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Genome tailoring powered production of isobutanol in continuous CO2/H2 blend fermentation using engineered acetogen biocatalyst

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The cell energy fraction that powered maintenance and expression of genes encoding pro-phage elements, pta-ack cluster, early sporulation, sugar ABC transporter periplasmic proteins, 6-phosphofructokinase, pyruvate kinase, and fructose-1,6-disphosphatase in acetogen Clostridium sp. MT871 was re-directed to power synthetic operon encoding isobutanol biosynthesis at the expense of these genes achieved via their elimination. Genome tailoring decreased cell duplication time by 7.0 ± 0.1 min (p < 0.05) compared to the parental strain, with intact genome and cell duplication time of 68 ± 1 min (p < 0.05). Clostridium sp. MT871 with tailored genome was UVC-mutated to withstand 6.1 % isobutanol in fermentation broth to prevent product inhibition in an engineered commercial biocatalyst producing 5 % (674.5 mM) isobutanol during two-step continuous fermentation of CO2/H2 gas blend. Biocatalyst Clostridium sp. MT871RG11IBR6 was engineered to express six copies of synthetic operon comprising optimized synthetic format dehydrogenase, pyruvate formate lyase, acetolactate synthase, acetohydroxyacid reductoisomerase, 2,3-dihydroxy-isovalerate dehydratase, branched-chain alpha-ketoacid decarboxylase gene, aldehyde dehydrogenase, and alcohol dehydrogenase, regaining cell duplication time of 68 ± 1 min (p < 0.05) for the parental strain. This is the first report on isobutanol production by an engineered acetogen biocatalyst suitable for commercial manufacturing of this chemical/fuel using continuous fermentation of CO2/H2 blend thus contributing to the reversal of global warming.

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Acknowledgments

The research was supported by the angel funds of MT family friends. Biocatalyst Gurus, Inc. is the sole distributor of the electroporation and electrofusion equipment, custom multi-fermentor modules and MT254UVnator: www.syngasbiofuelsenergy.com.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Syngas Biofuels Energy, Inc., P.O. Box 300819, Houston, TX, 77230, USA

    Michael Tyurin

  2. Prosp. Andropova 19, kv. 27, 115470, Moscow, Russia

    Eugene Gak & Michael Kiriukhin

Authors
  1. Eugene Gak
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  2. Michael Tyurin
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  3. Michael Kiriukhin
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Correspondence to Michael Tyurin.

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The authors have made equal contribution to the reported project.

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Gak, E., Tyurin, M. & Kiriukhin, M. Genome tailoring powered production of isobutanol in continuous CO2/H2 blend fermentation using engineered acetogen biocatalyst. J Ind Microbiol Biotechnol 41, 763–781 (2014). https://doi.org/10.1007/s10295-014-1416-5

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  • DOI: https://doi.org/10.1007/s10295-014-1416-5

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