Objectives: 2,3-Butanediol (2,3-BD) is widely used in several chemical syntheses as well as the manufacture of plastics, solvents, and antifreeze formulations, and can be manufactured by microbial glucose fermentation. Conventional (2,3-BD) fermentation typically has low productivity, yield, and purity, and is expensive for commercial applications. We aimed to delete the lactate dehydrogenase and acetate kinase (ldhA and ack) genes in Klebsiella pneumoniae HD79 by using λRed homologous recombination technology, to eliminate by-products and thereby improve (2,3-BD) production. We also analyzed the resulting gene changes by using transcriptomics.
Results: The yield of (2,3-BD) from the mutant Klebsiella strain was 46.21 g/L, the conversion rate was 0.47 g/g, and the productivity was 0.64 g/L·h, which represented increases of 54.9%, 20.5%, and 106.5% respectively, compared to (WT) strains. Lactate and acetate decreased by 48.2% and 62.8%, respectively. Transcriptomics analysis showed that 4628 genes were differentially expressed (404 significantly up-regulated and 162 significantly down-regulated). Moreover, the (2,3-BD) operon genes were differentially expressed.
Conclusion: Our data showed that the biosynthesis of (2,3-BD) was regulated by inducers (lactate and acetate), a regulator (BudR), and carbon flux. Elimination of acidic by-products by ldhA and ack knockdown significantly improved (2,3-BD) production. Our results provide a deeper understanding of the mechanisms underlying (2,3-BD) production, and form a molecular basis for the improvement this process by genetic modification in the future.
Keywords: 2,3-Butanediol; Acetate kinase; Klebsiella pneumoniae; Lactate dehydrogenase; Transcriptomics.