D-2,3-butanediol has many industrial applications such as chiral reagents, solvents, anti-freeze agents, and low freezing point fuels. Traditional D-2,3-butanediol producing microorganisms, such as Klebsiella pneumonia and K. xoytoca, are pathogenic and not capable of producing D-2,3-butanediol at high optical purity. Bacillus licheniformis is a potential 2,3-butanediol producer but the wild type strain (WX-02) produces a mix of D- and meso-type isomers. BudC in B. licheniformis is annotated as 2,3-butanediol dehydrogenase or acetoin reductase, but no pervious experiment was performed to verify this hypothesis.

2,3-Butanediol (2,3-BD) can be used as a liquid fuel additive to replace petroleum oil, and as an important platform chemical in the pharmaceutical and plastic industries. Microbial production of 2,3-BD by Bacillus licheniformis presents potential advantages due to its GRAS status, but previous attempts to use this microorganism as a chassis strain resulted in the production of a mix of D-2,3-BD and meso-2,3-BD isomers. The aim of this work was to develop an engineered strain of B. licheniformis suited to produce the high titers of the pure meso-2,3-BD isomer. Glycerol dehydrogenase (Gdh) was identified as the catalyst for D-2,3-BD biosynthesis from its precursor acetoin in B. licheniformis. The gdh gene was, therefore, deleted from the wild-type strain WX-02 to inhibit the flux of acetoin to D-2,3-BD biosynthesis. The acoR gene involved in acetoin degradation through AoDH ES was also deleted to provide adequate flux from acetoin towards meso-2,3-BD. By re-directing the carbon flux distribution, the double-deletion mutant WX-02ΔgdhΔacoR produced 28.2 g/L of meso-2,3-BD isomer with >99 % purity. The titer was 50 % higher than that of the wide type. A bench-scale fermentation by the double-deletion mutant was developed to further improve meso-2,3-BD production. In a fed-batch fermentation, meso-2,3-BD titer reached 98.0 g/L with a purity of >99.0 % and a productivity of 0.94 g/L–h. This work demonstrates the potential of producing meso-2,3-BD with high titer and purity through metabolic engineering of B. licheniformis.

Litchi pomace, a by-product of litchi processing, is rich in dietary fiber. Soluble and insoluble dietary fibers were extracted from litchi pomace, and insoluble dietary fiber was modified by ultrasonic enzymatic treatment to obtain modified soluble and insoluble dietary fibers. The structural, physicochemical, and functional properties of the dietary fiber samples were evaluated and compared. It was found that all dietary fiber samples displayed typical polysaccharide absorption spectra, with arabinose being the most abundant monosaccharide component. Soluble dietary fibers from litchi pomace were morphologically fragmented and relatively smooth, with relatively high swelling capacity, whereas the insoluble dietary fibers possessed wrinkles and porous structures on the surface, as well as higher water holding capacity. Additionally, soluble dietary fiber content of litchi pomace was successfully increased by 6.32 ± 0.14% after ultrasonic enzymatic modification, and its arabinose content and apparent viscosity were also significantly increased. Further, the soluble dietary fibers exhibited superior radical scavenging ability and significantly stimulated the growth of probiotic bacterial species. Taken together, this study suggested that dietary fiber from litchi pomace could be a promising ingredient for functional foods industry.