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Increased activity of β-glucuronidase variants produced by site-directed mutagenesis.

Authors :
Zhang, Xiaolei
Sitasuwan, Pongkwan
Horvath, Gary
Yang, Jia
Nie, Yuzhe
Marinova, Margarita
Lee, L. Andrew
Wang, Qian
Source :
Enzyme & Microbial Technology. Feb2018, Vol. 109, p20-24. 5p.
Publication Year :
2018

Abstract

β -glucuronidase (BGus) is an essential glycosyl hydrolase which has been widely used in biological and biomedical applications. In this paper, we report the construction and screening of nineteen Escherichia coli BGus (EBGus) mutants using site-directed mutagenesis. The mutants G559N, G559S and G559T showed a 3–5 fold increase in enzyme activity in comparison to wild type EBGus. In particular, G559S, with the highest activity, showed 2–6 fold enhanced activity compared to abalone and snail BGus extracts. Moreover, the glycine to serine mutagenesis for the same site in Staphylococcus sp . RLH1 BGus (StBGus) exhibited significantly enhanced activity, which indicated the importance of the G559 → S mutation on BGus function. Based on this structural analysis, we postulate that the mutation at G559 plays an important role in the stabilization of the enzyme conformation, and thereby facilitates the effective binding of substrate. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
01410229
Volume :
109
Database :
Academic Search Index
Journal :
Enzyme & Microbial Technology
Publication Type :
Academic Journal
Accession number :
126635147
Full Text :
https://doi.org/10.1016/j.enzmictec.2017.09.005