Complete sequence of glycolytic enzymes in the mycorrhizal basidiomycete, Suillus bovinus.
Axenic cultures of Suillus bovinus were cultivated in inorganic liquid medium with glucose as a carbon source at 25 degrees C and continuous supply of oxygen by aeration with compressed air in the dark. Exogenous fructose as sole carbon source yielded about 50% less increase in dry weight than glucose. This resulted from different uptake velocities. Sucrose as sole exogenous carbon source yielded no measurable increase in dry weight. In glucose cultures, activities of all glycolytic enzymes were found. Maximum specific activities varied largely (from about 60 [fructose 6-phosphate kinase] to about 20,000 [triosephosphate isomerase] nmoles.mg protein-1.min-1). Apparent K(m)-values also varied over more than two orders of magnitude (0.035 mM [pyruvate kinase] to 6.16 mM [triosephosphate isomerase]). Fructose 6-phosphate kinase proved to be the fructose 2,6-bisphosphate-regulated type, aldolase the divalent cation-dependent (class II) type and glyceratephosphate mutase the glycerate 2,3-phosphate-independent type of the respective enzymes. Eight of the 10 enzymes exhibited pH-optima between 7.5-8.0. Triosephosphate isomerase and pyruvate kinase showed highest activities at pH 6.5. Regulatory sites within the glycolytic pathway of Suillus bovinus are discussed; fructose 6-phosphate kinase appears to be its main bottle neck.
Kowallik
W
W
Lehrstuhl für Stoffwechselphysiologie, Fakultät für Biologie, Universität Bielefeld, Germany. W.Kowallik@Biologie.Uni-Bielefeld.DE
Thiemann
M
M
Huang
Y
Y
Mutumba
G
G
Beermann
L
L
Broer
D
D
Grotjohann
N
N
D001487:Basidiomycota / Q000201:enzymology* / Q000254:growth & development; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006019:Glycolysis; D006593:Hexokinase / Q000378:metabolism; D010732:Phosphofructokinase-1 / Q000378:metabolism; D010736:Phosphoglycerate Mutase / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism; D014305:Triose-Phosphate Isomerase / Q000378:metabolism
D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D006593:Hexokinase; D010732:Phosphofructokinase-1; D011770:Pyruvate Kinase; D005634:Fructose-Bisphosphate Aldolase; D010751:Phosphopyruvate Hydratase; D014305:Triose-Phosphate Isomerase; D005956:Glucose-6-Phosphate Isomerase; D010736:Phosphoglycerate Mutase; D005947:Glucose