caa a22 4500 605506302 CHVBK 20210128100626.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-015-6546-9 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6546-9 Overexpression of the phosphofructokinase encoding gene is crucial for achieving high production of D -lactate in Corynebacterium glutamicum under oxygen deprivation [Elektronische Daten] [Yota Tsuge, Shogo Yamamoto, Naoto Kato, Masako Suda, Alain Vertès, Hideaki Yukawa, Masayuki Inui] We previously reported on the impacts of the overexpression of individual genes of the glycolytic pathway encoding glucokinase (GLK), glyceraldehyde phosphate dehydrogenase (GAPDH), phosphofructokinase (PFK), triosephosphate isomerase (TPI), and bisphosphate aldolase (FBA) on D-lactate productivity in Corynebacterium glutamicum under oxygen-deprived conditions. Searching for synergies, in the current study, we simultaneously overexpressed the five glycolytic genes in a stepwise fashion to evaluate the effect of the cumulative overexpression of glycolytic genes on D-lactate production. Interestingly, the final D-lactate concentration markedly differed depending on whether or not the PFK encoding gene was overexpressed when combined with overexpressing other glycolytic genes. The simultaneous overexpression of the GLK, GAPDH, TPI, and FBA encoding genes led to the highest initial D-lactate concentration at 10h. However, this particular recombinant strain dramatically slowed producing D-lactate when a concentration of 1300mM was reached, typically after 32h. In contrast, the strain overexpressing the PFK encoding gene together with the GLK, GAPDH, TPI, and FBA encoding genes showed 12.7% lower initial D-lactate concentration at 10h than that observed with the strain overexpressing the genes coding for GLK, GAPDH, TPI, and FBA. However, this recombinant strain continued to produce D-lactate after 32h, reaching 2169mM after a mineral salts medium bioprocess incubation period of 80h. These results suggest that overexpression of the PFK encoding gene is essential for achieving high production of D-lactate. Our findings provide interesting options to explore for using C. glutamicum for cost-efficient production of D-lactate at the industrial scale. Springer-Verlag Berlin Heidelberg, 2015 Corynebacterium glutamicum nationallicence Oxygen deprivation nationallicence D -Lactic acid nationallicence Metabolic engineering nationallicence Glycolytic enzyme nationallicence Tsuge Yota Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut Yamamoto Shogo Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut Kato Naoto Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut Suda Masako Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut Vertès Alain Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut Yukawa Hideaki Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut Inui Masayuki Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4679-4689 0175-7598 99:11<4679 2015 99 253 https://doi.org/10.1007/s00253-015-6546-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6546-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tsuge Yota Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut NATIONALLICENCE 700 1- Yamamoto Shogo Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut NATIONALLICENCE 700 1- Kato Naoto Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut NATIONALLICENCE 700 1- Suda Masako Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut NATIONALLICENCE 700 1- Vertès Alain Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut NATIONALLICENCE 700 1- Yukawa Hideaki Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut NATIONALLICENCE 700 1- Inui Masayuki Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, 619-0292, Kizugawa, Kyoto, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4679-4689 0175-7598 99:11<4679 2015 99 253