Summary: Glycerol: NAD+ 2-oxidoreductase (glycerol dehydrogenase, EC 1.1.1.6) from Schizosaccharomyces pombe has been purified to homogeneity. The protein has a molecular weight of about 400000; it can be dissociated into identical subunits of molecular weight 47000, and is probably an octamer. The pH optimum for glycerol oxidation is 10·0 or higher and for the reverse reaction is 6·0. Oxidation occurs specifically at C2 of glycerol to produce dihydroxyacetone and not glyceraldehyde. Several diols with hydroxyls on adjacent carbon atoms can be oxidized and corresponding carbonyl compounds reduced, 1,2-propanediol being oxidized 1·6 times more rapidly than glycerol. The forward reaction has a specific requirement for NAD+ as coenzyme whereas NADPH shows about one-third of the activity of NADH for the reverse reaction. The monovalent cations K+ and NH+4 activate the enzyme, while Na+ and Li+ counteract this effect. Some thiol and chelating agents are inhibitory while thiol antagonists, Mn2+, and to a lesser extent Zn2+, stimulate activity. Apparent Km and Vmax values have been determined. The enzyme is similar to but not identical with the glycerol dehydrogenases isolated from Escherichia coli and Klebsiella aerogenes (K. pneumoniae).
HuetingS.,
de LangeT.,
TempestD. W.1978; Properties and regulation of synthesis of the glycerol dehydrogenase present in Klebsiella aerogenes NCTC 418 growing in chemostat culture. FEMS Microbiology Letters 4:185–189
KongY. C.,
MayJ. W.,
MarshallJ. H.1985; Glycerol oxidation and triose reduction by pyridine nucleotide-linked enzymes in the fission yeast Schizosaccharomyces pombe
. Journal of General Microbiology 131:1571–1579
MayJ. W.,
SloanJ.1981; Glycerol utilization by Schizosaccharomyces pombe: dehydrogenation as the initial step. Journal of General Microbiology 123:183–185
McGregorW. G.,
PhillipsJ.,
SuelterC. H.1974; Purification and kinetic characterization of a monovalent cation-activated glycerol dehydrogenase from Aerobacter aerogenes
. Journal of Biological Chemistry 249:3132–3139
PerlinA. S.1962; Trioses, d-, l-, and dl-glyceraldehyde. In Methods in Carbohydrate Chemistry vol 1Analysis and Preparation of Sugars pp 61–63 Edited by
WhistlerR. L.,
WolfromM. L.
New York: Academic Press;
RuchF. E.,
LinE. C. C.,
KowitJ. D.,
TangC. T.,
GoldbergA. L.1980; In vivo inactivation of glycerol dehydrogenase in Klebsiella aerogenes: properties of active and inactivated proteins. Journal of Bacteriology 141:1077–1085
StricklandJ. E.,
MillerO. N.1968; Inhibition of glycerol dehydrogenase from Aerobacter aerogenes by dihydroxyacetone, high ionic strength, and monovalent cations. Biochimica et biophysica acta 159:221–226
TangC. T.,
RuchF. E.,
LinE. C. C.1979; Purification and properties of a nicotinamide adenine dinucleotide-linked dehydrogenase that serves an Escherichia coli mutant for glycerol catabolism. Journal of Bacteriology 140:182–187
TangC. T.,
ForageR. G.,
LinE. C. C.1982a; Immunochemical properties of NAD+-linked glycerol dehydrogenases from Escherichia coli and Klebsiella pneumoniae
. Journal of Bacteriology 152:1169–1174
TangC. T.,
St MartinE. J.,
LinE. C. C.1982b; Derepression of an NAD-linked dehydrogenase that serves an Escherichia coli mutant for growth on glycerol. Journal of Bacteriology 152:1001–1007