Control analysis of the role of triosephosphate isomerase in glucose metabolism in Lactococcus lactis.
Triosephosphate isomerase (TPI), which catalyses the conversion of dihydroxyacetone phosphate (DHAP) to glyceraldehyde-3-phosphate (G3P), was studied for its control on glycolysis and mixed acid production in L. lactis subspecies lactis IL1403 and L. lactis subspecies cremoris MG1363. Strains in which the TPI activity was modulated from 3%-225% (IL1403) or 13%-103% (MG1363) of the wild-type level were constructed by changing the expression of the tpiA gene. The enzyme was found to be present in high excess in the wild-type cells and 10% TPI activity still supported more than 70% of the wild-type glycolytic flux in both strains. Homolactic product formation was preserved throughout the range of TPI activities studied, although a slight increase in the amount of acetate and formate production was observed in the strains with strongly reduced TPI activity for both IL1403 and MG1363. The upstream metabolites glucose-6-phosphate, fructose-1,6-bisphosphate and DHAP in the IL1403 derivatives were essentially unchanged for TPI activities from 26% to 225%. At a TPI activity of 3%, the level of DHAP increased four times. The finding that an increased level of DHAP coincides with an increase in formate production is surprising and indicates that pyruvate formate lyase is not inhibited by DHAP under these conditions.
Solem
C
C
Center for Systems Microbiology, Department of Systems Biology, Technical University of Denmark, Kongens Lyngby, Denmark.
Koebmann
B
B
Jensen
P R
PR
D000123:Acetyltransferases / Q000378:metabolism; D002474:Cell-Free System; D004099:Dihydroxyacetone Phosphate / Q000032:analysis / Q000378:metabolism; D005561:Formates / Q000032:analysis / Q000378:metabolism; D005635:Fructosediphosphates / Q000032:analysis / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D019298:Glucose-6-Phosphate / Q000032:analysis / Q000378:metabolism; D006019:Glycolysis; D013294:Lactococcus lactis / Q000201:enzymology*; D015202:Protein Engineering; D014305:Triose-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism*
D005561:Formates; D005635:Fructosediphosphates; C030544:formic acid; D019298:Glucose-6-Phosphate; D004099:Dihydroxyacetone Phosphate; D000123:Acetyltransferases; C022560:formate C-acetyltransferase; D014305:Triose-Phosphate Isomerase; C029063:fructose-1,6-diphosphate