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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.

Efficient production of acetoin in Saccharomyces cerevisiae by disruption of 2,3-butanediol dehydrogenase and expression of NADH oxidase