Recombinant production of an Aspergillus nidulans class I hydrophobin (DewA) in Hypocrea jecorina (Trichoderma reesei) is promoter-dependent.
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.
Schmoll
Monika
M
Research Area Gene Technology and Applied Biochemistry, Institute of Chemical Engineering, Vienna University of Technology, 1060, Wien, Austria. mschmoll@mail.zserv.tuwien.ac.at
Seibel
Christian
C
Kotlowski
Caroline
C
Wöllert Genannt Vendt
Frank
F
Liebmann
Burghart
B
Kubicek
Christian P
CP
D001233:Aspergillus nidulans / Q000235:genetics*; D002244:Carbon / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D007785:Lactose / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein; D014242:Trichoderma / Q000235:genetics / Q000378:metabolism*
D003470:Culture Media; C095254:DEWA protein, Asperigillus nidulans; D005656:Fungal Proteins; D011994:Recombinant Proteins; D002244:Carbon; D007785:Lactose