Efficient secretion of two fungal cellobiohydrolases by Saccharomyces cerevisiae.
Two different cellobiohydrolases, CBHI and CBHII, of the filamentous fungus Trichoderma reesei both hydrolyse highly crystalline cellulose. Cellulolytic strains of the yeast Saccharomyces cerevisiae were constructed by transferring cDNAs coding for these enzymes into yeast on an expression plasmid. These cellulolytic yeasts were able to secrete efficiently the large, heterologous proteins to the culture medium. The recombinant cellulases were observed to be heterogeneous in Mr due, at least partly, to variable N-glycosylation. Recombinant CBHII was able to bind to crystalline cellulose, although slightly less efficiently than the native enzyme. Both of the two recombinant cellulases were able to degrade amorphous cellulose. In a fermenter cultivation, around 100 micrograms/ml of CBHII was secreted into the yeast growth medium.
Penttilä
M E
ME
VTT Biotechnical Laboratory, Espoo, Finland.
André
L
L
Lehtovaara
P
P
Bailey
M
M
Teeri
T T
TT
Knowles
J K
JK
D002482:Cellulose / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase; D004247:DNA / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D012636:Secretory Rate; D014242:Trichoderma / Q000235:genetics
D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D002482:Cellulose; D004247:DNA; D006026:Glycoside Hydrolases; D043366:Cellulose 1,4-beta-Cellobiosidase