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.
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.
Johnson
Hannah
H
Department of Biological Sciences, Imperial College London, Sir Alexander Fleming Building, Imperial College Road, London SW7 2AZ, UK.
Whiteford
James R
JR
Eckert
Sabine E
SE
Spanu
Pietro D
PD
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002374:Catalase / Q000235:genetics* / Q000378:metabolism*; D002874:Chromosome Mapping; D002963:Cladosporium / Q000235:genetics*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D017422:Sequence Analysis, DNA; D014169:Transformation, Bacterial / Q000235:genetics
D017931:DNA Primers; D005656:Fungal Proteins; D021382:Protein Sorting Signals; D002374:Catalase