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

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.