Regulation of enzyme activity of alcohol dehydrogenase through its interactions with pyruvate-ferredoxin oxidoreductase in Thermoanaerobacter tengcongensis

Qian Wang, Quanhui Wang, Wei Tong, Xue Bai, Zhen Chen, Jingjing Zhao, Jiyuan Zhang, Siqi Liu

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Alcohol dehydrogenases (ADHs) from thermophilic microorganisms are interesting enzymes that have their potential applications in biotechnology and potentially provide insight into the mechanisms of action of thermo-tolerant proteins. The molecular mechanisms of ADHs under thermal stress in vivo have yet to be explored. Herein, we employed a proteomic strategy to survey the possible interactions of secondary-ADH (2-ADH) with other proteins in Thermoanaerobacter tengcongensis (T. tengcongensis) cultured at 75. °C and found that 2-ADH, pyruvate-ferredoxin oxidoreductase (PFOR) and several glycolytic enzymes coexisted in a protein complex. Using anion exchange chromatography, the elution profile indicated that the native 2-ADH was present in two forms, PFOR-bound and PFOR-free. Immuno-precipitation and pull down analysis further validated the interactions between 2-ADH and PFOR. The kinetic behaviours of 2-ADH either in the recombinant or native form were evaluated with different substrates. The enzyme activity of 2-ADH was inhibited in a non-competitive mode by PFOR, implying the interaction of 2-ADH and PFOR negatively regulated alcohol formation. In T. tengcongensis, PFOR is an enzyme complex located at the upstream of 2-ADH in the alcohol generation pathway. These findings, therefore, offered a plausible mechanism for how alcohol metabolism is regulated by hetero-interactions between 2-ADH and PFOR, especially in anaerobic thermophiles.

Original languageEnglish (US)
Pages (from-to)1018-1023
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume417
Issue number3
DOIs
StatePublished - Jan 20 2012
Externally publishedYes

Keywords

  • Enzymatic activity
  • Protein complex
  • Protein interaction
  • Pyruvate-ferredoxin oxidoreductase (PFOR)
  • Secondary alcohol dehydrogenase (2-ADH)
  • Thermoanaerobacter tengcongensis (T. tengcongensis)

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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