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A recombinant subtilisin with keratinolytic and fibrin(ogen)olytic activity

Paper ID Volume ID Publish Year Pages File Format Full-Text
34371 45020 2014 7 PDF Available
Title
A recombinant subtilisin with keratinolytic and fibrin(ogen)olytic activity
Abstract

•A recombinant subtilisin displaying keratinolytic and fibrin(ogen)olytic activity.•A subtilisin E homologous keratinase posses fibrinogen(olytic) activity.•The effect of mutations known as increasing thermal stability in subtilins seems to be neutralized by a His-tag tail.•A His-tag tail at C-terminal improves thermal stability in a subtilisin.

KerS14 is a keratinase with great potential in tannery, since it degrades keratin without damaging collagen, a feature suitable for various industrial uses. The enzyme was previously characterized and described as a serino endopeptidase belonging to the subtilisin group. However, KerS14 low thermal stability impairs its biotechnological potential. The present work presents several attempts to improve KerS14 thermal stability. KerS14 ORF was cloned into pET-5a vector with a His-tag at C-terminal, and four different mutants enzymes (G61C, S98C, P239R and G61C-S98C) were produced by site-direct mutagenesis. The recombinant enzyme and four mutants were expressed, purified and characterized regarding their thermal stability, optimum temperature and pH. The presence of a His-tag was shown to increase the KerS14 thermal stability, and to decrease the thermal stability of mutant enzymes. In addition, the recombinant enzyme has a remarkable fibrin(ogen)olytic activity. This indicates that the enzyme has a potential for application in cardiovascular diseases, besides its use in tanning as a dehairing agent.

Keywords
Bacillus subtilis S14; Keratinase; Fibrin(ogen)olytic activity; His-tag
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A recombinant subtilisin with keratinolytic and fibrin(ogen)olytic activity
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 49, Issue 6, June 2014, Pages 948–954
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
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