Production and secretion of Aspergillus nidulans catalase B in filamentous fungi driven by the promoter and signal peptide of the Cladosporium fulvum hydrophobin gene hcf-1

Curr Genet. 2003 Nov;44(3):155-63. doi: 10.1007/s00294-003-0421-4. Epub 2003 Sep 4.

Abstract

We describe here the use of sequences from the hydrophobin gene hcf-1 of Cladosporium fulvum to construct pCatBex, a vector for high-level expression and secretion of CatB, a catalase from Aspergillus nidulans. Transformation of C. fulvum with pCatBex results in a 60-fold increase in the mycelial activity in the fungus and the appearance of up to 5.4 mkat/l of catalase in the growth medium. The levels of catalase in the supernatant increased dramatically following removal of nitrogen from the medium. Conversely, the overall specific activity of catalase in the cytoplasm did not change appreciably. This indicates that nitrogen depletion induces greater secretion of protein. The vector pCatBex also directs the expression and secretion of CatB in Magnaporthe grisea and may be a useful vector for the expression of genes in other filamentous fungi.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Aspergillus nidulans / genetics*
  • Base Sequence
  • Blotting, Southern
  • Catalase / genetics*
  • Catalase / metabolism*
  • Chromosome Mapping
  • Cladosporium / genetics*
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Fungal Proteins / genetics*
  • Genetic Vectors / genetics*
  • Genetic Vectors / metabolism*
  • Molecular Sequence Data
  • Promoter Regions, Genetic / genetics
  • Protein Sorting Signals / genetics
  • Sequence Analysis, DNA
  • Transformation, Bacterial / genetics

Substances

  • DNA Primers
  • Fungal Proteins
  • Protein Sorting Signals
  • Catalase