Expression and purification of a functionally active class I fungal hydrophobin from the entomopathogenic fungus Beauveria bassiana in E. coli.

Journal of Industrial Microbiology & Biotechnology
Brett H Kirkland, Nemat O Keyhani

Abstract

Hydrophobins represent a class of unique fungal proteins that have low molecular mass, are cysteine rich, and can self-assemble into two-dimensional arrays at water/air interfaces. These highly surface-active proteins are able to decrease the surface tension of water, thus allowing fungal structures to penetrate hydrophobic-hydrophilic barriers. Due to their unusual biophysical properties, hydrophobins have been suggested for use in a wide range of biotechnological applications. Here we describe a simple method for producing a functionally active class I hydrophobin derived from the entomopathogenic fungus, Beauveria bassiana, in an E. coli host. N-terminal modifications were required for proper expression and purification, and the hydrophobin was expressed as a fusion partner to a cleavable N-terminus chitin-binding domain-intein construct. The protein was purified and reconstituted from E. coli inclusion bodies. Self-assembly of the recombinant hydrophobin was followed kinetically using a thioflavin T fluorescence binding assay, and contact angle measurements of purified recombinant hydrophobin protein (mHyd2) films on a variety of substrata demonstrated its surface modification ability, which remained stable for at least 4 m...Continue Reading

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Citations

Jan 27, 2011·Biotechnology Letters·Yifeng Li
Jul 26, 2015·The Protein Journal·Mohammadreza KhalesiGuy Derdelinckx
Jan 8, 2015·Applied Microbiology and Biotechnology·Han A B Wösten, Karin Scholtmeijer
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Apr 17, 2020·Journal of Colloid and Interface Science·Yingying ChengZefang Wang
Mar 4, 2017·Microbiology Spectrum·Brian Lovett, Raymond J St Leger

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