Engineering a newly isolated Bacillus licheniformis strain for the production of (2R,3R)-butanediol

Several microorganisms can produce 2,3-butanediol (BDO), an industrially promising chemical. In this study, a Bacillus licheniformis named as 4071, was isolated from soil sample. It is a GRAS (generally recognized as safe) strain and could over-produce 2,3-BDO. Due to its mucoid forming characteristics, UV-random mutagenesis was carried out to obtain a mucoid-free strain, 4071-15. As a result, capabilities of 4071-15 strain in terms of transformation efficiency of bacillus plasmids (pC194, pUB110, and pUCB129) and fermentation performance were highly upgraded compared to those of the parent strain. In particular, 4071-15 strain could produce 123 g/L of 2,3-BDO in a fed-batch fermentation in which the ratio of (2R,3S)- to (2R,3R)-form isomers was 1:1. To increase the selectivity of (2R,3R)-BDO, budC gene was deleted by using temperature-sensitive gene deletion process via homologous recombination. The 4071-15 △budC mutant strain dramatically increased selectivity of (2R,3R)-BDO to 91% [96.3 g/L of (2R,3R)-BDO and 9.33 g/L of (2R,3S)-BDO], which was 43% higher than that obtained by the parent strain. This study has shown the potential of an isolate for 2,3-BDO production, and that the ratio of 2,3-BDO can be controlled by genetic engineering depending on its industrial usage.