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- Title
Improvement in the thermostability of chitosanase from Bacillus ehimensis by introducing artificial disulfide bonds.
- Authors
Sheng, Jun; Ji, Xiaofeng; Zheng, Yuan; Wang, Zhipeng; Sun, Mi
- Abstract
Objective: To determine the effects of artificial disulfide bridges on the thermostability and catalytic efficiency of chitosanase EAG1. Results: Five artificial disulfide bridges were designed based on the structural information derived from the three-dimensional (3-D) model of chitosanase EAG1. Two beneficial mutants (G113C/D116C, A207C-L286C) were located in the flexible surface loop region, whereas the similar substitutions introduced in α-helices regions had a negligible effect. Mut5, the most active mutant, had a longer half-life at 50 °C (from 10.5 to 69.3 min) and a 200 % higher catalytic efficiency ( K / K ) than that of the original EAG1. Conclusions: The contribution of disulfide bridges to enzyme thermostability is mainly dependent on its location within the polypeptide chain. Strategical placement of a disulfide bridge in flexible regions provides a rigid support and creation of a protected microenvironment, which is effective in improving enzyme's thermostability and catalytic efficiency.
- Subjects
BACILLUS (Bacteria); BACTERIAL enzyme analysis; THERMAL stability; BACTERIAL mutation; POLYPEPTIDES
- Publication
Biotechnology Letters, 2016, Vol 38, Issue 10, p1809
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-016-2168-2