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Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11
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Type
Academic journal
Author
Journal
한국축산학회(구 한국동물자원과학회) 한국축산학회지 한국축산학회지 제51권 제6호 KCI Excellent Accredited Journal
Published
2009.1
Pages
527 - 536 (10page)

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Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11
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A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production(1.64 units/ml) was attained in a shake flask culture when the isolate was grown at 40℃, for 32 h in basal medium supplemented with starch(0.25%) and gelatin(1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and 50℃, respectively. Ca2+ and Mn2+ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below 50℃, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity(6.74 units/mg) than the parent strain IB No. 11 when grown at 40℃ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.
#Protease #Bacillus subtilis IB No. 11 #Isolation #Characterization

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References (23)

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Anisa / 2009 / A novel surfactant-stable alkaline serine- protease from a newly isolated Bacillus mojavensis / Process Biochemistry : 4429 ~ 4435 google schola Bascaran, V / 1990 / Regulation of extracellular protease production in Streptomyces clavuligerus / Appl. Microbiol. Biotechnol 34 : 208 ~ 213 google schola Bradford, M. M / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 72 google schola Chandra, M / 2009 / Development of a mutant of Trichoderma citrinoviride for enhanced production cellualases / Biores. Technol 100 : 1659 ~ 1662 google schola Cupp-Enyard, C / 2008 / Sigma’s non-specific protease activity assay-Casein as a substrate / J. of visualized experiments JoVE 19 : 1 ~ 2 google schola

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