Metabolic fluxes in Corynebacterium glutamicum during lysine production with sucrose as carbon source.
Metabolic fluxes in the central metabolism were determined for lysine-producing Corynebacterium glutamicum ATCC 21526 with sucrose as a carbon source, providing an insight into molasses-based industrial production processes with this organism. For this purpose, 13C metabolic flux analysis with parallel studies on [1-(13C)Fru]sucrose, [1-(13C)Glc]sucrose, and [13C6Fru]sucrose was carried out. C. glutamicum directed 27.4% of sucrose toward extracellular lysine. The strain exhibited a relatively high flux of 55.7% (normalized to an uptake flux of hexose units of 100%) through the pentose phosphate pathway (PPP). The glucose monomer of sucrose was completely channeled into the PPP. After transient efflux, the fructose residue was mainly taken up by the fructose-specific phosphotransferase system (PTS) and entered glycolysis at the level of fructose-1,6-bisphosphate. Glucose-6-phosphate isomerase operated in the gluconeogenetic direction from fructose-6-phosphate to glucose-6-phosphate and supplied additional carbon (7.2%) from the fructose part of the substrate toward the PPP. This involved supply of fructose-6-phosphate from the fructose part of sucrose either by PTS(Man) or by fructose-1,6-bisphosphatase. C. glutamicum further exhibited a high tricarboxylic acid (TCA) cycle flux of 78.2%. Isocitrate dehydrogenase therefore significantly contributed to the total NADPH supply of 190%. The demands for lysine (110%) and anabolism (32%) were lower than the supply, resulting in an apparent NADPH excess. The high TCA cycle flux and the significant secretion of dihydroxyacetone and glycerol display interesting targets to be approached by genetic engineers for optimization of the strain investigated.
Wittmann
Christoph
C
Biochemical Engineering, Saarland University, Saarbrücken, Germany. c.wittmann@mx.uni-saarland.de
Kiefer
Patrick
P
Zelder
Oskar
O
D002247:Carbon Isotopes / Q000378:metabolism; D002952:Citric Acid Cycle; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D003470:Culture Media; D008401:Gas Chromatography-Mass Spectrometry; D006019:Glycolysis; D008239:Lysine / Q000096:biosynthesis*; D009154:Mutation; D010427:Pentose Phosphate Pathway; D013395:Sucrose / Q000378:metabolism*
D002247:Carbon Isotopes; D003470:Culture Media; D013395:Sucrose; D008239:Lysine