LAUSR

Abstract Acetoin is a natural flavor with potential applications in food, pharmaceutical, and chemical industries. However, the titer of acetoin in fermentation production of this compound is still relatively low, which hinders its large-scale industrial production. In this work, fermentative production of acetoin by Bacillus licheniformis WX-02 was optimized through metabolic engineering and fermentation control to increase the titer of this compound. First, three single-gene mutants, deficient in genes of 2,3-butanediol dehydrogenase ( budC ), glycerol dehydrogenase ( gdh ), and acetoin dehydrogenase transcriptional activator ( acoR ) respectively, were compared. It was found that single deletion of budC or acoR significantly enhanced the acetoin production. Second, double genes mutant (WX-02Δ budC Δ acoR ) and three genes mutant (WX-02Δ budC Δ gdh Δ acoR ) were evaluated, and the maximum acetoin titer reached 43.95 g L −1 in WX-02Δ budC Δ acoR . Finally, fermentation conditions were investigated for the mutant WX-02Δ budC Δ acoR , and moderate acidic and agitation conditions favored the acetoin production. Using three-stage agitation in fed-batch fermentation, 2,3-butanediol was accumulated to 110.04 g L −1 , and then converted to acetoin. The maximum acetoin titer reached 78.79 g L −1 , much higher than those of reported previously for the B. licheniformis strains. Collectively, this study reported an efficient approach producing acetoin by B. licheniformis strain through metabolic engineering and fermentation optimization.