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Applied Microbiology and Biotechnology

, Volume 57, Issue 1–2, pp 124–130 | Cite as

Overproduction, purification, and characterization of the Trichoderma reesei hydrophobin HFBI

  •  S. Askolin
  •  T. Nakari-Setälä
  •  M. Tenkanen
Original Paper

Abstract.

Many characteristics of fungal hydrophobins, such as an ability to change hydrophobicity of different surfaces, have potential for several applications. The large-scale processes of production and isolation of these proteins susceptible to aggregation and attachment to interfacial surfaces still needs to be studied. We report for the first time on a method for a gram-scale production and purification of a hydrophobin, HFBI of Trichoderma reesei. A high production level of the class II hydrophobin (0.6 g l–1) was obtained by constructing a T. reesei HFBI-overproducing strain containing three copies of the hfb1 gene. The strain was cultivated on glucose-containing medium, which induces expression of hfb1. HFBI hydrophobin was purified from the cell walls of the fungus because most of the HFBI was cell-bound (80%). Purification was carried out with a simple three-step method involving extraction of the mycelium with 1% SDS at pH 9.0, followed by KCl precipitation to remove SDS, and hydrophobic interaction chromatography. The yield was 1.8 g HFBI from mycelium (419 g dw), derived from 15 l of culture. HFBI was shown to be rather unstable to N-terminal asparagine deamidation and also, to some extent, to non-specific proteases although its thermostability was excellent.

Keywords

Purification Hydrophobic Interaction Asparagine Trichoderma Production Level 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  •  S. Askolin
    • 1
  •  T. Nakari-Setälä
    • 1
  •  M. Tenkanen
    • 1
  1. 1.VTT Biotechnology, PO Box 1500, 02044 VTT, Finland

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