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|Title:||Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis|
Ken ichi Yoshida
|Keywords:||Biochemistry, Genetics and Molecular Biology|
|Abstract:||© 2017, Springer Science+Business Media B.V. Objectives: A bacterial halotolerant enzyme was characterized to understand the molecular mechanism of salt adaptation and to explore its protein engineering potential. Results: Halotolerant serine protease (Apr_No16) from a newly isolated Bacillus subtilis strain no. 16 was characterized. Multiple alignments with previously reported non-halotolerant proteases, including subtilisin Carlsberg, indicated that Apr_No16 has eight acidic or polar amino acid residues that are replaced by nonpolar amino acids in non-halotolerant proteases. Those residues were hypothesized to be one of the primary contributors to salt adaptation. An eightfold mutant substituted with Ala residues exhibited 1.2- and 1.8-fold greater halotolerance at 12.5% (w/v) NaCl than Apr_No16 and Carlsberg, respectively. Amino acid substitution notably shifted the theoretical pI of the eightfold mutant, from 6.33 to 9.23, compared with Apr_No16. The resulting protein better tolerated high salt conditions. Conclusions: Changing the pI of a bacterial serine protease may be an effective strategy to improve the enzyme’s halotolerance.|
|Appears in Collections:||CMUL: Journal Articles|
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