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Engineering Escherichia coli for a high yield of 1,3-propanediol near the theoretical maximum through chromosomal integration and gene deletion

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Abstract

Glycerol dehydratase (gdrAB-dhaB123) operon from Klebsiella pneumoniae and NADPH-dependent 1,3-propanediol oxidoreductase (yqhD) from Escherichia coli were stably integrated on the chromosomal DNA of E. coli under the control of the native-host ldhA and pflB constitutive promoters, respectively. The developed E. coli NSK015 (∆ldhA::gdrAB-dhaB123 ∆ackA::FRT ∆pflB::yqhD ∆frdABCD::cat-sacB) produced 1,3-propanediol (1,3-PDO) at the level of 36.8 g/L with a yield of 0.99 mol/mol of glycerol consumed when glucose was used as a co-substrate with glycerol. Co-substrate of glycerol and cassava starch was also utilized for 1,3-PDO production with the concentration and yield of 31.9 g/L and 0.84 mol/mol of glycerol respectively. This represents a work for efficient 1,3-PDO production in which the overexpression of heterologous genes on the E. coli host genome devoid of plasmid expression systems. Plasmids, antibiotics, IPTG, and rich nutrients were omitted during 1,3-PDO production. This may allow a further application of E. coli NSK015 for the efficient 1,3-PDO production in an economically industrial scale.

Key points

 • gdrAB-dhaB123 and yqhD were overexpressed in E. coli devoid of a plasmid system

• E. coli NSK015 produced a high yield of 1,3-PDO at 99% theoretical maximum

• Cassava starch was alternatively used as substrate for economical 1,3-PDO production

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Acknowledgements

Author thanks the Thailand Research Fund (TRF) under the Royal Golden Jubilee PhD scholarship (Grant No. PHD/0125/2556) that provided a financial support for this work.

Funding

This study was funded by Thailand Research Fund (TRF) under the Royal Golden Jubilee PhD scholarship (Grant No. PHD/0125/2556).

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Authors and Affiliations

  1. Metabolic Engineering Research Unit, School of Biotechnology, Institute of Agricultural Technology, Suranaree Sub-District, Suranaree University of Technology, 111 University Avenue, Muang district, Nakhon Ratchasima, 30000, Thailand

    Nonthaporn Wong & Kaemwich Jantama

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  1. Nonthaporn Wong
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  2. Kaemwich Jantama
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Contributions

KJ conceived, designed research, and performed project administration and funding acquisition. NW conducted experiments and wrote an original draft of the manuscript. KJ also analyzed data, provided comments, and edited and reviewed the final manuscript. All authors read and approved the manuscript.

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Correspondence to Kaemwich Jantama.

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Wong, N., Jantama, K. Engineering Escherichia coli for a high yield of 1,3-propanediol near the theoretical maximum through chromosomal integration and gene deletion. Appl Microbiol Biotechnol 106, 2937–2951 (2022). https://doi.org/10.1007/s00253-022-11898-y

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  • DOI: https://doi.org/10.1007/s00253-022-11898-y

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