The combination of glycerol metabolic engineering and drug resistance marker-aided genome shuffling to improve very-high-gravity fermentation performances of industrial Saccharomyces cerevisiae
The combination of glycerol metabolic engineering and drug resistance marker-aided genome shuffling to improve very-high-gravity fermentation performances of industrial Saccharomyces cerevisiae
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DOI：
10.1016/j.biortech.2011.12.147
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发表时间：
2012-03-01
影响因子：
11.4
通讯作者：
Wu, Xue-Chang
中科院分区：
文献类型：
--
作者：
Wang, Pin-Mei;Zheng, Dao-Qiong;Wu, Xue-Chang
通过代谢工程和基因组改组的组合克服了与在极高重力（VHG）条件下乙醇生产率相关的挑战，优化了工业酵母菌株的多性状（即副产物形成和胁迫耐受性）。首先，将甘油输出蛋白Fps 1 p缺失到工业菌株Y12中，并异源表达甘油醛-3-磷酸脱氢酶，得到甘油产量较低的修饰菌株YFG 12。其次，对YFG 12进行了三轮耐药标记辅助基因组改组，以提高其乙醇耐受性，并获得了最佳改组菌株TS 5。与野生菌株Y12相比，重组菌株TS 5不仅甘油生成量降低了14.8%，而且发酵速率和乙醇产量分别提高了3.7%和7.6%。此外，本研究提出的基因改造和Cre/loxP辅助3种不同耐药性标记的体系，显著提高了原养型工业微生物的育种效率，为原养型工业微生物育种技术的应用提供了便利。(C)2012爱思唯尔有限公司保留所有权利。
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. (C) 2012 Elsevier Ltd. All rights reserved.