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Gene modification of Escherichia coli and incorporation of process control to decrease acetate accumulation and increase ʟ-tryptophan production

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Abstract

Acetate is a primary inhibitory metabolite in cultures of Escherichia coli, and the production of both biomass and desired products are increased by reducing the accumulation of acetate. In this study, the accumulation of acetate during ʟ-tryptophan production was decreased by genetic modification of ʟ-tryptophan-producing strain (BCTRP) and optimization of the fermentation process. The mutant (BCTRPG), which has a deletion of the integral membrane permease IICBGlc (ptsG), produces a higher concentration of ʟ-tryptophan than mutants with deletions of either phosphate acetyltransferase (pta) or pta–ptsG, due to the low accumulation of acetate and other byproducts, as well as high biomass production. The appropriate dissolved oxygen (DO) level, glucose feeding mode, and pH control strategy were applied to ʟ-tryptophan production using the BCTRPG mutant. The BCTRPG strain with optimized conditions resulted in a reduction in acetate accumulation (71.08% reduction to 0.72 g/L) and an increase in ʟ-tryptophan production (35.81% increase to 17.14 g/L) compared with the BCTRP strain in the original culture condition. Meanwhile, an analysis of the metabolic flux distribution indicated that the acetate synthesis flux decreased from 19.2% (original conditions) to 8.4% (optimized conditions), and the flux of tryptophan formation with the optimized conditions was 18.5%, which was 1.89 times higher than under the original conditions. This study provided the theoretical foundation and technical support for high-level industrialization production of ʟ-tryptophan.

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Acknowledgments

This work was supported by the Doctoral Program Foundation of Shandong Binzhou Animal Science and Veterinary Medicine Academy (BS201402), Development of Science and Technology Plan Program of Binzhou (2015ZC0107), and Agricultural Science and Technology Innovation Project of Shandong Academy of Agricultural Sciences (CXGC2017B02).

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Author notes

  1. Na Lu and Bin Zhang contributed equally to this work.

Authors and Affiliations

  1. School of Life Science, Baicheng Normal University, Baicheng, 137000, People’s Republic of China

    Na Lu

  2. Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou, 256600, People’s Republic of China

    Na Lu, Bin Zhang, Likun Cheng, Songlin Zhang, Shijun Fu, Yueqiang Xiao & Hongyan Liu

  3. College of Biotechnology, Henan University of Technology, Zhengzhou, 450001, People’s Republic of China

    Bin Zhang & Jinshui Wang

  4. Binzhou Marine Environmental Monitoring Station of Shandong Province, Binzhou, 256600, People’s Republic of China

    Hongyan Liu

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  1. Na Lu
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  2. Bin Zhang
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  3. Likun Cheng
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  4. Jinshui Wang
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  5. Songlin Zhang
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  6. Shijun Fu
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  7. Yueqiang Xiao
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  8. Hongyan Liu
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Corresponding authors

Correspondence to Likun Cheng, Jinshui Wang or Hongyan Liu.

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Funding

This study was funded by the Doctoral Program Foundation of Shandong Binzhou Animal Science and Veterinary Medicine Academy (BS201402), Development of Science and Technology Plan Program of Binzhou (2015ZC0107), and Agricultural Science and Technology Innovation Project of Shandong Academy of Agricultural Sciences (CXGC2017B02).

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The authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Lu, N., Zhang, B., Cheng, L. et al. Gene modification of Escherichia coli and incorporation of process control to decrease acetate accumulation and increase ʟ-tryptophan production. Ann Microbiol 67, 567–576 (2017). https://doi.org/10.1007/s13213-017-1289-8

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  • DOI: https://doi.org/10.1007/s13213-017-1289-8

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