Serratia marcescens has been proved to be a potential strain for industrial 2,3-butanediol production for its high yield, productivity, and other advantages. In this study, the genes slaA, slaB, slaC, and slaR were successfully cloned which were further confirmed to be encoding acetolactate decarboxylase, acetolactate synthase, 2,3-butanediol dehydrogenase, and a LysR-like regulator, respectively. Unlike in Klebsiella sp. or Klebsiella pneumonie and Vibrio sp. or Vibrio cholerae, the gene slaC is separated from other genes. Then it showed that two regulators, SwrR and SlaR, are in charge of this process by exerting effect on the transcription of genes slaA and slaB. By contrast, the expression of gene slaC is unaffected by the two regulators. It means that these two regulators affect the production of 2,3-butanediol by regulating the production of acetoin. Based on these findings, we successfully accelerated the 2,3-butanediol production by inactivation of gene swrR. The obtained results and further investigations should lead to a more suitable fermentation strategy and strain improvement which would be applicable to the industrial production of 2,3-butanediol.