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Modification of glycolysis and its effect on the production of l-threonine in Escherichia coli

  • Genetics and Molecular Biology of Industrial Organisms
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

High concentrations of acetate, the main by-product of Escherichia coli (E. coli) high cell density culture, inhibit bacterial growth and l-threonine production. Since metabolic overflux causes acetate accumulation, we attempted to reduce acetate production by redirecting glycolysis flux to the pentose phosphate pathway by deleting the genes encoding phosphofructokinase (pfk) and/or pyruvate kinase (pyk) in an l-threonine-producing strain of E. coli, THRD. pykF, pykA, pfkA, and pfkB deletion mutants produced less acetate (9.44 ± 0.83, 3.86 ± 0.88, 0.30 ± 0.25, and 6.99 ± 0.85 g/l, respectively) than wild-type THRD cultures (19.75 ± 0.93 g/l). THRDΔpykF and THRDΔpykA produced 11.05 and 5.35 % more l-threonine, and achieved a 10.91 and 5.60 % higher yield on glucose, respectively. While THRDΔpfkA grew more slowly and produced less l-threonine than THRD, THRDΔpfkB produced levels of l-threonine (102.28 ± 2.80 g/l) and a yield on glucose (0.34 g/g) similar to that of THRD. The dual deletion mutant THRDΔpfkBΔpykF also achieved low acetate (7.42 ± 0.81 g/l) and high l-threonine yields (111.37 ± 2.71 g/l). The level of NADPH in THRDΔpfkA cultures was depressed, whereas all other mutants produced more NADPH than THRD did. These results demonstrated that modification of glycolysis in E. coli THRD reduced acetate production and increased accumulation of l-threonine.

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Acknowledgments

This research was supported by the National High Technology Research and Development Program (2012AA022102 and 2012AA02A703) and the National Basic Research Program of China (2013CB733901).

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Authors and Affiliations

  1. National and Local United Engineering Lab of Metabolic Control Fermentation Technology, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China

    Xixian Xie, Yuan Liang, Hongliang Liu, Yuan Liu, Qingyang Xu, Chenglin Zhang & Ning Chen

  2. Tianjin Engineering Lab of Efficient and Green Amino Acid Manufacture, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China

    Xixian Xie, Yuan Liang, Hongliang Liu, Yuan Liu, Qingyang Xu, Chenglin Zhang & Ning Chen

  3. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China

    Xixian Xie, Yuan Liang, Hongliang Liu, Yuan Liu, Qingyang Xu, Chenglin Zhang & Ning Chen

  4. Metabolic Engineering Laboratory, College of Biotechnology, Tianjin University of Science and Technology, No. 29, 13 Main Street, Tianjin Economic and Technological Development Area, Tianjin, 300457, People’s Republic of China

    Ning Chen

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  1. Xixian Xie
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Correspondence to Ning Chen.

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X. Xie and Y. Liang contributed equally to this work.

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Xie, X., Liang, Y., Liu, H. et al. Modification of glycolysis and its effect on the production of l-threonine in Escherichia coli . J Ind Microbiol Biotechnol 41, 1007–1015 (2014). https://doi.org/10.1007/s10295-014-1436-1

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