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Development of a Novel D-Lactic Acid Production Platform Based on Lactobacillus saerimneri TBRC 5746

  • Systems and Synthetic Microbiology and Bioinformatics
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Abstract

D-Lactic acid is a chiral, three-carbon organic acid, that bolsters the thermostability of polylactic acid. In this study, we developed a microbial production platform for the high-titer production of D-lactic acid. We screened 600 isolates of lactic acid bacteria (LAB) and identified twelve strains that exclusively produced D-lactic acid in high titers. Of these strains, Lactobacillus saerimneri TBRC 5746 was selected for further development because of its homofermentative metabolism. We investigated the effects of high temperature and the use of cheap, renewable carbon sources on lactic acid production and observed a titer of 99.4 g/L and a yield of 0.90 g/g glucose (90% of the theoretical yield). However, we also observed L-lactic acid production, which reduced the product’s optical purity. We then used CRISPR/dCas9-assisted transcriptional repression to repress the two Lldh genes in the genome of L. saerimneri TBRC 5746, resulting in a 38% increase in D-lactic acid production and an improvement in optical purity. This is the first demonstration of CRISPR/dCas9-assisted transcriptional repression in this microbial host and represents progress toward efficient microbial production of D-lactic acid.

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Acknowledgements

The project was funded by the Integrated Technology Platform (Biobased Materials) project, “IBMDL1-High-level Microbial Production of Enantiomerically Pure D-lactic Acid,” supported by the National Science and Technology Development Agency [grant number P-17-522777].

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Author notes

  1. Pipat Sudying

    Present address: Chulabhorn Research Institute, Laksi, Bangkok, 10210, Thailand

Authors and Affiliations

  1. National Center for Genetic Engineering and Biotechnology, Klong Luang, Pathum Thani, 12120, Thailand

    Kitisak Sansatchanon, Pipat Sudying, Peerada Promdonkoy, Yutthana Kingcha, Wonnop Visessanguan, Sutipa Tanapongpipat, Weerawat Runguphan & Kanokarn Kocharin

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  1. Kitisak Sansatchanon
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Contributions

Conceptualization, WR, WV and KK; methodology, WR and KK; validation, KS, PS, YK, KK and WR; formal analysis, KK and WR; investigation, KS, PS and PP; resources, WR; data curation, KS, PS, YK, KK and WR; writing—original draft preparation, WR and KK; writing—review and editing, WR, KK and PP; visualization, WR and KK; supervision, WV, ST, WR and KK; project administration, WR and KK; funding acquisition, WR All authors have read and agreed to the published version of the manuscript.”

Corresponding author

Correspondence to Kanokarn Kocharin.

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The authors declare no conflict of interest. The funder had no role in the design of the study; in the collection, analysis, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

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Sansatchanon, K., Sudying, P., Promdonkoy, P. et al. Development of a Novel D-Lactic Acid Production Platform Based on Lactobacillus saerimneri TBRC 5746. J Microbiol. 61, 853–863 (2023). https://doi.org/10.1007/s12275-023-00077-x

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  • DOI: https://doi.org/10.1007/s12275-023-00077-x

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