A novel fungal expression system: Secretion of active calf chymosin from the filamentous fungus trichoderma reesei

Anu Harkki, Jaana Uusitalo, Michael Bailey, Merja Penttilä, Jonathan K.C. Knowles

    Research output: Contribution to journalArticleScientificpeer-review

    109 Citations (Scopus)

    Abstract

    We have studied the expression of a bovine chymosin cDNA in Trichoderma reesei to test the ability of this novel fungal host vector system to express and secrete heterologous gene products. Four different expression plasmids were constructed to determine the optimum manner to fuse the chymosin cDNA to the promoter and the terminator of the major T. reesei cellulase gene, cellobiohydrolase I (cbh1). All four constructions, when transformed into Trichoderma, determined the secretion of a polypeptide corresponding in size to prochymosin and reacting with antichymosin antiserum. This polypeptide had a molecular weight indistinguishable from chymosin and showed milk clotting activity. In preliminary fermentation studies, one transformant secreted as much as 40 mg/1 of active chymosin.
    Original languageEnglish
    Pages (from-to)596-603
    JournalBio/Technology
    Volume7
    Issue number6
    DOIs
    Publication statusPublished - 1 Jan 1989
    MoE publication typeA1 Journal article-refereed

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