Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis

© 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 subtili...

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Main Authors: Shinji Takenaka, Jyun Yoshinami, Ampin Kuntiya, Charin Techapun, Noppol Leksawasdi, Phisit Seesuriyachan, Thanongsak Chaiyaso, Masanori Watanabe, Kosei Tanaka, Ken ichi Yoshida
Format: Journal
Published: 2018
Online Access:https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85031898530&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/43577
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-435772018-01-24T03:50:17Z Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis Shinji Takenaka Jyun Yoshinami Ampin Kuntiya Charin Techapun Noppol Leksawasdi Phisit Seesuriyachan Thanongsak Chaiyaso Masanori Watanabe Kosei Tanaka Ken ichi Yoshida © 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. 2018-01-24T03:50:17Z 2018-01-24T03:50:17Z 2017-10-16 Journal 15736776 01415492 2-s2.0-85031898530 10.1007/s10529-017-2459-2 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85031898530&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/43577
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
description © 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.
format Journal
author Shinji Takenaka
Jyun Yoshinami
Ampin Kuntiya
Charin Techapun
Noppol Leksawasdi
Phisit Seesuriyachan
Thanongsak Chaiyaso
Masanori Watanabe
Kosei Tanaka
Ken ichi Yoshida
spellingShingle Shinji Takenaka
Jyun Yoshinami
Ampin Kuntiya
Charin Techapun
Noppol Leksawasdi
Phisit Seesuriyachan
Thanongsak Chaiyaso
Masanori Watanabe
Kosei Tanaka
Ken ichi Yoshida
Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis
author_facet Shinji Takenaka
Jyun Yoshinami
Ampin Kuntiya
Charin Techapun
Noppol Leksawasdi
Phisit Seesuriyachan
Thanongsak Chaiyaso
Masanori Watanabe
Kosei Tanaka
Ken ichi Yoshida
author_sort Shinji Takenaka
title Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis
title_short Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis
title_full Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis
title_fullStr Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis
title_full_unstemmed Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis
title_sort characterization and mutation analysis of a halotolerant serine protease from a new isolate of bacillus subtilis
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85031898530&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/43577
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