The combination of glycerol metabolic engineering and drug resistance marker-aided genome shuffling to improve very-high-gravity fermentation performances of industrial Saccharomyces cerevisiae

Bioresource Technology
Pin-Mei WangXue-Chang Wu

Abstract

A challenge associated with the ethanol productivity under very-high-gravity (VHG) conditions, optimizing multi-traits (i.e. byproduct formation and stress tolerance) of industrial yeast strains, is overcome by a combination of metabolic engineering and genome shuffling. First, industrial strain Y12 was deleted with a glycerol exporter Fps1p and hetero-expressed with glyceraldehydes-3-phosphate dehydrogenase, resulting in the modified strain YFG12 with lower glycerol yield. Second, YFG12 was subjected to three rounds of drug resistance marker-aided genome shuffling to increase its ethanol tolerance, and the best shuffled strain TS5 was obtained. Compared with wild strain Y12, shuffled strain TS5 not only decreased glycerol formation by 14.8%, but also increased fermentation rate and ethanol yield by 3.7% and 7.6%, respectively. Moreover, the system of genetic modification and Cre/loxP in aid of three different drug-resistance markers presented in the study significantly improved breeding efficiency and will facilitate the application of breeding technologies in prototrophic industrial microorganisms.

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Citations

May 10, 2013·Journal of Industrial Microbiology & Biotechnology·Ji ZhangWensheng Xiang
Dec 19, 2013·Applied Microbiology and Biotechnology·Dao-Qiong ZhengXue-Chang Wu
Mar 7, 2014·Applied Microbiology and Biotechnology·Damien Biot-Pelletier, Vincent J J Martin
Apr 25, 2013·Current Opinion in Biotechnology·Jens NielsenJack Pronk
Apr 15, 2014·FEMS Microbiology Reviews·Jan SteenselsKevin J Verstrepen
Jan 13, 2018·Frontiers in Bioengineering and Biotechnology·Kentaro InokumaAkihiko Kondo
May 22, 2017·Applied Microbiology and Biotechnology·Jens NielsenIvan Mijakovic
Oct 24, 2019·World Journal of Microbiology & Biotechnology·Ping YuTingting Yan

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