Identification in the mold Hypocrea jecorina of the first fungal D-galacturonic acid reductase

Biochemistry. 2005 Aug 23;44(33):11234-40. doi: 10.1021/bi050792f.

Abstract

A d-galacturonic acid reductase and the corresponding gene were identified from the mold Hypocrea jecorina (Trichoderma reesei). We hypothesize that the enzyme is part of a fungal d-galacturonic acid catabolic pathway which has not been described previously and which is distinctly different from the bacterial pathway. H. jecorina grown on d-galacturonic acid exhibits an NADPH-dependent d-galacturonic acid reductase activity. This activity is absent when the mold is grown on other carbon sources. The d-galacturonic acid reductase was purified, and tryptic digests of the purified protein were sequenced. The open reading frame of the corresponding gene was then cloned from a cDNA library. The open reading frame was functionally expressed in the yeast Saccharomyces cerevisiae. A histidine-tagged protein was purified, and the enzyme kinetics were characterized. The enzyme converts in a reversible reaction from d-galacturonic acid and NADPH to l-galactonic acid and NADP. The enzyme also exhibits activity with d-glucuronic acid and dl-glyceraldehyde.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / chemistry*
  • Alcohol Oxidoreductases / genetics
  • Alcohol Oxidoreductases / metabolism
  • Base Sequence
  • Cloning, Molecular
  • Fungal Proteins / chemistry*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Gene Expression
  • Gene Library
  • Hexuronic Acids / chemistry*
  • Hexuronic Acids / metabolism
  • Hypocrea / enzymology*
  • Hypocrea / genetics
  • Molecular Sequence Data
  • NAD (+) and NADP (+) Dependent Alcohol Oxidoreductases
  • NADP / chemistry
  • NADP / metabolism
  • Oxidation-Reduction
  • Saccharomyces cerevisiae / genetics
  • Substrate Specificity / physiology
  • Sugar Acids / chemistry
  • Sugar Acids / metabolism

Substances

  • Fungal Proteins
  • Hexuronic Acids
  • Sugar Acids
  • galactonic acid
  • galacturonic acid
  • NADP
  • Alcohol Oxidoreductases
  • NAD (+) and NADP (+) Dependent Alcohol Oxidoreductases
  • D-galacturonate reductase