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

Purification, crystallization and quaternary structure analysis of a glycerol dehydrogenase S305C mutant from Bacillus stearothermophilus
Purification, crystallization and quaternary structure analysis of a glycerol dehydrogenase S305C mutant from Bacillus stearothermophilus
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 classification as a member of the `iron-containing' polyol dehydrogenase family, studies on recombinant B. stearothermophilus GlyDH have shown this enzyme to be Zn2+-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 Å and one subunit in the asymmetric unit, corresponding to a packing density of 2.6 Å3 Da-1. The crystals diffract X-rays to at least 1.8 Å 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.
glycerol dehydrogenase, metalloenzyme, bacillus stearothermophilus
0907-4449
165-167
Burke, J.
fa64b23c-c20c-421b-a91f-2c1342eb5177
Ruzheinikov, S.N.
c6d2c62c-f7dc-40b9-8967-9ce0750204ef
Sedelnikova, S.
f766d5be-b9c1-4037-96cd-c2027394b3da
Baker, P.J.
5c7a5426-fe20-432d-b36a-d2afce2d044f
Holmes, D.
0bd269a5-3bab-477c-8bb0-a8711a0aa14b
Muir, N.M.
5ac70d29-d419-4cd8-b981-6ec09c83f0a0
Gore, M.G.
7bd6db4b-c5a2-4206-8666-b92208ba7979
Rice, D.W.
b9850307-dc18-4a15-8350-5969b097cadc
Burke, J.
fa64b23c-c20c-421b-a91f-2c1342eb5177
Ruzheinikov, S.N.
c6d2c62c-f7dc-40b9-8967-9ce0750204ef
Sedelnikova, S.
f766d5be-b9c1-4037-96cd-c2027394b3da
Baker, P.J.
5c7a5426-fe20-432d-b36a-d2afce2d044f
Holmes, D.
0bd269a5-3bab-477c-8bb0-a8711a0aa14b
Muir, N.M.
5ac70d29-d419-4cd8-b981-6ec09c83f0a0
Gore, M.G.
7bd6db4b-c5a2-4206-8666-b92208ba7979
Rice, D.W.
b9850307-dc18-4a15-8350-5969b097cadc

Burke, J., Ruzheinikov, S.N., Sedelnikova, S., Baker, P.J., Holmes, D., Muir, N.M., Gore, M.G. and Rice, D.W. (2001) Purification, crystallization and quaternary structure analysis of a glycerol dehydrogenase S305C mutant from Bacillus stearothermophilus. Acta Crystallographica Section D: Biological Crystallography, 57 (1), 165-167. (doi:10.1107/S0907444900014918).

Record type: Article

Abstract

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 classification as a member of the `iron-containing' polyol dehydrogenase family, studies on recombinant B. stearothermophilus GlyDH have shown this enzyme to be Zn2+-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 Å and one subunit in the asymmetric unit, corresponding to a packing density of 2.6 Å3 Da-1. The crystals diffract X-rays to at least 1.8 Å 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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More information

Published date: 1 January 2001
Keywords: glycerol dehydrogenase, metalloenzyme, bacillus stearothermophilus

Identifiers

Local EPrints ID: 56470
URI: https://eprints.soton.ac.uk/id/eprint/56470
ISSN: 0907-4449
PURE UUID: 4b700226-240c-42d7-957e-3a38ef314154

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Date deposited: 06 Aug 2008
Last modified: 13 Mar 2019 20:35

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