Abstract
High-quality environmentally-friendly bioplastics can be produced by mixing poly-L-lactate with poly-D-lactate. On an industrial scale, this process simultaneously consumes large amounts of both optically pure lactate stereoisomers. However, because optimal growth conditions of L-lactate producers often differ from those of D-lactate producers, each stereoisomer is produced in a specialised facility, which raises cost and lowers sustainability. To address this challenge, we metabolically engineered Lactobacillus gasseri JCM 1131T, a bioprocess-friendly and genetically malleable strain of homofermentative lactic acid bacterium, to efficiently produce either pure L- or pure D-lactate under the same bioprocess conditions. Transformation of L. gasseri with plasmids carrying additional genes for L- or D-lactate dehydrogenases failed to affect the ratio of produced stereoisomers, but inactivation of the endogenous genes created strains which yielded 0.96 g of either L- or D-lactate per gram of glucose. In this study, the plasmid pHBintE, routinely used for gene disruption in Bacillus megaterium, was used for the first time to inactivate genes in lactobacilli. Strains with inactivated genes for endogenous lactate dehydrogenases efficiently fermented sugars released by enzymatic hydrolysis of alkali pre-treated wheat straw, an abundant lignocellulose-containing raw material, producing 0.37–0.42 g of lactate per gram of solid part of alkali-treated wheat straw. Thus, the constructed strains are primed to serve as producers of both optically pure L-lactate and D-lactate in the next-generation biorefineries.
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Data availability
Reported nucleotide sequence data are available in the Third Party Annotation Section of the DDBJ/ENA/GenBank databases under the accession numbers TPA: BK010903-BK010905. L. gasseri JCM 1131T is available from Japan Collection of Microorganisms (JCM).
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This work was supported by the Croatian Science Foundation (Grant Numbers IP-11-2013-9158-SPECH-LRM and IP-01-2018-9717-SPB-LCF, awarded to Prof Božidar Šantek).
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All authors contributed to the study design. Material preparation, data collection and analysis were performed by BŽ, AT, JLP, MSM, AS, NM and MN. BŠ and IKS initiated and coordinated the study. All authors read and approved the final manuscript.
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Žunar, B., Trontel, A., Svetec Miklenić, M. et al. Metabolically engineered Lactobacillus gasseri JCM 1131 as a novel producer of optically pure L- and D-lactate. World J Microbiol Biotechnol 36, 111 (2020). https://doi.org/10.1007/s11274-020-02887-2
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DOI: https://doi.org/10.1007/s11274-020-02887-2