title: Constitutive expression of the DUR1,2 gene in an industrial yeast strain to minimize ethyl carbamate production during Chinese rice wine fermentation.
abstract: Urea and ethanol are the main precursors of ethyl carbamate (EC) in Chinese rice wine. During fermentation, urea is generated from arginine by arginase in Saccharomyces cerevisiae, and subsequently cleaved by urea amidolyase or directly transported out of the cell into the fermentation liquor, where it reacts with ethanol to form EC. To reduce the amount of EC in Chinese rice wine, we metabolically engineered two yeast strains, N85(DUR1,2) and N85(DUR1,2)-c, from the wild-type Chinese rice wine yeast strain N85. Both new strains were capable of constitutively expressing DUR1,2 (encodes urea amidolyase) and thus enhancing urea degradation. The use of N85(DUR1,2) and N85(DUR1,2)-c reduced the concentration of EC in Chinese rice wine fermented on a small-scale by 49.1% and 55.3%, respectively, relative to fermentation with the parental strain. All of the engineered strains showed good genetic stability and minimized the production of urea during fermentation, with no exogenous genes introduced during genetic manipulation, and were therefore suitable for commercialization to increase the safety of Chinese rice wine.
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China.
authors.forename: Dianhui
authors.initials: D
authors.lastname: Wu
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Li
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China Industrial Technology Research Institute of Jiangnan University in Suqian, 888 Renmin Road, Suqian 223800, P.R. China jlu@jiangnan.edu.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Lu
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, P. R. China National Engineering Research Center for Chinese Rice Wine, China Shaoxing Rice Wine Group Co., Ltd. Shaoxing 312000, P. R. China.
authors.forename: Guangfa
authors.initials: G
authors.lastname: Xie
mesh_terms: D002681:China; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D060847:Metabolic Engineering; D012275:Oryza / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics; D014508:Urea / Q000378:metabolism; D014520:Urethane / Q000378:metabolism*; D014920:Wine / Q000382:microbiology*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D014520:Urethane; D000431:Ethanol; D014508:Urea
keywords: Chinese rice wine; Saccharomyces cerevisiae; DUR1,2; ethyl carbamate; urea