title: Phosphotransferase system-independent glucose utilization in corynebacterium glutamicum by inositol permeases and glucokinases.
abstract: Phosphoenolpyruvate-dependent glucose phosphorylation via the phosphotransferase system (PTS) is the major path of glucose uptake in Corynebacterium glutamicum, but some growth from glucose is retained in the absence of the PTS. The growth defect of a deletion mutant lacking the general PTS component HPr in glucose medium could be overcome by suppressor mutations leading to the high expression of inositol utilization genes or by the addition of inositol to the growth medium if a glucokinase is overproduced simultaneously. PTS-independent glucose uptake was shown to require at least one of the inositol transporters IolT1 and IolT2 as a mutant lacking IolT1, IolT2, and the PTS component HPr could not grow with glucose as the sole carbon source. Efficient glucose utilization in the absence of the PTS necessitated the overexpression of a glucokinase gene in addition to either iolT1 or iolT2. IolT1 and IolT2 are low-affinity glucose permeases with K(s) values of 2.8 and 1.9 mM, respectively. As glucose uptake and phosphorylation via the PTS differs from glucose uptake via IolT1 or IolT2 and phosphorylation via glucokinase by the requirement for phosphoenolpyruvate, the roles of the two pathways for l-lysine production were tested. The l-lysine yield by C. glutamicum DM1729, a rationally engineered l-lysine-producing strain, was lower than that by its PTS-deficient derivate DM1729Δhpr, which, however, showed low production rates. The combined overexpression of iolT1 or iolT2 with ppgK, the gene for PolyP/ATP-dependent glucokinase, in DM1729Δhpr enabled l-lysine production as fast as that by the parent strain DM1729 but with 10 to 20% higher l-lysine yield.
authors.affiliation: Genetics of Prokaryotes, Faculty of Biology & CeBiTec, University of Bielefeld, P.O. Box 100131, 33501 Bielefeld, Germany.
authors.forename: Steffen N
authors.initials: SN
authors.lastname: Lindner
authors.forename: Gerd M
authors.initials: GM
authors.lastname: Seibold
authors.forename: Alexander
authors.initials: A
authors.lastname: Henrich
authors.forename: Reinhard
authors.initials: R
authors.lastname: Krämer
authors.forename: Volker F
authors.initials: VF
authors.lastname: Wendisch
mesh_terms: D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism*; D005941:Glucokinase / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D007294:Inositol / Q000378:metabolism; D008239:Lysine / Q000096:biosynthesis; D026901:Membrane Transport Proteins / Q000378:metabolism*; D053858:Metabolic Networks and Pathways / Q000235:genetics; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000172:deficiency / Q000235:genetics
chemical_list: D026901:Membrane Transport Proteins; D007294:Inositol; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; D005941:Glucokinase; D005947:Glucose; D008239:Lysine