Amplified expression of fructose 1,6-bisphosphatase in Corynebacterium glutamicum increases in vivo flux through the pentose phosphate pathway and lysine production on different carbon sources.
The overexpression of fructose 1,6-bisphosphatase (FBPase) in Corynebacterium glutamicum leads to significant improvement of lysine production on different sugars. Amplified expression of FBPase via the promoter of the gene encoding elongation factor TU (EFTU) increased the lysine yield in the feedback-deregulated lysine-producing strain C. glutamicum lysCfbr by 40% on glucose and 30% on fructose or sucrose. Additionally formation of the by-products glycerol and dihydroxyacetone was significantly reduced in the PEFTUfbp mutant. As revealed by 13C metabolic flux analysis on glucose the overexpression of FBPase causes a redirection of carbon flux from glycolysis toward the pentose phosphate pathway (PPP) and thus leads to increased NADPH supply. Normalized to an uptake flux of glucose of 100%, the relative flux into the PPP was 56% for C. glutamicum lysCfbr PEFTUfbp and 46% for C. glutamicum lysCfbr. The flux for NADPH supply was 180% in the PEFTUfbp strain and only 146% in the parent strain. Amplification of FBPase increases the production of lysine via an increased supply of NADPH. Comparative studies with another mutant containing the sod promoter upstream of the fbp gene indicate that the expression level of FBPase relates to the extent of the metabolic effects. The overexpression of FBPase seems useful for starch- and molasses-based industrial lysine production with C. glutamicum. The redirection of flux toward the PPP should also be interesting for the production of other NADPH-demanding compounds as well as for products directly stemming from the PPP.
Becker
Judith
J
Biochemical Engineering, Saarland University, Im Stadtwald, 66041 Saarbrücken, Germany.
Klopprogge
Corinna
C
Zelder
Oskar
O
Heinzle
Elmar
E
Wittmann
Christoph
C
D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000235:genetics; D017931:DNA Primers; D005246:Feedback; D006597:Fructose-Bisphosphatase / Q000235:genetics*; D005784:Gene Amplification; D007700:Kinetics; D008239:Lysine / Q000096:biosynthesis*; D008954:Models, Biological; D010427:Pentose Phosphate Pathway / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D017931:DNA Primers; D006597:Fructose-Bisphosphatase; D008239:Lysine