Boosting Anaplerotic Reactions by Pyruvate Kinase Gene Deletion and Phosphoenolpyruvate Carboxylase Desensitization for Glutamic Acid and Lysine Production in Corynebacterium glutamicum

2016 ◽  
pp. 181-198 ◽  
Author(s):  
Atsushi Yokota ◽  
Kazunori Sawada ◽  
Masaru Wada
Keyword(s):  
Glutamic Acid ◽  
Corynebacterium Glutamicum ◽  
Pyruvate Kinase ◽  
Phosphoenolpyruvate Carboxylase ◽  
Gene Deletion ◽  
Kinase Gene ◽  
Lysine Production ◽  
Anaplerotic Reactions

Importance of Phosphoenolpyruvate Carboxylase of Corynebacterium glutamicum during the Temperature Triggered Glutamic Acid Fermentation

1999 ◽  
Vol 1 (4) ◽  
pp. 334-343 ◽  
Author(s):  
S. Delaunay ◽  
D. Uy ◽  
M.F. Baucher ◽  
J.M. Engasser ◽  
A. Guyonvarch ◽  
...  
Keyword(s):  
Glutamic Acid ◽  
Corynebacterium Glutamicum ◽  
Phosphoenolpyruvate Carboxylase ◽  
Acid Fermentation

scholarly journals Deregulation of Feedback Inhibition of Phosphoenolpyruvate Carboxylase for Improved Lysine Production in Corynebacterium glutamicum

2013 ◽  
Vol 80 (4) ◽  
pp. 1388-1393 ◽  
Author(s):  
Zhen Chen ◽  
Rajesh Reddy Bommareddy ◽  
Doinita Frank ◽  
Sugima Rappert ◽  
An-Ping Zeng
Keyword(s):  
Corynebacterium Glutamicum ◽  
Mutant Strain ◽  
Phosphoenolpyruvate Carboxylase ◽  
Metabolic Flux ◽  
Feedback Inhibition ◽  
Allosteric Regulation ◽  
Flux Analysis ◽  
Lysine Production ◽  
Content Type ◽  
Metabolic Flux Distribution

ABSTRACTAllosteric regulation of phosphoenolpyruvate carboxylase (PEPC) controls the metabolic flux distribution of anaplerotic pathways. In this study, the feedback inhibition ofCorynebacterium glutamicumPEPC was rationally deregulated, and its effect on metabolic flux redistribution was evaluated. Based on rational protein design, six PEPC mutants were designed, and all of them showed significantly reduced sensitivity toward aspartate and malate inhibition. Introducing one of the point mutations (N917G) into theppcgene, encoding PEPC of the lysine-producing strainC. glutamicumLC298, resulted in ∼37% improved lysine production.In vitroenzyme assays and13C-based metabolic flux analysis showed ca. 20 and 30% increases in the PEPC activity and corresponding flux, respectively, in the mutant strain. Higher demand for NADPH in the mutant strain increased the flux toward pentose phosphate pathway, which increased the supply of NADPH for enhanced lysine production. The present study highlights the importance of allosteric regulation on the flux control of central metabolism. The strategy described here can also be implemented to improve other oxaloacetate-derived products.

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).

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