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

• Published in: Biotechnology and bioengineering 2016-11, Vol.113 (11), p.2425
• Main Authors: Yuzbashev, Tigran V, Bondarenko, Pavel Yu, Sobolevskaya, Tatiana I, Yuzbasheva, Evgeniya Yu, Laptev, Ivan A, Kachala, Vadim V, Fedorov, Alexander S, Vybornaya, Tatiana V, Larina, Anna S, Sineoky, Sergey P
• Format: Article
• Language: English
• Subjects: 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
• ISSN: 1097-0290
• DOI: 10.1002/bit.26007

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.