Metabolic evolution and (13) C flux analysis of a succinate dehydrogenase deficient strain of Yarrowia lipolytica

Biotechnol Bioeng. 2016 Nov;113(11):2425-32. doi: 10.1002/bit.26007. Epub 2016 Jun 23.

Abstract

Bio-based succinic acid production can redirect industrial chemistry processes from using limited hydrocarbons to renewable carbohydrates. A fermentation process that does not require pH-titrating agents will be advantageous to the industry. Previously, a Yarrowia lipolytica strain that was defective for succinate dehydrogenase was constructed and was found to accumulate up to 17.5 g L(-1) of succinic acid when grown on glycerol without buffering. Here, a derivative mutant was isolated that produced 40.5 g L(-1) of succinic acid in 36 h with a yield of 0.32 g g(-1) glycerol. A combination approach of induced mutagenesis and metabolic evolution allowed isolation of another derivative that could utilize glucose efficiently and accumulated 50.2 g L(-1) succinic acid in 54 h with a yield of 0.43 g g(-1) . The parent strain of these isolated mutants was used for [1,6-(13) C2 ]glucose assimilation analysis. At least 35% glucose was estimated to be utilized through the pentose phosphate pathway, while ≥84% succinic acid was formed through the oxidative branch of the tricarboxylic acid cycle. Biotechnol. Bioeng. 2016;113: 2425-2432. © 2016 Wiley Periodicals, Inc.

Keywords: SDH2; Yarrowia lipolytica; low pH; metabolic evolution; succinic acid.

MeSH terms

  • Carbon Isotopes / pharmacokinetics*
  • Glucose / metabolism
  • Metabolic Clearance Rate
  • Metabolic Flux Analysis / methods*
  • Succinate Dehydrogenase / deficiency
  • Succinate Dehydrogenase / genetics
  • Succinate Dehydrogenase / metabolism*
  • Succinic Acid / metabolism*
  • Yarrowia / physiology*

Substances

  • Carbon Isotopes
  • Succinic Acid
  • Succinate Dehydrogenase
  • Glucose