A Corynebacterium glutamicum strain with inactivated pyruvate dehydrogenase complex and a deletion of the gene encoding the pyruvate:quinone oxidoreductase produces about 19 mM L: -valine, 28 mM L: -alanine and about 55 mM pyruvate from 150 mM glucose. Based on this double mutant C. glutamicum △aceE △pqo, we engineered C. glutamicum for efficient production of pyruvate from glucose by additional deletion of the ldhA gene encoding NAD(+)-dependent L: -lactate dehydrogenase (LdhA) and introduction of a attenuated variant of the acetohydroxyacid synthase (△C-T IlvN). The latter modification abolished overflow metabolism towards L: -valine and shifted the product spectrum to pyruvate production. In shake flasks, the resulting strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN produced about 190 mM pyruvate with a Y (P/S) of 1.36 mol per mol of glucose; however, it still secreted significant amounts of L: -alanine. Additional deletion of genes encoding the transaminases AlaT and AvtA reduced L: -alanine formation by about 50\%. In fed-batch fermentations at high cell densities with adjusted oxygen supply during growth and production (0-5\% dissolved oxygen), the newly constructed strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN △alaT △avtA produced more than 500 mM pyruvate with a maximum yield of 0.97 mol per mole of glucose and a productivity of 0.92 mmol g ((CDW)) (-1) h(-1) (i.e., 0.08 g g((CDW)) (-1) h(-1)) in the production phase.
%0 Journal Article
%1 wieschalka_engineering_2012
%A Wieschalka, Stefan
%A Blombach, Bastian
%A Eikmanns, Bernhard J.
%D 2012
%J Appl. Microbiol. Biotechnol.
%K Acid, Corynebacterium Deletion, Engineering Expression, Gene Glucose, Metabolic Networks Pathways, Pyruvic Valine, and glutamicum, myown
%N 2
%P 449--459
%R 10.1007/s00253-011-3843-9
%T Engineering Corynebacterium glutamicum for the production of pyruvate
%V 94
%X A Corynebacterium glutamicum strain with inactivated pyruvate dehydrogenase complex and a deletion of the gene encoding the pyruvate:quinone oxidoreductase produces about 19 mM L: -valine, 28 mM L: -alanine and about 55 mM pyruvate from 150 mM glucose. Based on this double mutant C. glutamicum △aceE △pqo, we engineered C. glutamicum for efficient production of pyruvate from glucose by additional deletion of the ldhA gene encoding NAD(+)-dependent L: -lactate dehydrogenase (LdhA) and introduction of a attenuated variant of the acetohydroxyacid synthase (△C-T IlvN). The latter modification abolished overflow metabolism towards L: -valine and shifted the product spectrum to pyruvate production. In shake flasks, the resulting strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN produced about 190 mM pyruvate with a Y (P/S) of 1.36 mol per mol of glucose; however, it still secreted significant amounts of L: -alanine. Additional deletion of genes encoding the transaminases AlaT and AvtA reduced L: -alanine formation by about 50\%. In fed-batch fermentations at high cell densities with adjusted oxygen supply during growth and production (0-5\% dissolved oxygen), the newly constructed strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN △alaT △avtA produced more than 500 mM pyruvate with a maximum yield of 0.97 mol per mole of glucose and a productivity of 0.92 mmol g ((CDW)) (-1) h(-1) (i.e., 0.08 g g((CDW)) (-1) h(-1)) in the production phase.
@article{wieschalka_engineering_2012,
abstract = {A Corynebacterium glutamicum strain with inactivated pyruvate dehydrogenase complex and a deletion of the gene encoding the pyruvate:quinone oxidoreductase produces about 19 mM L: -valine, 28 mM L: -alanine and about 55 mM pyruvate from 150 mM glucose. Based on this double mutant C. glutamicum △aceE △pqo, we engineered C. glutamicum for efficient production of pyruvate from glucose by additional deletion of the ldhA gene encoding NAD(+)-dependent L: -lactate dehydrogenase (LdhA) and introduction of a attenuated variant of the acetohydroxyacid synthase (△C-T IlvN). The latter modification abolished overflow metabolism towards L: -valine and shifted the product spectrum to pyruvate production. In shake flasks, the resulting strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN produced about 190 mM pyruvate with a Y (P/S) of 1.36 mol per mol of glucose; however, it still secreted significant amounts of L: -alanine. Additional deletion of genes encoding the transaminases AlaT and AvtA reduced L: -alanine formation by about 50\%. In fed-batch fermentations at high cell densities with adjusted oxygen supply during growth and production (0-5\% dissolved oxygen), the newly constructed strain C. glutamicum △aceE △pqo △ldhA △C-T ilvN △alaT △avtA produced more than 500 mM pyruvate with a maximum yield of 0.97 mol per mole of glucose and a productivity of 0.92 mmol g ((CDW)) (-1) h(-1) (i.e., 0.08 g g((CDW)) (-1) h(-1)) in the production phase.},
added-at = {2018-02-09T13:18:17.000+0100},
author = {Wieschalka, Stefan and Blombach, Bastian and Eikmanns, Bernhard J.},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/246bcc08cb2281fd8c7391745cce2ec97/bastian},
doi = {10.1007/s00253-011-3843-9},
interhash = {48b87aa35f4559e7a9ead73d1df42fd1},
intrahash = {46bcc08cb2281fd8c7391745cce2ec97},
issn = {1432-0614},
journal = {Appl. Microbiol. Biotechnol.},
keywords = {Acid, Corynebacterium Deletion, Engineering Expression, Gene Glucose, Metabolic Networks Pathways, Pyruvic Valine, and glutamicum, myown},
language = {eng},
month = apr,
number = 2,
pages = {449--459},
pmid = {22228312},
timestamp = {2018-02-09T12:18:56.000+0100},
title = {Engineering {Corynebacterium} glutamicum for the production of pyruvate},
volume = 94,
year = 2012
}
