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Purification and characterization of glycerol‐3‐phosphate dehydrogenase of Saccharomyces cerevisiae
Author(s) -
Albertyn Jacobus,
van Tonder André,
Prior Bernard A.
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)81259-o
Subject(s) - dihydroxyacetone phosphate , dhap , glycerol 3 phosphate dehydrogenase , nad+ kinase , dehydrogenase , chemistry , cofactor , isocitrate dehydrogenase , glycerol , biochemistry , substrate (aquarium) , enzyme , oxidoreductase , chromatography , biology , ecology
The NAD‐dependent glycerol‐3‐phosphate dehydrogenase (glycerol‐3‐phosphate:NAD??? oxidoreductase; EC 1.1.1.8; G3P DHG) was purified 178‐fold to homogeneity from Saceharomyces cerevisiue strain H44‐3D by affinity‐ and ion‐exchange chromatography, SDS‐PAGE indicited that the enzyme had a molecular mass of approximately 42,000 (± 1.000) whereas a molecular mass of 68,000 was observed using gel filtration, implying that the enzyme may exist as a dimer, The pH optimum for the reduction or dihydroxyacetone phosphate (DHAP) was 7.6 and the enzyme had a pt of 7.4. NADPH will not subititute for NADH as coenzyme in the reduction of DHAP. The oxidation of glycerol‐3‐phosphate (G3P) occurs at 3% of the rate of DHAP reduction at pH 7.0. Apparent K m values obtained were 0.023 and 0.54 mM ror NADH and DHAP, respectively, NAD, fructose‐1,6‐bisphosphate (FBP), ATP and ADP inhibited G3P DHG activity, K 1 values obtained for NAD with NADH as variable substrate and FBP with DHAP as variable substrate were 0.93 and 4.8 mM, respectively.