A rapid procedure for the in situ assay of periplasmic, PQQ-dependent methanol dehydrogenase in intact single bacterial colonies

J Microbiol Methods. 2017 Jun:137:46-49. doi: 10.1016/j.mimet.2017.03.013. Epub 2017 Mar 23.

Abstract

Mechanistic details of methanol oxidation catalyzed by the periplasmically-located pyrroloquinoline quinone-dependent methanol dehydrogenase of methylotrophs can be elucidated using site-directed mutants. Here, we present an in situ colony assay of methanol dehydrogenase, which allows robotic screening of large populations of intact small colonies, and regrowth of colonies for subsequent analysis.

Keywords: Dye-linked assay; Methanol dehydrogenase; Methylotrophs; PQQ; Pyrroloquinoline quinone; Robotic screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / analysis*
  • Bacteria / enzymology*
  • Bacteria / growth & development
  • Bacteria / metabolism
  • Bacterial Proteins / metabolism
  • Mass Screening
  • Methanol / metabolism
  • Microbiological Techniques / methods*
  • Mutagenesis, Site-Directed
  • Oxidation-Reduction
  • PQQ Cofactor / metabolism
  • Periplasm / metabolism*
  • Periplasm / microbiology
  • Quinones

Substances

  • Bacterial Proteins
  • Quinones
  • PQQ Cofactor
  • Alcohol Oxidoreductases
  • alcohol dehydrogenase (acceptor)
  • Methanol