Extremophiles

, Volume 21, Issue 6, pp 1101–1110 | Cite as

Pcal_1311, an alcohol dehydrogenase homologue from Pyrobaculum calidifontis, displays NADH-dependent high aldehyde reductase activity

  • Raza Ashraf
  • Naeem Rashid
  • Tamotsu Kanai
  • Tadayuki Imanaka
  • Muhammad Akhtar
Original Paper
  • 120 Downloads

Abstract

Genome sequence of Pyrobaculum calidifontis, a hyperthermophilic archaeon, harbors three open-reading frames annotated as alcohol dehydrogenases. One of them, Pcal_1311, does not display a significantly high homology with any of the characterized alcohol dehydrogenases. Highest homology of 38% was found with the characterized counterpart from Geobacillus stearothermophilus. To examine the biochemical properties of Pcal_1311, we have cloned and functionally expressed the gene in Escherichia coli. Purified recombinant Pcal_1311 catalyzed the NAD(H)-dependent oxidation of various alcohols and reduction of aldehydes, with a marked preference for substrates with functional group at the terminal carbon. Highest activity for the oxidation reaction (3 μmol min−1 mg−1) was found with 1,4-butanediol and for the reduction reaction (150 μmol min−1 mg−1) with glutaraldehyde. Both the oxidation and reduction activities increased with the increase in temperature up to 80 °C. Recombinant Pcal_1311 was highly stable and retained more than 90% activity even after incubation of 180 min at 90 °C. In addition to the thermostabilty, Pcal_1311 was highly stable in the presence of known denaturants including urea and guanidine hydrochloride. The high stability, particularly thermostability, and the NADH-dependent aldehyde reduction activity make Pcal_1311 a unique member in the alcohol dehydrogenase family.

Keywords

Pyrobaculum calidifontis Alcohol dehydrogenase Aldehyde reductase Denaturant tolerance Thermostable Circular dichroism 

Supplementary material

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Supplementary material 1 (PPTX 144 kb)
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Supplementary material 5 (PPTX 111 kb)

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Copyright information

© Springer Japan KK 2017

Authors and Affiliations

  • Raza Ashraf
    • 1
  • Naeem Rashid
    • 1
  • Tamotsu Kanai
    • 2
  • Tadayuki Imanaka
    • 3
  • Muhammad Akhtar
    • 1
    • 4
  1. 1.School of Biological SciencesUniversity of the PunjabLahorePakistan
  2. 2.Department of Synthetic Chemistry and Biological Chemistry, Graduate School of EngineeringKyoto UniversityKyotoJapan
  3. 3.The Research Organization of Science and TechnologyRitsumeikan UniversityKusatsuJapan
  4. 4.School of Biological SciencesUniversity of SouthamptonSouthamptonUK

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