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Recombinant Escherichia coli strains deficient in mixed acid fermentation pathways and capable of rapid aerobic growth on glucose with a reduced crabtree effect

Recombinant Escherichia coli strains deficient in mixed acid fermentation pathways and capable of... In this study, we constructed and characterized Escherichia coli strains deficient in mixed acid fermentation pathways, which are capable of rapid aerobic growth on glucose without pronounced bacterial Crabtree effect. The main pathways of production of acetic and lactic acids and ethanol in these strains were inactivated by a deletion of the ackA, pta, poxB, ldhA, and adhE genes. The phosphoenolpyruvate-dependent phosphotransferase system of glucose transport and phosphorylation was inactivated in the strains by a deletion of the ptsG gene. The possibility of alternative transport and phosphorylation of the carbohydrate substrate was ensured in recombinants by constitutive expression of the galP and glk genes, which encode the low-affinity H+-symporter of D-galactose and glucokinase, respectively. The resulting SGM1.0ΔptsG PtacgalP and SGM1.0ΔptsG PLglk PtacgalP strains were capable of rapid aerobic growth in a minimal medium containing 2.0 and 10.0 g/L of glucose and secreted only small amounts of acetic acid and trace amounts of pyruvic acid. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Biochemistry and Microbiology Springer Journals

Recombinant Escherichia coli strains deficient in mixed acid fermentation pathways and capable of rapid aerobic growth on glucose with a reduced crabtree effect

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Publisher
Springer Journals
Copyright
Copyright © 2013 by Pleiades Publishing, Ltd.
Subject
Life Sciences; Biochemistry, general; Microbiology; Medical Microbiology
ISSN
0003-6838
eISSN
1608-3024
DOI
10.1134/S0003683813020105
Publisher site
See Article on Publisher Site

Abstract

In this study, we constructed and characterized Escherichia coli strains deficient in mixed acid fermentation pathways, which are capable of rapid aerobic growth on glucose without pronounced bacterial Crabtree effect. The main pathways of production of acetic and lactic acids and ethanol in these strains were inactivated by a deletion of the ackA, pta, poxB, ldhA, and adhE genes. The phosphoenolpyruvate-dependent phosphotransferase system of glucose transport and phosphorylation was inactivated in the strains by a deletion of the ptsG gene. The possibility of alternative transport and phosphorylation of the carbohydrate substrate was ensured in recombinants by constitutive expression of the galP and glk genes, which encode the low-affinity H+-symporter of D-galactose and glucokinase, respectively. The resulting SGM1.0ΔptsG PtacgalP and SGM1.0ΔptsG PLglk PtacgalP strains were capable of rapid aerobic growth in a minimal medium containing 2.0 and 10.0 g/L of glucose and secreted only small amounts of acetic acid and trace amounts of pyruvic acid.

Journal

Applied Biochemistry and Microbiology – Springer Journals

Published: Feb 26, 2013

References

  • Molecular Cloning: A Laboratory Manual
    Sambrook, J.; Fritsch, E.; Maniatis, T.