Abstract
Endo-β-glucanase I (EGI) of Trichoderma reesei was produced in laboratory and pilot scale using recombinant strains of "bottom-fermenting" Saccharomyces cerevisiae. The gene egl1 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.
Original language | English |
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Pages (from-to) | 133-146 |
Journal | Journal of Biotechnology |
Volume | 17 |
Issue number | 2 |
DOIs | |
Publication status | Published - 1 Jan 1991 |
MoE publication type | A1 Journal article-refereed |
Keywords
- Downstream processing
- Hydrolysis testing
- Pilot scale production
- Saccharomyces cerevisiae
- Trichoderma reesei endo-β-glucanase I