Recombinant production of an Aspergillus nidulans class I hydrophobin (DewA) in Hypocrea jecorina (Trichoderma reesei) is promoter-dependent |
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Authors: | Monika Schmoll Christian Seibel Caroline Kotlowski Frank Wöllert Genannt Vendt Burghart Liebmann Christian P Kubicek |
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Institution: | (1) Research Area Gene Technology and Applied Biochemistry, Institute of Chemical Engineering, Vienna University of Technology, 1060 Wien, Austria;(2) BASF Fine Chemicals and Biocatalysis Research, BASF SE, GVF/C - A030, 67056 Ludwigshafen, Germany;(3) Global research Crop Protection, BASF SE, APR/FI - LI470, 67117 Limburgerhof, Germany |
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Abstract: | Fungal hydrophobins have potential for several applications because of their abilities to change the hydrophobicity of different
surfaces. Yet because of their tendency for aggregation and attachment to interfacial areas only few production processes
have so far been reported. Towards the development of a heterologous production system, we report here the expression of a
class I hydrophobin DewA of Aspergillus nidulans in Hypocrea jecorina (Trichoderma reesei). Using the H. jecorina hfb2 (class II hydrophobin-encoding) promoter and lactose as a carbon source, only a minor fraction of the DewA remained cell-wall-bound
and the majority of it secreted into the medium with up to 15% of the total secreted protein. N-terminal amino acid sequencing
showed that it was correctly processed. In contrast, no DewA was produced under the cel7A (cellobiohydrolase I) promoter, although its mRNA was abundantly detected in the cells. This lack of secretion is not due
to trapping in the cell wall or to its degradation because of the unfolded protein response. Recombinant DewA could be conveniently
precipitated from the culture filtrate, and its bioactivity proven by its ability to stably bind to hydrophilic and hydrophobic
surfaces (glass and Teflon, respectively). We thus consider H. jecorina as a promising host for further optimization of DewA production. |
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