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

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).

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Improving lysine production by Corynebacterium glutamicum through DNA microarray-based identification of novel target genes

Applied Microbiology and Biotechnology ◽

10.1007/s00253-007-0916-x ◽

2007 ◽

Vol 76 (3) ◽

pp. 677-689 ◽

Cited By ~ 48

Author(s):

Georg Sindelar ◽

Volker F. Wendisch

Keyword(s):

Corynebacterium Glutamicum ◽

Dna Microarray ◽

Target Genes ◽

Lysine Production ◽

Novel Target

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Direct l-lysine production from cellobiose by Corynebacterium glutamicum displaying beta-glucosidase on its cell surface

Applied Microbiology and Biotechnology ◽

10.1007/s00253-013-5009-4 ◽

2013 ◽

Vol 97 (16) ◽

pp. 7165-7172 ◽

Cited By ~ 39

Author(s):

Noriko Adachi ◽

Chihiro Takahashi ◽

Naoko Ono-Murota ◽

Rie Yamaguchi ◽

Tsutomu Tanaka ◽

...

Keyword(s):

Cell Surface ◽

Corynebacterium Glutamicum ◽

Lysine Production ◽

Beta Glucosidase

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

Applied and Environmental Microbiology ◽

10.1128/aem.70.12.7277-7287.2004 ◽

2004 ◽

Vol 70 (12) ◽

pp. 7277-7287 ◽

Cited By ~ 71

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.

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