol. 32: 154-160.
10 Johnvesly, B. and R. N. Gajanan. 2001. Production of bleach-stable and halo-tolerant alkaline protease by an alkalophilic Bacillus pumilus ill 05 isolated from cement industry effluents. J Microbiol. Biotechnol. 11: 558-563.
11 Kim, S. J., D. B. Shin, and B. H. Ryu. 1993. Continuous rapid fermentation of sardine soy sauce by using column type reactor packed immobilized yeast cells. Korean J. Food Sci. Technol. 25: 154-159.
12 Ok, M., M. S. Kim, W. S. Seo, J. Y. Cha, and Y. S. Cho. 2000. Characterization of extracellular protease of Bacillus sp. WRD-1 isolated from soil. Kor. J Appl. Microbiol. Biotechnol. 28: 329-333.
13 Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall. 1951. Protein measurement with the folin phenol reagent. J Bioi. Chem. 193: 265-275.
14 Sakka, K., K. Shimada, and K. I. Matsushima. 1985. Purification and some properties of serine proteinase from a mutant of Aspergillus niger. J Ferment. Technol. 63: 479483.
15 Lee, E. H., T. H. Lee, J. S. Kim, and C. B. Ahn, 1989. Processing and taste compounds of the fish sauce from skipjack scrap. Bull. Korean Fish. Soc. 22: 25-35.
16 Lee, E. H., H. S. Park, C. B. Ahn, and G. C. Hwang. 1986. Preparation of fish sauce from mackerel scrap. J Korean Soc. Food Nutr. 15: 201-206.
17 Yun, S. w., K P. Lee, J. H. Yu, C. S. Shin, and D. H. Oh. 1989. Purification and properties of alkaline protease from Streptomyces sp, YSA-130. Kor. J Appl. Microbiol. Biotechnol. 17: 358-364.
18 Lee, E. H., C. B. Ahn, J. S. Kim, C. W. Lim, S. W. Lee, and Y. A. Choi. 1988. Processing and taste compounds of fish sauces from filefish scrap. J Korean Soc. Food Nutr. 17: 326-335.
19 Ryu, B. H., S. J. Kim, and D. B. Shin. 1992. Lactic acid, ethylalcohol and 4-ethylguaiacol contents of rapid fermentation of sardine soy sauce prepared by using immobilized whole cells. KoreanJ Food Sci. Technol. 24: 456-462.