Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae

Beat Zurbriggen, Michael J. Bailey, Merja E. Penttilä, Kaisa Poutanen, Matti Linko

    Research output: Contribution to journalArticleScientificpeer-review

    57 Citations (Scopus)

    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 languageEnglish
    Pages (from-to)267-278
    Number of pages12
    JournalJournal of Biotechnology
    Volume13
    Issue number4
    DOIs
    Publication statusPublished - 1 Jan 1990
    MoE publication typeA1 Journal article-refereed

    Keywords

    • Downstream processing
    • Pilot production
    • Saccharomyces cerevisiae
    • Trichoderma reesei cellobiohydrolase II

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