Pilot scale production of a Trichoderma reesei endo-beta-glucanase by brewer's yeast.
Endo-beta-glucanase I (EGI) of Trichoderma reesei was produced in laboratory and pilot scale using recombinant strains of "bottom-fermenting" Saccharomyces cerevisiae. The gene eg/1 was integrated in the chromosome or an expression cassette was inserted on a multicopy plasmid. Expression levels were compared in a laboratory scale bioreactor. The best EGI-producing strain was cultivated in pilot scale. Adsorbent treatment was used to remove endogenous yeast proteins and other impurities from the culture filtrate during concentration. Effective pilot scale one-step purification of the EGI protein was obtained using DEAE-Sepharose, on which EGI was weakly bound. The purified enzyme reacted with antibodies prepared against T. reesei EGI and catalyzed the hydrolysis of both insoluble and soluble substrates.
VTT, Biotechnical Laboratory, Espoo, Finland.
B D
BD
Zurbriggen
M E
ME
Penttilä
L
L
Viikari
M J
MJ
Bailey
D005285:Fermentation; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D005822:Genetic Vectors; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013379:Substrate Specificity; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics
D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D004693:Glucan Endo-1,3-beta-D-Glucosidase