Production and characterization of a thermostable alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily

M.P. Machielsen, A.R. Uria, S.W.M. Kengen, J. van der Oost

Research output: Contribution to journalArticleAcademicpeer-review

64 Citations (Scopus)

Abstract

The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The enzyme has a monomeric conformation with a molecular mass of 32 kDa. The catalytic activity of the enzyme increases up to 100°C, and a half-life value of 130 min at this temperature indicates its high thermostability. AdhD exhibits a broad substrate specificity with, in general, a preference for the reduction of ketones (pH optimum, 6.1) and the oxidation of secondary alcohols (pH optimum, 8.8). Maximal specific activities were detected with 2,3-butanediol (108.3 U/mg) and diacetyl-acetoin (22.5 U/mg) in the oxidative and reductive reactions, respectively. Gas chromatrography analysis indicated that AdhD produced mainly (S)-2-pentanol (enantiomeric excess, 89%) when 2-pentanone was used as substrate. The physiological role of AdhD is discussed.
LanguageEnglish
Pages233-238
JournalApplied and Environmental Microbiology
Volume72
Issue number1
DOIs
Publication statusPublished - 2006

Fingerprint

Alcohol Dehydrogenase
alcohol dehydrogenase
alcohol
Pyrococcus furiosus
Acetoin
Pentanols
enzyme
butanediol
acetoin
Diacetyl
substrate
diacetyl
gene
Archaea
ketone
Enzymes
substrate specificity
Substrate Specificity
enzymes
ketones

Keywords

  • archaeon pyrococcus-furiosus
  • aldehyde ferredoxin oxidoreductase
  • tungsten-containing enzyme
  • thermococcus strain es-1
  • crystal-structure
  • molecular characterization
  • escherichia-coli
  • proteins
  • binding
  • sulfur

Cite this

@article{23172af8ce7b47a4b7748b4cd8d490cc,
title = "Production and characterization of a thermostable alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily",
abstract = "The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The enzyme has a monomeric conformation with a molecular mass of 32 kDa. The catalytic activity of the enzyme increases up to 100°C, and a half-life value of 130 min at this temperature indicates its high thermostability. AdhD exhibits a broad substrate specificity with, in general, a preference for the reduction of ketones (pH optimum, 6.1) and the oxidation of secondary alcohols (pH optimum, 8.8). Maximal specific activities were detected with 2,3-butanediol (108.3 U/mg) and diacetyl-acetoin (22.5 U/mg) in the oxidative and reductive reactions, respectively. Gas chromatrography analysis indicated that AdhD produced mainly (S)-2-pentanol (enantiomeric excess, 89{\%}) when 2-pentanone was used as substrate. The physiological role of AdhD is discussed.",
keywords = "archaeon pyrococcus-furiosus, aldehyde ferredoxin oxidoreductase, tungsten-containing enzyme, thermococcus strain es-1, crystal-structure, molecular characterization, escherichia-coli, proteins, binding, sulfur",
author = "M.P. Machielsen and A.R. Uria and S.W.M. Kengen and {van der Oost}, J.",
year = "2006",
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language = "English",
volume = "72",
pages = "233--238",
journal = "Applied and Environmental Microbiology",
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publisher = "American Society for Microbiology",
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}

Production and characterization of a thermostable alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily. / Machielsen, M.P.; Uria, A.R.; Kengen, S.W.M.; van der Oost, J.

In: Applied and Environmental Microbiology, Vol. 72, No. 1, 2006, p. 233-238.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Production and characterization of a thermostable alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily

AU - Machielsen, M.P.

AU - Uria, A.R.

AU - Kengen, S.W.M.

AU - van der Oost, J.

PY - 2006

Y1 - 2006

N2 - The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The enzyme has a monomeric conformation with a molecular mass of 32 kDa. The catalytic activity of the enzyme increases up to 100°C, and a half-life value of 130 min at this temperature indicates its high thermostability. AdhD exhibits a broad substrate specificity with, in general, a preference for the reduction of ketones (pH optimum, 6.1) and the oxidation of secondary alcohols (pH optimum, 8.8). Maximal specific activities were detected with 2,3-butanediol (108.3 U/mg) and diacetyl-acetoin (22.5 U/mg) in the oxidative and reductive reactions, respectively. Gas chromatrography analysis indicated that AdhD produced mainly (S)-2-pentanol (enantiomeric excess, 89%) when 2-pentanone was used as substrate. The physiological role of AdhD is discussed.

AB - The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The enzyme has a monomeric conformation with a molecular mass of 32 kDa. The catalytic activity of the enzyme increases up to 100°C, and a half-life value of 130 min at this temperature indicates its high thermostability. AdhD exhibits a broad substrate specificity with, in general, a preference for the reduction of ketones (pH optimum, 6.1) and the oxidation of secondary alcohols (pH optimum, 8.8). Maximal specific activities were detected with 2,3-butanediol (108.3 U/mg) and diacetyl-acetoin (22.5 U/mg) in the oxidative and reductive reactions, respectively. Gas chromatrography analysis indicated that AdhD produced mainly (S)-2-pentanol (enantiomeric excess, 89%) when 2-pentanone was used as substrate. The physiological role of AdhD is discussed.

KW - archaeon pyrococcus-furiosus

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KW - tungsten-containing enzyme

KW - thermococcus strain es-1

KW - crystal-structure

KW - molecular characterization

KW - escherichia-coli

KW - proteins

KW - binding

KW - sulfur

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T2 - Applied and Environmental Microbiology

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SN - 0099-2240

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