Abstract
Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars.
| Original language | English |
|---|---|
| Pages (from-to) | 267-278 |
| Number of pages | 12 |
| Journal | Journal of Biotechnology |
| Volume | 13 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 1 Jan 1990 |
| MoE publication type | A1 Journal article-refereed |
Funding
This work was supporteidn part by Firmenichg rantn o. 83.678.0.8o8f the Swiss National Fund (BZ). The technicaal ssistancoef Tuomo Kuuselai n the pilot hall and of Riitta Isoniemi and Tarja Hakkarainenin the laboratoryis gratefully
Keywords
- Downstream processing
- Pilot production
- Saccharomyces cerevisiae
- Trichoderma reesei cellobiohydrolase II
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