Enhanced l -lysine production in threonine-limited continuous culture of Corynebacterium glutamicum by using gluconate as a secondary carbon source with glucose

1998 ◽  
Vol 49 (1) ◽  
pp. 9-15 ◽  
Author(s):  
H.-W. Lee ◽  
J.-G. Pan ◽  
J.-M. Lebeault
Keyword(s):  
Carbon Source ◽  
Continuous Culture ◽  
Corynebacterium Glutamicum ◽  
Lysine Production ◽  
Secondary Carbon

scholarly journals Metabolic Fluxes in Corynebacterium glutamicum during Lysine Production with Sucrose as Carbon Source

2004 ◽  
Vol 70 (12) ◽  
pp. 7277-7287 ◽  
Author(s):  
Christoph Wittmann ◽  
Patrick Kiefer ◽  
Oskar Zelder
Keyword(s):  
Carbon Source ◽  
Corynebacterium Glutamicum ◽  
Metabolic Flux ◽  
Tca Cycle ◽  
Pentose Phosphate ◽  
Flux Analysis ◽  
Phosphotransferase System ◽  
Lysine Production ◽  
Metabolic Fluxes ◽  
Fructose 1,6 Bisphosphate

ABSTRACT Metabolic fluxes in the central metabolism were determined for lysine-producing Corynebacterium glutamicum ATCC 21526 with sucrose as a carbon source, providing an insight into molasses-based industrial production processes with this organism. For this purpose, 13C metabolic flux analysis with parallel studies on [1-13CFru]sucrose, [1-13CGlc]sucrose, and [13C6 Fru]sucrose was carried out. C. glutamicum directed 27.4% of sucrose toward extracellular lysine. The strain exhibited a relatively high flux of 55.7% (normalized to an uptake flux of hexose units of 100%) through the pentose phosphate pathway (PPP). The glucose monomer of sucrose was completely channeled into the PPP. After transient efflux, the fructose residue was mainly taken up by the fructose-specific phosphotransferase system (PTS) and entered glycolysis at the level of fructose-1,6-bisphosphate. Glucose-6-phosphate isomerase operated in the gluconeogenetic direction from fructose-6-phosphate to glucose-6-phosphate and supplied additional carbon (7.2%) from the fructose part of the substrate toward the PPP. This involved supply of fructose-6-phosphate from the fructose part of sucrose either by PTSMan or by fructose-1,6-bisphosphatase. C. glutamicum further exhibited a high tricarboxylic acid (TCA) cycle flux of 78.2%. Isocitrate dehydrogenase therefore significantly contributed to the total NADPH supply of 190%. The demands for lysine (110%) and anabolism (32%) were lower than the supply, resulting in an apparent NADPH excess. The high TCA cycle flux and the significant secretion of dihydroxyacetone and glycerol display interesting targets to be approached by genetic engineers for optimization of the strain investigated.

l-Lysine production in continuous culture of an l-lysine hyperproducing mutant of Corynebacterium glutamicum

1989 ◽  
Vol 32 (3) ◽  
Author(s):  
Toshihiko Hirao ◽  
Tetsuo Nakano ◽  
Tomoki Azuma ◽  
Masahiro Sugimoto ◽  
Toshihide Nakanishi
Keyword(s):  
Continuous Culture ◽  
Corynebacterium Glutamicum ◽  
Lysine Production

scholarly journals Evolving a new efficient mode of fructose utilization for improved bioproduction in Corynebacterium glutamicum

2021 ◽  
Author(s):  
Irene Krahn ◽  
Daniel Bonder ◽  
Lucia Torregrosa ◽  
Dominik Stoppel ◽  
Jens P. Krause ◽  
...  
Keyword(s):  
Carbon Source ◽  
Corynebacterium Glutamicum ◽  
Pentose Phosphate ◽  
Phosphotransferase System ◽  
Lysine Production ◽  
Depth Analysis ◽  
Mutant Strains ◽  
Phosphofructokinase Activity ◽  
Fructose 1,6 Bisphosphate ◽  
Product Accumulation

AbstractFructose 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 (~10 %) compared to glucose (~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 either lacking in 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, 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 biomass formation, reduced side-product accumulation, and increased L-lysine production by 50 %.

scholarly journals Evolving a New Efficient Mode of Fructose Utilization for Improved Bioproduction in Corynebacterium glutamicum

2021 ◽  
Vol 9 ◽  
Author(s):  
Irene Krahn ◽  
Daniel Bonder ◽  
Lucía Torregrosa-Barragán ◽  
Dominik Stoppel ◽  
Jens P. Krause ◽  
...  
Keyword(s):  
Carbon Source ◽  
Corynebacterium Glutamicum ◽  
Pentose Phosphate ◽  
Phosphotransferase System ◽  
Lysine Production ◽  
Depth Analysis ◽  
Mutant Strains ◽  
Phosphofructokinase Activity ◽  
Fructose 1,6 Bisphosphate ◽  
Product Accumulation

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 (∼10%) compared to glucose (∼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%.

Expression of the NADP+-Dependent Formate Dehydrogenase Gene from Pseudomonas Increases the Lysine Production in Corynebacterium glutamicum

2019 ◽  
Vol 35 (6) ◽  
pp. 21-29
Author(s):  
T.E. Leonova ◽  
T.E. Shustikova ◽  
T.V. Gerasimova ◽  
Т.А. Ivankova ◽  
K.V. Sidorenko Sidorenko ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Formate Dehydrogenase ◽  
Ministry Of Education ◽  
Lysine Production ◽  
Simultaneous Formation ◽  
Resource Center ◽  
Formate Oxidation ◽  
Gene Coding ◽  
Dehydrogenase Gene ◽  
Lysine Synthesis

Thepsefdh_D221Q gene coding for a mutant formate dehydrogenase (PseFDG_D221Q) from Pseudomonas, which catalyzes the formate oxidation with the simultaneous formation of NADPH, has been expressed in the cells of lysine-producing Corynebacterium glutamicum strains. The psefdh_D221Q gene was introduced into С. glutamicum strains as part of an autonomous plasmid or was integrated into the chromosome with simultaneous inactivation of host formate dehydrogenase genes. It was shown that the С. glutamicum strains with NADP+ -dependent formate dehydrogenase have an increased level of L-lysine synthesis in the presence of formate, if their own formate dehydrogenase is inactivated. L-lysine, formate dehydrogenase, NADPH, Corynebacterium glutamicum The work was carried out using the equipment of the Multipurpose Scientific This work was carried out on the equipment of the Multipurpose Scientific Installation of «All-Russian Collection of Industrial Microorganisms», National Bio-Resource Center, NRC «Kurchatov Institute»- GosNIIgenetika. This work was financially supported by the Ministry of Education and Science of Russia (Unique Project Identifier - RFMEFI61017X0011).

Native promoters of Corynebacterium glutamicum and its application in l-lysine production

2017 ◽  
Vol 40 (2) ◽  
pp. 383-391 ◽  
Author(s):  
Xiuling Shang ◽  
Xin Chai ◽  
Xuemei Lu ◽  
Yuan Li ◽  
Yun Zhang ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Lysine Production

Lysine production in continuous culture

1989 ◽  
Vol 20-21 (1) ◽  
pp. 511-528 ◽  
Author(s):  
M. D. Ackerson ◽  
E. C. Clausen ◽  
J. L. Gaddy
Keyword(s):  
Continuous Culture ◽  
Lysine Production
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