a novel strategy to construct yeast saccharomyces cerevisiae strains for very high gravity fermentation一项新策略来构建酵母发酵酿酒酵母菌株非常高的重力.pdfVIP

A Novel Strategy to Construct Yeast Saccharomyces cerevisiae Strains for Very High Gravity Fermentation Xianglin Tao., Daoqiong Zheng., Tianzhe Liu, Pinmei Wang, Wenpeng Zhao, Muyuan Zhu, Xinhang Jiang, Yuhua Zhao, Xuechang Wu* Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang Province, People’s Republic of China Abstract Very high gravity (VHG) fermentation is aimed to considerably increase both the fermentation rate and the ethanol concentration, thereby reducing capital costs and the risk of bacterial contamination. This process results in critical issues, such as adverse stress factors (ie., osmotic pressure and ethanol inhibition) and high concentrations of metabolic byproducts which are difficult to overcome by a single breeding method. In the present paper, a novel strategy that combines metabolic engineering and genome shuffling to circumvent these limitations and improve the bioethanol production performance of Saccharomyces cerevisiae strains under VHG conditions was developed. First, in strain Z5, which performed better than other widely used industrial strains, the gene GPD2 encoding glycerol 3-phosphate dehydrogenase was deleted, resulting in a mutant (Z5DGPD2) with a lower glycerol yield and poor ethanol productivity. Second, strain Z5DGPD2 was subjected to three rounds of genome shuffling to improve its VHG fermentation performance, and the best performing strain SZ3-1 was obtained. Results showed that strain SZ3-1 not only produced less glycerol, but also increased the ethanol yield by up to 8% compared with the parent strain Z5. Further analysis suggested that the improved ethanol yield in strain SZ3-1 was mainly contributed by the enhanced ethanol tolerance of the strain. The differences in ethanol