Exploring Lysine Riboswitch for Metabolic Flux Control and Improvement of l-Lysine Synthesis in Corynebacterium glutamicum

2015 ◽  
Vol 4 (6) ◽  
pp. 729-734 ◽  
Author(s):  
Li-Bang Zhou ◽  
An-Ping Zeng
Keyword(s):  
Corynebacterium Glutamicum ◽  
Metabolic Flux ◽  
Flux Control ◽  
Lysine Synthesis

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).

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.

scholarly journals The Lys20 homocitrate synthase isoform exerts most of the flux control over the lysine synthesis pathway in Saccharomyces cerevisiae

2011 ◽  
Vol 82 (3) ◽  
pp. 578-590 ◽  
Author(s):  
Héctor Quezada ◽  
Alvaro Marín-Hernández ◽  
Diana Aguilar ◽  
Geovani López ◽  
Juan Carlos Gallardo-Pérez ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Flux Control ◽  
Homocitrate Synthase ◽  
Lysine Synthesis

Pathway identification combining metabolic flux and functional genomics analyses: Acetate and propionate activation by Corynebacterium glutamicum

2009 ◽  
Vol 140 (1-2) ◽  
pp. 75-83 ◽  
Author(s):  
Andrea Veit ◽  
Doris Rittmann ◽  
Tobias Georgi ◽  
Jung-Won Youn ◽  
Bernhard J. Eikmanns ◽  
...  
Keyword(s):  
Functional Genomics ◽  
Corynebacterium Glutamicum ◽  
Metabolic Flux

Impacts of Sodium Citrate on Metabolic Flux Distributions of L-Valine Fermentation

2011 ◽  
Vol 343-344 ◽  
pp. 643-648
Author(s):  
Qing Yang Xu ◽  
Lei Ma ◽  
Xi Xian Xie ◽  
Ning Chen ◽  
Jian Wang
Keyword(s):  
Corynebacterium Glutamicum ◽  
Culture Medium ◽  
Metabolic Flux ◽  
Sodium Citrate ◽  
Flux Distribution ◽  
Metabolic Flux Distribution ◽  
Key Nodes

The effect of sodium citrate on the metabolic flux distributions in the middle and late periods of L-valine production by Corynebacterium glutamicum XV0505 was obtained. It was shown that when sodium citrate (2.0 g/L) was added into the initial fermentation culture medium, the metabolic flux of Embden-Meyerhof-Parnas (EMP) route decreased from 96.43 to 91.13, and the metabolic flux of Hexose Monophophate (HMP) route increased from 3.56 to 8.87, and the metabolic flux flowing to L-alanine and acetate was decreased by 21.1% and 32.4%, respectively. Meanwhile, the metabolic flux of biosynthesis route of L-valine was increased by 10.74%. Therefore, sodium citrate can change the metabolic flux distribution in the key nodes of biosynthesis route of L-valine, decrease the generation of byproducts, and increase the metabolic flux in the biosynthesis route of L-valine.

scholarly journals Enhancing glutamine production by optimising the GS-GOGAT pathway in Corynebacterium glutamicum and NH4+-limited fermentation

2021 ◽  
Author(s):  
Yunpeng Liu ◽  
Lanxiao Li ◽  
Jinduo Wang ◽  
Qingyang Xu
Keyword(s):  
Bacillus Subtilis ◽  
Corynebacterium Glutamicum ◽  
Lactobacillus Acidophilus ◽  
Fermentation Process ◽  
Metabolic Flux ◽  
Feeding Strategies ◽  
Original Strain ◽  
Strong Promoter ◽  
Glutamine Synthase ◽  
Glutamine Production

Abstract: The GS-GOGAT pathway is a key metabolic pathway of glutamate and glutamine. Optimising this pathway, leading to metabolic flux to glutamine, can increase glutamine production and reduce the production of the by-product glutamate. The NH-limited fermentation process limits the concentration of NH to increase the activity of GS and further increase the yield of glutamine. The GS-GOGAT pathway was optimised by knocking out the GOGAT genes NCgl0181 and NCgl0182 and the glutaminase genes NCgl2395 and NCgl2500 and by integrating a copy of the GS gene glnAbsu from Bacillus subtilis and a copy of the glutamine synthase gene glnAlcb from Lactobacillus acidophilus into the genomic NCgl0182 and NCgl2500 sites. Furthermore, the pXT01 plasmid with the strong promoter tuf was used to overexpress glnAbsu and glnAlcb. To obtain an optimal NH-limited fermentation process, the effects of starting feeding with (NH)SO at different times of fermentation and three (NH)SO feeding strategies on glutamine fermentation were studied, and a NH-limited fermentation process that was the most suitable for glutamine fermentation was determined. After optimising the GS-GOGAT pathway, Corynebacterium glutamicum G-6 was subjected to the NH-limited fermentation process to greatly increase the production of glutamine. The yield of glutamine reached 98.7 g/L, which was 104.8% higher than that in the original strain GM34; the content of glutamate reached 4.5 g/L, which then decreased by 85.2%; the GS activity increased significantly, and the sugar-acid conversion rate reached 41.2%.

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