Structures of Alcohol Dehydrogenases from Ralstonia and Sphingobium spp. Reveal the Molecular Basis for Their Recognition of ‘Bulky–Bulky’ Ketones

Henry Man; Kinga Kędziora; Justyna Kulig; Annika Frank; Iván Lavandera; Vicente Gotor-Fernández; Dörte Rother; Sam Hart; Johan P. Turkenburg; Gideon Grogan

doi:10.1007/s11244-013-0191-2

Structures of Alcohol Dehydrogenases from Ralstonia and Sphingobium spp. Reveal the Molecular Basis for Their Recognition of ‘Bulky–Bulky’ Ketones

Henry Man, Kinga Kędziora, Justyna Kulig, Annika Frank, Iván Lavandera, Vicente Gotor-Fernández, Dörte Rother, Sam Hart, Johan P. Turkenburg, Gideon Grogan

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

Alcohol dehydrogenases (ADHs) are applied in industrial synthetic chemistry for the production of optically active secondary alcohols. However, the substrate spectrum of many ADHs is narrow, and few, for example, are suitable for the reduction of prochiral ketones in which the carbonyl group is bounded by two bulky and/or hydrophobic groups; so-called ‘bulky–bulky’ ketones. Recently two ADHs, RasADH from Ralstonia sp. DSM 6428, and SyADH from Sphingobium yanoikuyae DSM 6900, have been described, which are distinguished by their ability to accept bulky–bulky ketones as substrates. In order to examine the molecular basis of the recognition of these substrates the structures of the native and NADPH complex of RasADH, and the NADPH complex of SyADH have been determined and refined to resolutions of 1.5, 2.9 and 2.5 Å, respectively. The structures reveal hydrophobic active site tunnels near the surface of the enzymes that are well-suited to the recognition of large hydrophobic substrates, as determined by modelling of the bulky–bulky substrate n-pentyl phenyl ketone. The structures also reveal the bases for NADPH specificity and (S)-stereoselectivity in each of the biocatalysts for n-pentyl phenyl ketone and related substrates.

Original language	English
Pages (from-to)	356-365
Number of pages	10
Journal	TOPICS IN CATALYSIS
Volume	57
Issue number	5
Early online date	13 Nov 2013
DOIs	https://doi.org/10.1007/s11244-013-0191-2
Publication status	Published - Mar 2014

Keywords

Alcohol dehydrogenase
Ketoreductase
NADPH
Oxidoreductase
Enzyme structure

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10.1007/s11244-013-0191-2

Cite this

Man, H., Kędziora, K., Kulig, J., Frank, A., Lavandera, I., Gotor-Fernández, V., Rother, D., Hart, S., Turkenburg, J. P., & Grogan, G. (2014). Structures of Alcohol Dehydrogenases from Ralstonia and Sphingobium spp. Reveal the Molecular Basis for Their Recognition of ‘Bulky–Bulky’ Ketones. TOPICS IN CATALYSIS, 57(5), 356-365. https://doi.org/10.1007/s11244-013-0191-2

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title = "Structures of Alcohol Dehydrogenases from Ralstonia and Sphingobium spp. Reveal the Molecular Basis for Their Recognition of {\textquoteleft}Bulky–Bulky{\textquoteright} Ketones",

abstract = "Alcohol dehydrogenases (ADHs) are applied in industrial synthetic chemistry for the production of optically active secondary alcohols. However, the substrate spectrum of many ADHs is narrow, and few, for example, are suitable for the reduction of prochiral ketones in which the carbonyl group is bounded by two bulky and/or hydrophobic groups; so-called {\textquoteleft}bulky–bulky{\textquoteright} ketones. Recently two ADHs, RasADH from Ralstonia sp. DSM 6428, and SyADH from Sphingobium yanoikuyae DSM 6900, have been described, which are distinguished by their ability to accept bulky–bulky ketones as substrates. In order to examine the molecular basis of the recognition of these substrates the structures of the native and NADPH complex of RasADH, and the NADPH complex of SyADH have been determined and refined to resolutions of 1.5, 2.9 and 2.5 {\AA}, respectively. The structures reveal hydrophobic active site tunnels near the surface of the enzymes that are well-suited to the recognition of large hydrophobic substrates, as determined by modelling of the bulky–bulky substrate n-pentyl phenyl ketone. The structures also reveal the bases for NADPH specificity and (S)-stereoselectivity in each of the biocatalysts for n-pentyl phenyl ketone and related substrates.",

keywords = "Alcohol dehydrogenase, Ketoreductase, NADPH, Oxidoreductase, Enzyme structure",

author = "Henry Man and Kinga K{\c e}dziora and Justyna Kulig and Annika Frank and Iv{\'a}n Lavandera and Vicente Gotor-Fern{\'a}ndez and D{\"o}rte Rother and Sam Hart and Turkenburg, {Johan P.} and Gideon Grogan",

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Man, H, Kędziora, K, Kulig, J, Frank, A, Lavandera, I, Gotor-Fernández, V, Rother, D, Hart, S, Turkenburg, JP & Grogan, G 2014, 'Structures of Alcohol Dehydrogenases from Ralstonia and Sphingobium spp. Reveal the Molecular Basis for Their Recognition of ‘Bulky–Bulky’ Ketones', TOPICS IN CATALYSIS, vol. 57, no. 5, pp. 356-365. https://doi.org/10.1007/s11244-013-0191-2

TY - JOUR

T1 - Structures of Alcohol Dehydrogenases from Ralstonia and Sphingobium spp. Reveal the Molecular Basis for Their Recognition of ‘Bulky–Bulky’ Ketones

AU - Man, Henry

AU - Kędziora, Kinga

AU - Kulig, Justyna

AU - Frank, Annika

AU - Lavandera, Iván

AU - Gotor-Fernández, Vicente

AU - Rother, Dörte

AU - Hart, Sam

AU - Turkenburg, Johan P.

AU - Grogan, Gideon

PY - 2014/3

Y1 - 2014/3

N2 - Alcohol dehydrogenases (ADHs) are applied in industrial synthetic chemistry for the production of optically active secondary alcohols. However, the substrate spectrum of many ADHs is narrow, and few, for example, are suitable for the reduction of prochiral ketones in which the carbonyl group is bounded by two bulky and/or hydrophobic groups; so-called ‘bulky–bulky’ ketones. Recently two ADHs, RasADH from Ralstonia sp. DSM 6428, and SyADH from Sphingobium yanoikuyae DSM 6900, have been described, which are distinguished by their ability to accept bulky–bulky ketones as substrates. In order to examine the molecular basis of the recognition of these substrates the structures of the native and NADPH complex of RasADH, and the NADPH complex of SyADH have been determined and refined to resolutions of 1.5, 2.9 and 2.5 Å, respectively. The structures reveal hydrophobic active site tunnels near the surface of the enzymes that are well-suited to the recognition of large hydrophobic substrates, as determined by modelling of the bulky–bulky substrate n-pentyl phenyl ketone. The structures also reveal the bases for NADPH specificity and (S)-stereoselectivity in each of the biocatalysts for n-pentyl phenyl ketone and related substrates.

AB - Alcohol dehydrogenases (ADHs) are applied in industrial synthetic chemistry for the production of optically active secondary alcohols. However, the substrate spectrum of many ADHs is narrow, and few, for example, are suitable for the reduction of prochiral ketones in which the carbonyl group is bounded by two bulky and/or hydrophobic groups; so-called ‘bulky–bulky’ ketones. Recently two ADHs, RasADH from Ralstonia sp. DSM 6428, and SyADH from Sphingobium yanoikuyae DSM 6900, have been described, which are distinguished by their ability to accept bulky–bulky ketones as substrates. In order to examine the molecular basis of the recognition of these substrates the structures of the native and NADPH complex of RasADH, and the NADPH complex of SyADH have been determined and refined to resolutions of 1.5, 2.9 and 2.5 Å, respectively. The structures reveal hydrophobic active site tunnels near the surface of the enzymes that are well-suited to the recognition of large hydrophobic substrates, as determined by modelling of the bulky–bulky substrate n-pentyl phenyl ketone. The structures also reveal the bases for NADPH specificity and (S)-stereoselectivity in each of the biocatalysts for n-pentyl phenyl ketone and related substrates.

KW - Alcohol dehydrogenase

KW - Ketoreductase

KW - NADPH

KW - Oxidoreductase

KW - Enzyme structure

U2 - 10.1007/s11244-013-0191-2

DO - 10.1007/s11244-013-0191-2

M3 - Article

SN - 1022-5528

VL - 57

SP - 356

EP - 365

JO - TOPICS IN CATALYSIS

JF - TOPICS IN CATALYSIS

IS - 5

ER -