Genetic and biochemical characterization of a short-chain alcohol dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus

Eur J Biochem. 2001 May;268(10):3062-8. doi: 10.1046/j.1432-1327.2001.02201.x.


The gene encoding a short-chain alcohol dehydrogenase, AdhA, has been identified in the hyperthermophilic archaeon Pyrococcus furiosus, as part of an operon that encodes two glycosyl hydrolases, the beta-glucosidase CelB and the endoglucanase LamA. The adhA gene was functionally expressed in Escherichia coli, and AdhA was subsequently purified to homogeneity. The quaternary structure of AdhA is a dimer of identical 26-kDa subunits. AdhA is an NADPH-dependent oxidoreductase that converts alcohols to the corresponding aldehydes/ketones and vice versa, with a rather broad substrate specificity. Maximal specific activities were observed with 2-pentanol (46 U x mg(-1)) and pyruvaldehyde (32 U x mg(-1)) in the oxidative and reductive reaction, respectively. AdhA has an optimal activity at 90 degrees C, at which temperature it has a half life of 22.5 h. The expression of the adhA gene in P. furiosus was demonstrated by activity measurements and immunoblot analysis of cell extracts. A role of this novel type of archaeal alcohol dehydrogenase in carbohydrate fermentation is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Dehydrogenase / chemistry*
  • Alcohol Dehydrogenase / genetics*
  • Alcohol Oxidoreductases / genetics*
  • Amino Acid Sequence
  • Blotting, Western
  • Carbohydrate Metabolism
  • Dimerization
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / metabolism
  • Fermentation
  • Kinetics
  • Molecular Sequence Data
  • Operon
  • Plasmids / metabolism
  • Protein Structure, Quaternary
  • Pyrococcus furiosus / enzymology*
  • Sequence Homology, Amino Acid
  • Substrate Specificity
  • Temperature


  • Alcohol Oxidoreductases
  • Alcohol Dehydrogenase