Manipulating the pyruvate dehydrogenase bypass of a multi-vitamin auxotrophic yeast Torulopsis glabrata enhanced pyruvate production

Lett Appl Microbiol. 2004;39(2):199-206. doi: 10.1111/j.1472-765X.2004.01563.x.

Abstract

Aims: To investigate the relationship between the activity of pyruvate dehydrogenase (PDH) bypass and the production of pyruvate of a multi-vitamin auxotrophic yeast Torulopsis glabrata.

Methods and results: Torulopsis glabrata CCTCC M202019, a multi-vitamin auxotrophic yeast that requires acetate for complete growth on glucose minimum medium, was selected after nitrosoguanidine mutagenesis of the parent strain T. glabrata WSH-IP303 screened in previous study [Li et al. (2001) Appl. Microbiol. Biotechnol. 55, 680-685]. Strain CCTCC M202019 produced 21% higher pyruvate than the parent strain and was genetically stable in flask cultures. The activities of the pyruvate metabolism-related enzymes in parent and mutant strains were measured. Compared with the parent strain, the activity of pyruvate decarboxylase (PDC) of the mutant strain CCTCC M202019 decreased by roughly 40%, while the activity of acetyl-CoA synthetase (ACS) of the mutant increased by 103.5 or 57.4%, respectively, in the presence or absence of acetate. Pyruvate production by the mutant strain CCTCC M202019 reached 68.7 g l(-1) at 62 h (yield on glucose of 0.651 g g(-1)) in a 7-l jar fermentor.

Conclusions: The increased pyruvate yield in T. glabrata CCTCC M202019 was due to a balanced manipulation of the PDH bypass, where the shortage of cytoplasmic acetyl-CoA caused by the decreased activity of PDC was properly compensated by the increased activity of ACS.

Significance and impact of the study: Manipulating the PDH bypass may provide an alternative approach to enhance the production of glycolysis-related metabolites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetates / pharmacology
  • Candida glabrata / enzymology*
  • Candida glabrata / genetics
  • Candida glabrata / growth & development
  • Coenzyme A Ligases / genetics
  • Fermentation
  • Mutagenesis
  • Nitrosoguanidines / pharmacology
  • Pyruvate Decarboxylase / genetics
  • Pyruvate Dehydrogenase Complex / genetics
  • Pyruvate Dehydrogenase Complex / metabolism
  • Pyruvic Acid / metabolism*
  • Vitamins / metabolism

Substances

  • Acetates
  • Nitrosoguanidines
  • Pyruvate Dehydrogenase Complex
  • Vitamins
  • Pyruvic Acid
  • Pyruvate Decarboxylase
  • Coenzyme A Ligases
  • acetate-CoA ligase (ADP-forming)