Overexpression of alpha-ketoglutarate dehydrogenase in Yarrowia lipolytica and its effect on production of organic acids.
The yeast Yarrowia lipolytica is one of the most intensively studied "non-conventional" yeast species. Its ability to secrete various organic acids, like pyruvic (PA), citric, isocitric, and alpha-ketoglutaric (KGA) acid, in large amounts is of interest for biotechnological applications. We have studied the effect of the alpha-ketoglutarate dehydrogenase (KGDH) complex on the production process of KGA. Being well studied in Saccharomyces cerevisiae this enzyme complex consists of three subunits: alpha-ketoglutarate dehydrogenase, dihydrolipoyl transsuccinylase, and lipoamide dehydrogenase. Here we report the effect of overexpression of these subunits encoding genes and resulting increase of specific KGDH activity on organic acid production under several conditions of growth limitation and an excess of carbon source in Y. lipolytica. The constructed strain containing multiple copies of all three KGDH genes showed a reduced production of KGA and an elevated production of PA under conditions of KGA production. However, an increased activity of the KGDH complex had no influence on organic acid production under citric acid production conditions.
Institut für Mikrobiologie, Technische Universität Dresden, 01062 Dresden, Germany. martina.holz@tu-dresden.de
Martina
M
Holz
Christina
C
Otto
Anne
A
Kretzschmar
Venelina
V
Yovkova
Andreas
A
Aurich
Markus
M
Pötter
Achim
A
Marx
Gerold
G
Barth
D002264:Carboxylic Acids / Q000378:metabolism*; D015870:Gene Expression; D007655:Ketoglutarate Dehydrogenase Complex / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics
D002264:Carboxylic Acids; D011994:Recombinant Proteins; D007655:Ketoglutarate Dehydrogenase Complex