Alcohol Oxidase from the Methylotrophic Yeast Ogataea polymorpha: Isolation, Purification, and Bioanalytical Application

Methods Mol Biol. 2021:2280:231-248. doi: 10.1007/978-1-0716-1286-6_15.

Abstract

Alcohol oxidase (EC 1.1.3.13; AOX) is a flavoprotein that catalyzes the oxidation of primary short-chain alcohols to corresponding carbonyl compounds with a concomitant release of hydrogen peroxide. It is a key enzyme of methanol metabolism in methylotrophic yeasts, catalyzing the first step of methanol oxidation to formaldehyde.Here we describe the isolation and purification of AOX from the thermotolerant methylotrophic yeast Ogataea (Hansenula) polymorpha, and using this enzyme in enzymatic assay of ethanol, simultaneous analysis of methanol and formaldehyde, and in construction of amperometric biosensors selective to primary alcohols and formaldehyde.

Keywords: Alcohol oxidase; Amperometric biosensor; Enzymatic assay; Isolation and purification; Methylotrophic yeast; Primary alcohols and formaldehyde.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Alcohol Oxidoreductases / genetics*
  • Alcohol Oxidoreductases / isolation & purification*
  • Alcohol Oxidoreductases / metabolism
  • Batch Cell Culture Techniques
  • Biosensing Techniques
  • Chromatography, Ion Exchange
  • Cloning, Molecular
  • Formaldehyde / analysis
  • Formaldehyde / metabolism
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism
  • Methanol / analysis
  • Methanol / metabolism
  • Saccharomycetales / enzymology
  • Saccharomycetales / genetics
  • Saccharomycetales / growth & development*

Substances

  • Fungal Proteins
  • Formaldehyde
  • Alcohol Oxidoreductases
  • alcohol oxidase
  • Methanol

Supplementary concepts

  • Ogataea angusta