Acetoin is an important physiological metabolite excreted by many microorganisms. The excretion of acetoin, which can be diagnosed by the Voges Proskauer test and serves as a microbial classification marker, has its vital physiological meanings to these microbes mainly including avoiding acification, participating in the regulation of NAD/NADH ratio, and storaging carbon. The well-known anabolism of acetoin involves alpha-acetolactat synthase and alpha-acetolactate decarboxylase; yet its catabolism still contains some differing views, although much attention has been focused on it and great advances have been achieved. Current findings in catabolite control protein A (CcpA) mediated carbon catabolite repression may provide a fuller understanding of the control mechanism in bacteria. In this review, we first examine the acetoin synthesis pathways and its physiological meanings and relevancies; then we discuss the relationship between the two conflicting acetoin cleavage pathways, the enzymes of the acetoin dehydrogenase enzyme system, major genes involved in acetoin degradation, and the CcpA mediated acetoin catabolite repression pathway; in the end we discuss the genetic engineering progresses concerning applications. To date, this is the first integrated review on acetoin metabolism in bacteria, especially with regard to catabolic aspects. The apperception of the generation and dissimilation of acetoin in bacteria will help provide a better understanding of microbial strategies in the struggle for resources, which will consequently better serve the utilization of these microbes.

No abstract

The nutritional requirements for acetoin production by Bacillus subtilis CICC 10025 were optimized statistically in shake flask experiments using indigenous agroindustrial by-products. The medium components considered for initial screening in a Plackett-Burman design comprised a-molasses (molasses submitted to acidification pretreatment), soybean meal hydrolysate (SMH), KH(2)PO(4).3H(2)O, sodium acetate, MgSO(4).7H(2)O, FeCl(2), and MnCl(2), in which the first two were identified as significantly (at the 99% significant level) influencing acetoin production. Response surface methodology was applied to determine the mutual interactions between these two components and optimal levels for acetoin production. In flask fermentations, 37.9 g l(-1) acetoin was repeatedly achieved using the optimized concentrations of a-molasses and SMH [22.0% (v/v) and 27.8% (v/v), respectively]. a-Molasses and SMH were demonstrated to be more productive than pure sucrose and yeast extract plus peptone, respectively, in acetoin fermentation. In a 5-l fermenter, 35.4 g l(-1) of acetoin could be obtained after 56.4 h of cultivation. To our knowledge, these results, i.e., acetoin yields in flask or fermenter fermentations, were new records on acetoin fermentation by B. subtilis.

BackgroundThe high costs of pyridine nucleotide cofactors have limited the applications of NAD(P)-dependent oxidoreductases on an industrial scale. Although NAD(P)H regeneration systems have been widely studied, NAD(P)+ regeneration, which is required in reactions where the oxidized form of the cofactor is used, has been less well explored, particularly in whole-cell biocatalytic processes.Methodology/Principal FindingsSimultaneous overexpression of an NAD+ dependent enzyme and an NAD+ regenerating enzyme (H2O producing NADH oxidase from Lactobacillus brevis) in a whole-cell biocatalyst was studied for application in the NAD+-dependent oxidation system. The whole-cell biocatalyst with (2R,3R)-2,3-butanediol dehydrogenase as the catalyzing enzyme was used to produce (3R)-acetoin, (3S)-acetoin and (2S,3S)-2,3-butanediol.Conclusions/SignificanceA recombinant strain, in which an NAD+ regeneration enzyme was coexpressed, displayed significantly higher biocatalytic efficiency in terms of the production of chiral acetoin and (2S,3S)-2,3-butanediol. The application of this coexpression system to the production of other chiral chemicals could be extended by using different NAD(P)-dependent dehydrogenases that require NAD(P)+ for catalysis.

No abstract