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Mutant Potential Ubiquitination Sites in Dur3p Enhance the Urea and Ethyl Carbamate Reduction in a Model Rice Wine System

Zhang Peng, Du Guocheng, Zou Huijun, Xie Guangfa, Chen Jian, Shi Zhongping, Zhou Jingwen
Journal of agricultural and food chemistry 2017 v.65 no.8 pp. 1641-1648
Saccharomyces cerevisiae, arginine, carcinogens, endocytosis, engineering, ethyl carbamate, glutamine, lysine, models, mutants, mutation, nitrogen, plasma membrane, rice wines, transporters, ubiquitination, urea, yeasts
Ubiquitination can significantly affect the endocytosis and degradation of plasma membrane proteins. Here, the ubiquitination of a Saccharomyces cerevisiae urea plasma membrane transporter (Dur3p) was altered. Two potential ubiquitination sites, lysine residues K556 and K571, of Dur3p were predicted and replaced by arginine, and the effects of these mutations on urea utilization and formation under different nitrogen conditions were investigated. Compared with Dur3p, the Dur3pK₅₅₆R mutant showed a 20.1% decrease in ubiquitination level in yeast nitrogen base medium containing urea and glutamine. It also exhibited a >75.8% decrease in urea formation in yeast extract–peptone–dextrose medium and 41.3 and 55.4% decreases in urea and ethyl carbamate formation (a known carcinogen), respectively, in a model rice wine system. The results presented here show that the mutation of Dur3p ubiquitination sites could significantly affect urea utilization and formation. Modifying the ubiquitination of specific transporters might have promising applications in rationally engineering S. cerevisiae strains to efficiently use specific nitrogen sources.