Isolation, Screening and Identification of Protease Producing Bacteria from Fish Sauce
Main Article Content
Abstract
Proteolytic microorganisms in fish sauce are believed to play an important role in fish sauce fermentation. For better understanding their role in fish sauce fermentation and applying them back to the process, in this study the isolation and screening of protease-producing strain from fish sauce was investigated. The results showed that the proteolytic bacteria could be isolated on skim milk salt agar at salt concentration 5%, 7.5% and 10% but not at 20% or 25%. 33 isolates with halo was selected and was screened further by spot inoculation on skim milk salt agar at 5% and 10% NaCl and by enzyme diffusion method on skim milk agar at salt concentration 10%. All 9 isolates selected by high halo’s diameter D-d were gram positive spore forming rod and were identified as Virgibacillus halodenitrificants by 16s rDNA analysis with more than 99.2% homology. The strain Virgibacillus halodenitrificants CH201 possessed highest halo diameter 12.33 mm by enzyme diffusion method, following by CH322 9.33 mm. These two strains showed different profile of sugar utilization on API kit CH50.
Keywords
fish sauce, Virgibacillus halodenitrificants, isolation, protease
Article Details
References
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[10] Le Van Viet Man, Tran Thi Anh Tuyet; Characterization of protease from Aspergillus oryzae surface culture and application in fish sauce processing; Tạp chí phát triển KH&CN. 9 (2006) 53-58.
[11] J. Yongsawatdigul, S. Rodtong, and N. Raksakulthai; Acceleration of Thai Fish Sauce Fermentation Using Proteinases and Bacterial Starter Cultures; Journal of Food Science. 72 (2007) M382-M390.
[2] S. Sinsuwan, S. Rodtong, and J. Yongsawatdigul; Evidence of cell-associated proteinases from Virgibacillus sp. SK33 isolated from fish sauce fermentation; J Food Sci. 76 (2011) C413-9.
[3] S. Sinsuwan, S. Rodtong, and J. Yongsawatdigul; Hydrolytic activity of Virgibacillus sp. SK37, a starter culture of fish sauce fermentation, and its cell-bound proteinases; World J Microbiol Biotechnol. 28 (2012) 2651-9.
[4] T. C. M, M. T. J., S. P., and G. W. D; Isolation and characterization of an extremely halophilic archaebacterium from traditionally fermented Thai fish sauce (nam pla); Letters in Applied Microbiology. 14 (1992) 111-114.
[5] S. Chaiyanan, S. Chaiyanan, T. Maugel, A. Huq, F. T. Robb, and R. R. Colwell; Polyphasic taxonomy of a novel Halobacillus, Halobacillus thailandensis sp. nov. isolated from fish sauce; Systematic and Applied Microbiology. 22 (1999) 360-365.
[6] K. Hiraga, Y. Nishikata, S. Namwong, S. Tanasupawat, K. Takada, and K. Oda; Purification and Characterization of Serine Proteinase from a Halophilic Bacterium Filobacillus sp. RF2-5; Bioscience, Biotechnology, and Biochemistry. 69 (2005) 38-44.
[7] N. Udomsil, S. Rodtong, S. Tanasupawat, and J. Yongsawatdigul; Improvement of Fish Sauce Quality by Strain CMC5-3-1: A Novel Species of Staphylococcus sp.; Journal of Food Science. 80 (2015) M3215-M3202.
[8] W. J. Kim and S. M. Kim; Purification and characterization of Bacillus subtilis JM-3 protease from anchovy sauce; Journal of Food Biochemistry. 29 (2005) 591-610.
[9] N. V. Lâm, N. P. Nhuê, and T. T. Ngọc; Phân lập và nghiên cứu vi khuẩn ủ muối, ủ kiểm sinh enzyme protease và bước đầu thử nghiệm để sản xuất nước mắm ngắn ngày; Tạp chí khoa học và phát triển. 9 (2011) 352-463.
[10] Le Van Viet Man, Tran Thi Anh Tuyet; Characterization of protease from Aspergillus oryzae surface culture and application in fish sauce processing; Tạp chí phát triển KH&CN. 9 (2006) 53-58.
[11] J. Yongsawatdigul, S. Rodtong, and N. Raksakulthai; Acceleration of Thai Fish Sauce Fermentation Using Proteinases and Bacterial Starter Cultures; Journal of Food Science. 72 (2007) M382-M390.