Purification and Properties of NADP+-linked Glycerol Dehydrogenase from Neurospora crassa

M, Viswanath-Reddy; Pyle, Joseph E.; Howe, H. Branch

doi:10.1099/00221287-107-2-289

Cited by 26 publications

(20 citation statements)

References 19 publications

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“…The M , is by far the lowest found amongst the glycerol dehydrogenases studied. In other fungi, the native enzyme is either a dimer or a tetramer (Sheys & Doughty, 1971 ;Dutler et al, 1977;Viswanath-Reddy et al, 1978).…”

Section: Discussionmentioning

confidence: 99%

Purification and properties of NADP+-dependent glycerol dehydrogenases from Aspergillus nidulans and A. niger

Schuurink

Busink

Hondmann

et al. 1990

Journal of General Microbiology

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Glycerol dehydrogenase, NADP+-specific (EC 1.1.1.72), was purified from mycelium of AspergiZZus nidulans and Aspergillus niger using different purification procedures. Both enzymes had an M, of approximately 38000 and were immunologically cross-reactive, but had different amino acid compositions and isoelectric points. For both enzymes, the substrate specificity was limited to glycerol and erythritol for the oxidative reaction and to dihydroxyacetone (DHA), diacetyl, methylglyoxal, erythrose and D-glyceraldehyde for the reductive reaction. The A. nidulans enzyme had a turnover number twice that of the A. n&er enzyme at pH 6-0, whereas inhibition by NADP+ was less (Ki = 45 ~L M vs 13 p~). It is proposed that both enzymes catalyse in #iuo the reduction of DHA to glycerol and that they are regulated by the anabolic reduction charge.

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Section: Discussionmentioning

confidence: 99%

Purification and properties of NADP+-dependent glycerol dehydrogenases from Aspergillus nidulans and A. niger

Schuurink

Busink

Hondmann

et al. 1990

Journal of General Microbiology

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“…But some of glycerol dehydrogenase, although their activity toward D-erythrose are not known, were reported to have multiple forms. For instance, glycerol dehydrogenase of Neurospora crassa (14) has a molecular weight of 160,000 with a subunit molecular weight of about 43,000. When this preparation was subjected to IEF-PAGE, five active bands with pIs from 3.8 to 5.1 were obtained.…”

Section: Oxidative Reactionmentioning

confidence: 99%

Comparison of three forms of erythrose reductase from an Aureobasidium sp. mutant.

Tokuoka

Ishizuka

Wako

et al. 1992

J. Gen. Appl. Microbiol.

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Two forms of erythrose reductave (ER-1 and ER-2) were purified from an Aureobasidium sp. mutant having high erythritol-producing activity besides the one (ER-3) reported previously. They were purified by chromatofocusing or hydrophobic chromatography after being separated from ER-3 on affinity chromatography. Physicochemical and enzymatic properties of the three forms were compared.Molecular weights were estimated to be 38,000 for ER-1 and 37,000 for ER-2 and ER-3 by SDS-PAGE. Isoelectric points of ER-1, ER-2, and ER-3 were 5.2, 5.0, and 4.8, respectively. All three forms behaved similarly toward pH and temperature, and showed maximum activity at pH 6.5 and 45°C. They showed essentially the same pH-and temperature-stability. D-Erythrose was the best substrate and D-glyceraldehyde, L-erythrulose, and dihydroxyacetone followed. No other aldose and ketose were reduced. There was no significant difference in the substrate specificity of the three forms. Their Km and Vmax for D-erythrose were found to be around 8 mM and 0.38-0.63 ,umol/min/mg, respectively. Significant differences were observed in the behavior of the three forms toward metal ions such as Ag+, Fe3+, and A13+. They showed no oxidative activity at neutral pH but showed some activity at alkaline conditions. Although the activity toward erythritol was much lower (less than 0.1%) compared with the reductive activity, the three forms showed remarkable diversity in pH-activity profile and substrate specificity.Aureobasidium sp. SN-G42 (6), a mutant isolated from Aureobasidium sp. SN-124A, is one of the strains used for commercial production of erythritol. This strain

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“…One pathway involves the use of an NADP + -linked glycerol dehydrogenase, which converts glycerol to glyceraldehyde (Viswanath-Reddy et al, 1978). The second pathway involves the oxidation of glycerol to dihydroxyacetone with the concomitant reduction of NAD + to NADH (May & Sloan, 1981).…”

Section: Introductionmentioning

confidence: 99%

Purification, crystallization and quaternary structure analysis of a glycerol dehydrogenase S305C mutant fromBacillus stearothermophilus

Burke

Ruzheinikov

Sedelnikova

et al. 2001

Acta Crystallogr D Biol Cryst

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Bacillus stearothermophilus glycerol dehydrogenase (GlyDH) is a 39.5 kDa molecular weight metalloenzyme which catalyzes the oxidation of glycerol to dihydroxyacetone with the concomitant reduction of NAD + to NADH. Despite its classi®cation as a member of the`iron-containing' polyol dehydrogenase family, studies on recombinant B. stearothermophilus GlyDH have shown this enzyme to be Zn 2+ -dependent. Crystals of a S305C GlyDH mutant were obtained by the hanging-drop vapour-diffusion method, using ammonium sulfate and PEG 400 as precipitating agents, in the presence and absence of NAD + . The crystals belong to space group I422, with approximate unit-cell parameters a = b = 105, c = 149 A Ê and one subunit in the asymmetric unit, corresponding to a packing density of 2.6 A Ê 3 Da À1 . The crystals diffract X-rays to at least 1.8 A Ê resolution on a synchrotron-radiation source. Determination of the structure will provide insights into the key determinations of catalytic activity of this class of enzymes, for which no structures are currently available.

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Purification and Properties of NADP+-linked Glycerol Dehydrogenase from Neurospora crassa

Cited by 26 publications

References 19 publications

Purification and properties of NADP+-dependent glycerol dehydrogenases from Aspergillus nidulans and A. niger

Purification and properties of NADP+-dependent glycerol dehydrogenases from Aspergillus nidulans and A. niger

Comparison of three forms of erythrose reductase from an Aureobasidium sp. mutant.

Purification, crystallization and quaternary structure analysis of a glycerol dehydrogenase S305C mutant fromBacillus stearothermophilus

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