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Current Microbiology

, Volume 62, Issue 3, pp 981–989 | Cite as

Evaluation of Genetic Manipulation Strategies on d-Lactate Production by Escherichia coli

  • Li Zhou
  • Zhi-Rui Zuo
  • Xian-Zhong Chen
  • Dan-Dan Niu
  • Kang-Ming Tian
  • Bernard A. Prior
  • Wei Shen
  • Gui-Yang Shi
  • Suren Singh
  • Zheng-Xiang WangEmail author
Article

Abstract

In order to rationally manipulate the cellular metabolism of Escherichia coli for d-lactate production, single-gene and multiple-gene deletions with mutations in acetate kinase (ackA), phosphotransacetylase (pta), phosphoenolpyruvate synthase (pps), pyruvate formate lyase (pflB), FAD-binding d-lactate dehydrogenase (dld), pyruvate oxidase (poxB), alcohol dehydrogenase (adhE), and fumarate reductase (frdA) were tested for their effects in two-phase fermentations (aerobic growth and oxygen-limited production). Lactate yield and productivity could be improved by single-gene deletions of ackA, pta, pflB, dld, poxB, and frdA in the wild type E. coli strain but were unfavorably affected by deletions of pps and adhE. However, fermentation experiments with multiple-gene mutant strains showed that deletion of pps in addition to ackA-pta deletions had no effect on lactate production, whereas the additional deletion of adhE in E. coli B0013-050 (ackA-pta pps pflB dld poxB) increased lactate yield. Deletion of all eight genes in E. coli B0013 to produce B0013-070 (ackA-pta pps pflB dld poxB adhE frdA) increased lactate yield and productivity by twofold and reduced yields of acetate, succinate, formate, and ethanol by 95, 89, 100, and 93%, respectively. When tested in a bioreactor, E. coli B0013-070 produced 125 g/l d-lactate with an increased oxygen-limited lactate productivity of 0.61 g/g h (2.1-fold greater than E. coli B0013). These kinetic properties of d-lactate production are among the highest reported and the results have revealed which genetic manipulations improved d-lactate production by E. coli.

Keywords

Pyruvate Formate Lyase Lactate Yield Bioreactor Experiment Lactate Fermentation Pyruvate Oxidase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was partly funded by the Sino-South Africa Cooperation Program 2009DFA31300 and the National Natural Science Foundation of China No. 21006039.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Li Zhou
    • 1
  • Zhi-Rui Zuo
    • 1
  • Xian-Zhong Chen
    • 1
    • 2
  • Dan-Dan Niu
    • 1
  • Kang-Ming Tian
    • 1
  • Bernard A. Prior
    • 3
  • Wei Shen
    • 1
    • 2
  • Gui-Yang Shi
    • 1
    • 2
  • Suren Singh
    • 4
  • Zheng-Xiang Wang
    • 1
    • 2
    Email author
  1. 1.Center for Bioresource and Bioenergy, School of BiotechnologyJiangnan UniversityWuxiPeople’s Republic of China
  2. 2.The Key Laboratory of Industrial Biotechnology of Ministry of EducationJiangnan UniversityWuxiPeople’s Republic of China
  3. 3.Department of MicrobiologyUniversity of StellenboschMatielandSouth Africa
  4. 4.Department of Biotechnology and Food Technology, Faculty of Applied SciencesDurban University of TechnologyDurbanSouth Africa

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