Overexpression of the phosphofructokinase encoding gene is crucial for achieving high production of D-lactate in Corynebacterium glutamicum under oxygen deprivation

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• 作者：Yota Tsuge ; Shogo Yamamoto ; Naoto Kato…
• 关键词：Corynebacterium glutamicum ; Oxygen deprivation ; D ; Lactic acid ; Metabolic engineering ; Glycolytic enzyme
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2015
• 出版时间：June 2015
• 年：2015
• 卷：99
• 期：11
• 页码：4679-4689
• 全文大小：485 KB
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  Pearce AK, Crimmins
• 作者单位：Yota Tsuge (1)
  Shogo Yamamoto (1)
  Naoto Kato (1)
  Masako Suda (1)
  Alain A. Vertès (1)
  Hideaki Yukawa (1)
  Masayuki Inui (1) (2)
  
  1. Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, Kizugawa, Kyoto, 619-0292, Japan
  2. Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Nara, 630-0101, Japan
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

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 10?h. However, this particular recombinant strain dramatically slowed producing D-lactate when a concentration of 1300?mM was reached, typically after 32?h. 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 10?h 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 32?h, reaching 2169?mM after a mineral salts medium bioprocess incubation period of 80?h. 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.