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Characterization of a hydrophobin of the ascomycete Paecilomyces farinosus

✍ Scribed by Stefan Lunkenbein; Meike Takenberg; Manfred Nimtz; Ralf G. Berger

Publisher
John Wiley and Sons
Year
2011
Tongue
English
Weight
556 KB
Volume
51
Category
Article
ISSN
0233-111X

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✦ Synopsis

Abstract

The entomopathogenic ascomycete Paecilomyces farinosus (alternative name Isaria farinosa) synthesized a hydrophobin, irrespective of being grown in submerged or surface culture. The protein was extracted using trifluoroacetic acid and purified using preparative HPLC and SDS‐PAGE. Partial sequences were obtained using ESI‐MS/MS. The peptides were used as a start to apply a ‘template switching oligo' protocol to elucidate the complete open reading frame of P. farinosus hydrophobin 1 (pfah1). The deduced protein sequence comprised 107 amino acids (10.7 kDa) including a 16 amino acid long hydrophobic signal peptide, showed a calculated pI of 4.56, and was interrupted by one intron. Phylogenetic analyses revealed relationships to hydrophobins of the ascomycetes Magnaporthe grisea and Metarhizium anisopliae. Based on solubility, hydropathy pattern and phylogeny PfaH1 was assigned to the class Ia hydrophobins. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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