Universität zu Köln

Krahn, Irene, Bonder, Daniel, Torregrosa-Barragan, Lucia, Stoppel, Dominik, Krause, Jens P., Rosenfeldt, Natalie, Meiswinkel, Tobias M., Seibold, Gerd M., Wendisch, Volker F. ORCID: 0000-0003-3473-0012 and Lindner, Steffen N. (2021). Evolving a New Efficient Mode of Fructose Utilization for Improved Bioproduction in Corynebacterium glutamicum. Front. Bioeng. Biotechnol., 9. LAUSANNE: FRONTIERS MEDIA SA. ISSN 2296-4185

Full text not available from this repository.

Abstract

Fructose utilization in Corynebacterium glutamicum starts with its uptake and concomitant phosphorylation via the phosphotransferase system (PTS) to yield intracellular fructose 1-phosphate, which enters glycolysis upon ATP-dependent phosphorylation to fructose 1,6-bisphosphate by 1-phosphofructokinase. This is known to result in a significantly reduced oxidative pentose phosphate pathway (oxPPP) flux on fructose (similar to 10%) compared to glucose (similar to 60%). Consequently, the biosynthesis of NADPH demanding products, e.g., L-lysine, by C. glutamicum is largely decreased when fructose is the only carbon source. Previous works reported that fructose is partially utilized via the glucose-specific PTS presumably generating fructose 6-phosphate. This closer proximity to the entry point of the oxPPP might increase oxPPP flux and, consequently, NADPH availability. Here, we generated deletion strains lacking either the fructose-specific PTS or 1-phosphofructokinase activity. We used these strains in short-term evolution experiments on fructose minimal medium and isolated mutant strains, which regained the ability of fast growth on fructose as a sole carbon source. In these fructose mutants, the deletion of the glucose-specific PTS as well as the 6-phosphofructokinase gene, abolished growth, unequivocally showing fructose phosphorylation via glucose-specific PTS to fructose 6-phosphate. Gene sequencing revealed three independent amino acid substitutions in PtsG (M260V, M260T, and P318S). These three PtsG variants mediated faster fructose uptake and utilization compared to native PtsG. In-depth analysis of the effects of fructose utilization via these PtsG variants revealed significantly increased ODs, reduced side-product accumulation, and increased L-lysine production by 50%.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Krahn, Irene UNSPECIFIED UNSPECIFIED UNSPECIFIED Bonder, Daniel UNSPECIFIED UNSPECIFIED UNSPECIFIED Torregrosa-Barragan, Lucia UNSPECIFIED UNSPECIFIED UNSPECIFIED Stoppel, Dominik UNSPECIFIED UNSPECIFIED UNSPECIFIED Krause, Jens P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Rosenfeldt, Natalie UNSPECIFIED UNSPECIFIED UNSPECIFIED Meiswinkel, Tobias M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Seibold, Gerd M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Wendisch, Volker F. UNSPECIFIED orcid.org/0000-0003-3473-0012 UNSPECIFIED Lindner, Steffen N. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-586178
DOI:	10.3389/fbioe.2021.669093
Journal or Publication Title:	Front. Bioeng. Biotechnol.
Volume:	9
Date:	2021
Publisher:	FRONTIERS MEDIA SA
Place of Publication:	LAUSANNE
ISSN:	2296-4185
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language L-LYSINE PRODUCTION; ESCHERICHIA-COLI; PHOSPHOTRANSFERASE SYSTEM; CENTRAL METABOLISM; GLUCOSE-UPTAKE; EXPRESSION; FLUX; GROWTH; GLUTAMATE; PHOSPHORYLATION Multiple languages Biotechnology & Applied Microbiology; Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/58617