Secreted glutamic protease rescues aspartic protease Pep deficiency in Aspergillus fumigatus during growth in acidic protein medium

Microbiology (Reading). 2011 May;157(Pt 5):1541-1550. doi: 10.1099/mic.0.048603-0. Epub 2011 Feb 24.

Abstract

In an acidic protein medium Aspergillus fumigatus secretes an aspartic endoprotease (Pep) as well as tripeptidyl-peptidases, a prolyl-peptidase and carboxypeptidases. In addition, LC-MS/MS revealed a novel glutamic protease, AfuGprA, homologous to Aspergillus niger aspergillopepsin II. The importance of AfuGprA in protein digestion was evaluated by deletion of its encoding gene in A. fumigatus wild-type D141 and in a pepΔ mutant. Either A. fumigatus Pep or AfuGprA was shown to be necessary for fungal growth in protein medium at low pH. Exoproteolytic activity is therefore not sufficient for complete protein hydrolysis and fungal growth in a medium containing proteins as the sole nitrogen source. Pep and AfuGprA constitute a pair of endoproteases active at low pH, in analogy to A. fumigatus alkaline protease (Alp) and metalloprotease I (Mep), where at least one of these enzymes is necessary for fungal growth in protein medium at neutral pH. Heterologous expression of AfuGprA in Pichia pastoris showed that the enzyme is synthesized as a preproprotein and that the propeptide is removed through an autoproteolytic reaction at low pH to generate the mature protease. In contrast to A. niger aspergillopepsin II, AfuGprA is a single-chain protein and is structurally more similar to G1 proteases characterized in other non-Aspergillus fungi.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acids / metabolism
  • Amino Acid Sequence
  • Aspartic Acid Proteases / chemistry
  • Aspartic Acid Proteases / genetics
  • Aspartic Acid Proteases / metabolism*
  • Aspergillus fumigatus / enzymology*
  • Aspergillus fumigatus / growth & development*
  • Aspergillus fumigatus / metabolism
  • Culture Media / chemistry*
  • Culture Media / metabolism
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Glutamic Acid / metabolism
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Peptide Hydrolases / chemistry
  • Peptide Hydrolases / genetics
  • Peptide Hydrolases / metabolism*
  • Protein Transport

Substances

  • Acids
  • Culture Media
  • Fungal Proteins
  • Glutamic Acid
  • Aspartic Acid Proteases
  • Peptide Hydrolases