Efficient production of acetoin in Saccharomyces cerevisiae by disruption of 2,3-butanediol dehydrogenase and expression of NADH oxidase.
Efficient production of acetoin in Saccharomyces cerevisiae by disruption of 2,3-butanediol dehydrogenase and expression of NADH oxidase.
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DOI：
10.1038/srep27667
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发表时间：
2016-06-09
影响因子：
4.6
通讯作者：
Hahn JS
中科院分区：
文献类型：
--
作者：
Bae SJ;Kim S;Hahn JS
乙偶姻作为增味增香剂广泛应用于食品和化妆品工业。对于本研究中的乙偶姻生产，使用酿酒酵母JHY 605作为宿主菌株，其中通过缺失5个醇脱氢酶基因（ADH 1、ADH 2、ADH 3、ADH 4和ADH 5）和2个甘油3-磷酸脱氢酶基因（GPD 1和GPD 2）来大部分消除乙醇和甘油的生产。为了提高乙偶姻的产量，过量表达来自枯草芽孢杆菌的编码α-乙酰乳酸合成酶（AlsS）和α-乙酰乳酸脱羧酶（AlsD）的乙偶姻生物合成基因，并缺失编码将乙偶姻转化为2，3-丁二醇的丁二醇脱氢酶的BDH 1。此外，通过过表达乳酸乳球菌的水形成型NADH氧化酶（NoxE）的NAD+再生，成功地缓解了从葡萄糖生产乙偶姻期间产生的辅因子不平衡。结果表明，工程菌JHY 617-SDN在补料分批发酵中产生100.1g/L乙偶姻，产量为0.44g/g葡萄糖。
Acetoin is widely used in food and cosmetic industry as taste and fragrance enhancer. For acetoin production in this study, Saccharomyces cerevisiae JHY605 was used as a host strain, where the production of ethanol and glycerol was largely eliminated by deleting five alcohol dehydrogenase genes (ADH1, ADH2, ADH3, ADH4, and ADH5) and two glycerol 3-phosphate dehydrogenase genes (GPD1 and GPD2). To improve acetoin production, acetoin biosynthetic genes from Bacillus subtilis encoding α-acetolactate synthase (AlsS) and α-acetolactate decarboxylase (AlsD) were overexpressed, and BDH1 encoding butanediol dehydrogenase, which converts acetoin to 2,3-butanediol, was deleted. Furthermore, by NAD+ regeneration through overexpression of water-forming NADH oxidase (NoxE) from Lactococcus lactis, the cofactor imbalance generated during the acetoin production from glucose was successfully relieved. As a result, in fed-batch fermentation, the engineered strain JHY617-SDN produced 100.1 g/L acetoin with a yield of 0.44 g/g glucose.