Characterization of a hydrophobin of the ascomycete Paecilomyces farinosus

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.

PfaH2: A novel hydrophobin from the ascomycete Paecilomyces farinosus

“…Owing to their amphiphilic nature, hydrophobins can be immobilized on different kinds of surfaces, including clean glass, Teflon, and polystyrene , and are capable of altering the surface properties. The ability of PfaH1 and PfaH2 to change the wetting properties was analyzed with water CA measurements.…”

“…Further isolation and purification of hydrophobins using preparative high‐performance liquid chromatography (HPLC) followed the method described by Ref. . The HPLC system consisted of a Jasco PU‐980 HPLC pump (Jasco, Groß‐Umstadt, Germany), a Jasco LG 980–02 ternary gradient unit, a two line degasser (Techlab, Braunschweig, Germany), and a Jasco MD‐910 multiwavelength detector, all of which were controlled from a Jasco LC‐NetII/ADC box.…”

Growth of the fungus Paecilomyces lilacinus with n-hexadecane in submerged and solid-state cultures and recovery of hydrophobin proteins

“…The PLHYD proteins from the mycelium, produced in both SmC and SSC, were extracted according to a modified version of the procedure described by Lunkenbein et al [20]. Initially, the residual n-hexadecane from SmC and SSC was eliminated with one hexane extraction (1:10).…”