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Biochemistry (Moscow)

, Volume 82, Issue 1, pp 13–23 | Cite as

Glutathione-dependent formaldehyde dehydrogenase homolog from Bacillus subtilis strain R5 is a propanol-preferring alcohol dehydrogenase

  • Raza Ashraf
  • Naeem RashidEmail author
  • Saadia Basheer
  • Iram Aziz
  • Muhammad Akhtar
Article

Abstract

Genome search of Bacillus subtilis revealed the presence of an open reading frame annotated as glutathione-dependent formaldehyde dehydrogenase/alcohol dehydrogenase. The open reading frame consists of 1137 nucleotides corresponding to a polypeptide of 378 amino acids. To examine whether the encoded protein is glutathione-dependent formaldehyde dehydrogenase or alcohol dehydrogenase, we cloned and characterized the gene product. Enzyme activity assays revealed that the enzyme exhibits a metal ion-dependent alcohol dehydrogenase activity but no glutathione-dependent formaldehyde dehydrogenase or aldehyde dismutase activity. Although the protein is of mesophilic origin, optimal temperature for the enzyme activity is 60°C. Thermostability analysis by circular dichroism spectroscopy revealed that the protein is stable up to 60°C. Presence or absence of metal ions in the reaction mixture did not affect the enzyme activity. However, metal ions were necessary at the time of protein production and folding. There was a marked difference in the enzyme activity and CD spectra of the proteins produced in the presence and absence of metal ions. The experimental results obtained in this study demonstrate that the enzyme is a bona-fide alcohol dehydrogenase and not a glutathionedependent formaldehyde dehydrogenase.

Keywords

Bacillus subtilis formaldehyde dehydrogenase alcohol dehydrogenase metal dependent protein folding circular dichroism 

Abbreviations

ADH

alcohol dehydrogenase

CD

circular dichroism

GSH

glutathione

GSH-FDH

glutathione-dependent formaldehyde dehydrogenase

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Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • Raza Ashraf
    • 1
  • Naeem Rashid
    • 1
    Email author
  • Saadia Basheer
    • 1
  • Iram Aziz
    • 1
  • Muhammad Akhtar
    • 2
  1. 1.School of Biological SciencesUniversity of the Punjab, Quaid-e-Azam CampusLahorePakistan
  2. 2.School of Biological SciencesUniversity of SouthamptonSouthamptonUK

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