Process technological effects of deletion and amplification of hydrophobins I and II in transformants of Trichoderma reesei.
Transformants of the Trichoderma reeseistrains QM9414 and Rut-C30 were constructed in which the genes for the two major hydrophobin proteins, hydrophobins I (HFBI) and II (HFBII), were deleted or amplified by molecular biological techniques. Growth parameters and foam production of the transformant strains were compared with the corresponding properties of the parent strains by cultivation in laboratory bioreactors under conditions of catabolite repression (glucose medium) or induction of cellulolytic enzymes and other secondary metabolites (cellulose and lactose media). All the transformed strains exhibited vegetative growth properties similar to those of their parent. The Delta hfb2 (but not the Delta hfb1) transformant showed reduced tendency to foam, whereas both strains overproducing hydrophobins foamed extensively, particularly in the case of HFBII. Enzyme production on cellulose medium was unaltered in the Delta hfb2 transformant VTT D-99676, but both the Delta hfb2 and HFBII-overproducing transformants exhibited somewhat decreased enzyme production properties on lactose medium. Production of HFBI by the multi-copy transformant VTT D-98692 was almost 3-fold that of the parent strain QM9414. Overproduction of HFBII by the transformant VTT D-99745, obtained by transformation with three additional copies of the hfb2 gene under the cbh1 promoter, was over 5-fold compared to production by the parent strain Rut-C30. The Delta hfb2transformant VTT D-99676 produced a greatly increased number of spores on lactose medium compared with the parent strain, whereas the HFBII-overproducing transformant VTT D-99745 produced fewer spores.
Bailey
M J
MJ
VTT Biotechnology, Box 1500, 02044 VTT, Finland. michael.bailey@vtt.fi
Askolin
S
S
Hörhammer
N
N
Tenkanen
M
M
Linder
M
M
Penttilä
M
M
Nakari-Setälä
T
T
D001483:Base Sequence; D002480:Cellulase / Q000096:biosynthesis; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification; D005784:Gene Amplification; D017353:Gene Deletion; D020033:Protein Isoforms / Q000235:genetics* / Q000302:isolation & purification; D013171:Spores, Bacterial; D014170:Transformation, Genetic; D014242:Trichoderma / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000502:physiology
D017931:DNA Primers; D005656:Fungal Proteins; D020033:Protein Isoforms; D002480:Cellulase