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J Agric Food Chem. 2018 Aug 29;66(34):9061-9069. doi: 10.1021/acs.jafc.8b01313. Epub 2018 Jun 26.

Adaptive Evolution Relieves Nitrogen Catabolite Repression and Decreases Urea Accumulation in Cultures of the Chinese Rice Wine Yeast Strain Saccharomyces cerevisiae XZ-11.

Zhang W1,2,3, Cheng Y1, Li Y4, Du G1, Xie G5, Zou H6, Zhou J1,2,3, Chen J1,3.

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Key Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology , Jiangnan University , 1800 Lihu Road , Wuxi , Jiangsu 214122 , China.
National Engineering Laboratory for Cereal Fermentation Technology , Jiangnan University , 1800 Lihu Road , Wuxi , Jiangsu 214122 , China.
Jiangsu Provisional Research Center for Bioactive Product Processing Technology , Jiangnan University , 1800 Lihu Road , Wuxi , Jiangsu 214122 , China.
Department of Bioengineering, School of Food Sciences and Biotechnology , Zhejiang Gongshang University , Hangzhou 310018 , China.
College of Shaoxing Rice Wine , Zhejiang Shuren University , Shaoxing 312028 , China.
Zhejiang Guyuelongshan Shaoxing Wine Company , 13 Yangjiang Road , Shaoxing , Zhejiang China.


Urea is the major precursor of ethyl carbamate in Chinese rice wine. Although efforts have been made to decrease urea accumulation, few methods can be applied to industrial food production due to potential safety concerns. In this study, adaptive laboratory evolution (ALE) followed by high-throughput screening was used to identify low urea-accumulating strains derived from the industrial Chinese rice wine yeast strain Saccharomyces cerevisiae XZ-11. Three evolved strains were obtained that had 47.9%, 16.6%, and 12.4% lower urea concentrations than the wild-type strain. Comparative genomics analysis revealed that genes involved in carbon and nitrogen metabolism evolved quickly. Transcription levels of genes involved in urea metabolism were dramatically upregulated after ALE. This work describes a novel and safe strategy to improve nitrogen utilization of industrial yeast strains involved in food fermentation. The identified genomic variations may also help direct rational genetic engineering of nitrogen metabolism processes to achieve other goals.


RT-qPCR; Saccharomyces cerevisiae; adaptive laboratory evolution; comparative genomics; high-throughput screening; urea

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