A novel alkaline serine protease from bacillus amyloliquefaciens strain S1-13
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Published:
Jul 15, 2016
Keywords:
Bacillus amyloliquefaciens 16 rRNA gene alkaline serine protease metal-dependent enzyme and LC/MS-MS
Main Article Content
Abstract
Bacillus sp. strain S1-13 was identified as Bacillus amyloliquefaciens with 16s rRNA gene (Accession number: JX441363). The strain S1-13 was expressed and secreted alkaline serine protease (called ASP1-13) when growth in nutrient broth containing with 1% skim milk. ASP1-13 was partial purified with a specific activity of 1,324 U/mg and 2% yield. The molecular weight and isoelectric point of ASP1-13 was determined about 40 kDa and 8, respectively. It was indicated as alkaline serine protease with a broad range of activity at alkaline condition (pH 7-12) and completely inhibited with serine protease inhibitor. ASP1-13 was also active in high temperature (50-60°C) and stabilizes with broad range of pH (5-12), surfactant, oxidant, reducing agent and organic solvent. Finally, the partial amino acid sequence from LC/MS-MS was confirmed similarity with sequence of neutral protease precursor from Bacillus subtilis.
Article Details
How to Cite
Yingchutrakul, Y., Roytrakul, S., Chukeatirote, E., & Waratrujiwong, T. (2016). A novel alkaline serine protease from bacillus amyloliquefaciens strain S1-13. Asia-Pacific Journal of Science and Technology, 21(2), 127–139. https://doi.org/10.14456/kkurj.2016.18
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Research Articles
References
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[2] Garcia-carreno FL. Protease inhibition in theory and practice. Biotechnology Education. 1992;3(4):145-50.
[3] Gupta R, Beg QK, Lorenz P. Bacterial alkaline proteases: molecular approaches and industrial applications. Applied microbiology and biotechnology. 2002 Jun;59(1):15-32.doi:10.1007/ s00253-002-0975-y.
[4] Ray A. Protease Enzyme Potential Industrial Scope. International Journal of Technology. 2012 Apr;2(1):1-4.
[5] Gould AR, May BK, Elliott WH. Release of extracellular enzymes from Bacillus amyloliquefaciens. Journal of bacteriology. 1975 Apr;122(1):34-40
[6] Hutadilok-Towatana N, Painupong A, Suntinanalert P. Purification and characterization of an extracellular protease from alkaliphilic and thermophilic Bacillus sp. PS719. Journal of bioscience and bioengineering. 1999 Jun;87(5):581-7.doi:10.1016/ S1389-1723 (99)80118-2.
[7] Malik M, Kain J, Pettigrew C, Ogram A. Purification and molecular analysis of microbial DNA from compost. Journal of Microbiological Methods. 1994 Sep;20 (3):183-96.doi:10.1016/ 0167-7012(94)90 003-5.
[8] Xu D, Coˆte´ JC. Phylogenetic relationships between Bacillus species and related genera inferred from comparison of 39 end 16S rDNA and 59 end 16S–23S ITS nucleotide sequences. International Journal of Systematic and Evolutionary Microbiology 2003 Sep;53:695–704.doi: 10.1099/ ijs.0.02346-0.
[9] Wang W, Sun M. Phylogenetic relationships between Bacillus species and related genera inferred from 16s rDNA sequences. Brazilian Journal of Microbiology. 2009 May;40:505-21.38
[10] Vidyasagar M, Prakas S, Jayalakshmi SK, Sreeramulu K. Optimization of culture conditions for the production of halothermophilic protease from h a l o p h i l i c b a c t e r i u m Chromohalobacter sp. TVSP101. World Journal of Microbiology and Biotechnology. 2007 May;23: 655–62.doi:10.1007/s11274- 006-9279-1.
[11] Cheng SW, Wang YF, Wanga ML. Statistical optimization of medium compositions for alkaline protease production by newly isolated. Chemical and Biochemical Engineering 2012 Jul;26(3):225-31. (12) Son ES, Kim JI. Purification and characterization of caseinolytic extracellular protease from Bacillus amyloliquefaciens S94. Journal of Microbiology 2002 Mar;40(1):26-32.
[13] Cho SJ, Park JH, Park SJ, Lim JS, Kim EH, Cho YJ, et al. Purification and Characterization of Extracellular Temperature-Stable Serine Protease from Aeromonas hydrophila. Journal of Microbiology. 2003 Sep;41(3):207-11.
[14] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry. 1951 Nov;193(1): 265-75.
[15] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680-5. doi:0.1038/227680a 0.
[16] Steinberg TH. Protein gel staining methods: an introduction and overview. Methods in enzymology. 2009;463:541-63.doi:10.1016/ S0076-6879(09)63031-7.
[17] Garcia-Carreno FL, Dimes LE, Haard NF. Substrate-gel electrophoresis for composition and molecular weight of proteinases or proteinaceous proteinase inhibitors. Analytical biochemistry. 1993 Oct;214(1):65-9.doi:10.1006/ abio.1993.1 457.
[18] Wilkesman J, Kurz L. Protease analysis by zymography: a review on techniques and patents. Recent patents on biotechnology. 2009;3(3):175-84. doi:10.2174/18722080 9789389162.
[19] Fernanda S, Susana J, Anita E, Vanja K, Artur CP, Gubitz GM. A novel metalloprotease from Bacillus cereus for protein fibre processing. Enzyme Microbial Technology. 2007 Jun;40(7): 177281.doi:10.1016/j. enzmictec.2006.12 .017.
[20] Deng A, Wu J, Zhang Y, Zhang G, Wen T. Purification and characterization of a surfactant- stable high-alkaline protease from Bacillus sp. B001. Bioresource technology. 2010 Sep;101(18): 7111-7.doi: 10.1016/j. biortech.2010.03.130.
[21] Charles P, Devanathan V, Anbu P, Ponnuswamy MN, Kalaichelvan PT, Hur BK. Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10. Journal of basic microbiology. 2008 Oct;48(5):347-52.doi:10.1002/ jobm.2008 00043.
[22] Phrommao E, Yongsawatdigul J, Rodtong S, Yamabhai M. A novel subtilase with NaCl-activated and oxidant-stable activity from Virgibacillus sp. SK37. Biomedical Biotechnology. 2011 Jun;11:65-80. doi: 10.1186/1472-6750-11-65.
[23] Vishwanatha KS, Appu-Rao AG, Singh SA. Production and characterization of a milk-clotting enzyme from Aspergillus oryzae MTCC 5341. Applied microbiology and biotechnology. 2010 Feb;85(6):1849-59.doi:10.1007/ s00253-0 09-2197-z.
[24] Castro GR. Enzymatic activities of proteases dissolved in organic solvents. Enzyme Microbial Technology. 1999 Nov;25:689–94. doi:10.1016/S0141-0229 (99)00099-X.
[25] Saborowski R, Sahling G, Navarette-del-Toro MA, Walter I, Garc´-Carreño FL. Stability and effects of organic solvents on endopeptidases from the gastric fluid of the marine crab Cancer pagurus. Journal of Molecular Catalysis B. 2004 Aug;30:109-18. doi:10.1016/j.molcatb.2004.04.002.
[26] Verma KS, Ghosh KK. Catalytic activity of enzyme in water/organic cosolvent mixture for the hydrolysis of p-nitrophenyl acetate and p-nitrophenyl benzoate. Indian Journal of Chemistry 2010 Aug;49(A): 1041-6.
[27] Shevchenko A, Tomas H, Havlis J, Olsen JV, Mann M. In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nature protocols. 2006 Jan;1(6):2856-5860.doi:10.1038/ nprot.20 06.468.
[28] Shaheen M, Alishah A, Hameed A, Hasan F. Influence of culture conditions on production and activity of protease from Bacillus subtilis BS1. Pakistan Journal of Botany. 2008;40(5):2161-9.
[29] Shivanand P, Jayaraman G. Isolation and characterization of a metal ion-dependent alkaline protease from a halotolerant Bacillus aquimaris VITP4. Indian Journal of Biochemistry and Biophysics 2011 Apr;48(2):95-100.