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Production and characterization of NaCl-activated proteinases from Virgibacillus sp. SK33 isolated from fish sauce fermentation

Paper ID Volume ID Publish Year Pages File Format Full-Text
35591 45097 2008 8 PDF Available
Title
Production and characterization of NaCl-activated proteinases from Virgibacillus sp. SK33 isolated from fish sauce fermentation
Abstract

Virgibacillus sp. SK33 newly isolated from one-month-old Thai fish sauce was studied for its proteinase production. Neopeptone broth (NEO), halobacterium broth (HL) and HL without either yeast extract (HL-Y), peptone (HL-P) or casamino acid (HL-C) were found to be suitable for proteinase production, whereas fish broth (FB) and skim milk salts broth (SKS) appeared to suppress proteinase production. Moreover, yeast extract, peptone and casamino acid equally stimulated proteinase production. The optimal enzyme production for Virgibacillus sp. SK33 was at 5% NaCl, 40 °C. Maximum proteinase production was achieved at 36 h and maximum cell growth was obtained at 72 h in the modified HL supplemented with only yeast extract (Ym). Extracellular proteinase of Virgibacillus sp. SK33 exhibited optimum activity at 50 °C and pH 8, 10 and 11. Crude proteinase was strongly inhibited by phenylmethanesulfonyl fluoride (PMSF), indicating the presence of serine alkaline proteinase. The enzymes preferentially cleaved Suc-Ala-Ala-Pro-Phe-AMC, a substrate for subtilisin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) activity staining showed molecular weights of 56, 46, 42, 32, 25 and 19 kDa. All proteinases exhibited caseinolytic activity at 25% NaCl. Proteolytic activity towards synthetic substrate increased about 4 times in the presence of 25% NaCl, indicating the characteristic of NaCl-activated proteinase. In addition, crude proteinase from Virgibacillus sp. SK33 showed higher proteolytic activity towards anchovy than did commercial proteinases at 25% NaCl, demonstrating its potential for protein hydrolysis at high salt content.

Keywords
Virgibacillus sp.; Fish sauce; Serine proteinase; NaCl-activated proteinase
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Production and characterization of NaCl-activated proteinases from Virgibacillus sp. SK33 isolated from fish sauce fermentation
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 43, Issue 2, February 2008, Pages 185–192
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us