Stability enhancement of urethanase from Lysinibacillus fusiformis by site-directed mutagenesis.

Xiaohui Liu; Fang Fang; Xiaole Xia; Guocheng DU; Jian Chen

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Stability enhancement of urethanase from Lysinibacillus fusiformis by site-directed mutagenesis.

Author: Xiaohui LIU ¹ ; Fang FANG ¹ ; Xiaole XIA ¹ ; Guocheng DU ² ; Jian CHEN ¹
Author Information

1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
2. Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi 214122, Jiangsu, China.
Publication Type:Journal Article
Keywords: enzyme stability; ethyl carbamate; site-directed mutagenesis; urethanase
MeSH: Amidohydrolases; biosynthesis; genetics; Bacillaceae; enzymology; genetics; Bacterial Proteins; biosynthesis; genetics; Computer-Aided Design; Enzyme Stability; Ethanol; Mutagenesis, Site-Directed; Protein Engineering
From: Chinese Journal of Biotechnology 2016;32(9):1233-1242
CountryChina
Language:Chinese
Abstract: Ethyl carbamate as a potential carcinogen commonly exists in traditional fermented foods and beverages. Enzymatic removal of ethyl carbamate from fermented foods and beverages is an efficient and safe method. In this study, we mutated urethanase from Lysinibacillus fusiformis SC02 on the Q328 site through computer aided design approaches. The half-life of resulting mutants Q328C and Q328V was detected to be 7.46 and 1.96 folds higher than that of the original enzyme, and Q328R presented better thermal-tolerance than the original urethanase when incubated at high temperature. The tolerance of Q328C to ethanol and acid also increased when compared with that of the original enzyme. The stability and tolerance to acid and ethanol of urethanase could be improved by modification on its Q328 site.