A novel gnd mutation leading to increased L-lysine production in Corynebacterium glutamicum.
Toward more efficient L-lysine production, we have been challenging genome-based strain breeding by the approach of assembling only relevant mutations in a single wild-type background. Following the creation of a new L-lysine producer Corynebacterium glutamicum AHP-3 that carried three useful mutations (lysC311, hom59, and pyc458) on the relevant downstream pathways, we shifted our target to the pentose phosphate pathway. Comparative genomic analysis for the pathway between a classically derived L-lysine producer and its parental wild-type identified several mutations. Among these mutations, a Ser-361-->Phe mutation in the 6-phosphogluconate dehydrogenase gene (gnd) was defined as a useful mutation for L-lysine production. Introduction of the gnd mutation into strain AHP-3 by allelic replacement led to approximately 15% increased L-lysine production. Enzymatic analysis revealed that the mutant enzyme was less sensitive than the wild-type enzyme to allosteric inhibition by intracellular metabolites, such as fructose 1,6-bisphosphate, D-glyceraldehyde 3-phosphate, phosphoribosyl pyrophosphate, ATP, and NADPH, which were known to inhibit this enzyme. Isotope-based metabolic flux analysis demonstrated that the gnd mutation resulted in 8% increased carbon flux through the pentose phosphate pathway during L-lysine production. These results indicate that the gnd mutation is responsible for diminished allosteric regulation and contributes to redirection of more carbon to the pentose phosphate pathway that was identified as the primary source for NADPH essential for L-lysine biosynthesis, thereby leading to improved product formation.
Ohnishi
Junko
J
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Asahi-machi, Machida, Tokyo 194-8533, Japan.
Katahira
Ritsuko
R
Mitsuhashi
Satoshi
S
Kakita
Shingo
S
Ikeda
Masato
M
D003352:Corynebacterium / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D005818:Genetic Engineering; D016680:Genome, Bacterial; D007218:Industrial Microbiology / Q000379:methods; D008239:Lysine / Q000096:biosynthesis*; D009154:Mutation; D010427:Pentose Phosphate Pathway / Q000235:genetics
D008239:Lysine