Addition of substrate-binding domains increases substrate-binding capacity and specific activity of a chitinase from Trichoderma harzianum

M. Carmen Limón, Emilio Margolles-Clark, Tahía Benítez, Merja Penttilä*

*Corresponding author for this work

    Research output: Contribution to journalArticleScientificpeer-review

    79 Citations (Scopus)

    Abstract

    Chitinase Chit42 from Trichoderma harzianum CECT 2413 is considered to play an important role in the biocontrol activity of this fungus against plant pathogens. Chit42 lacks a chitin-binding domain (ChBD). We have produced hybrid chitinases with stronger chitin-binding capacity by fusing to Chit42 a ChBD from Nicotiana tabacum ChiA chitinase and the cellulose-binding domain from cellobiohydrolase II of Trichoderma reesei. The chimeric chitinases had similar activities towards soluble substrate but higher hydrolytic activity than the native chitinase on high molecular mass insoluble substrates such as ground chitin or chitin-rich fungal cell walls.

    Original languageEnglish
    Pages (from-to)57-63
    Number of pages7
    JournalFEMS Microbiology Letters
    Volume198
    Issue number1
    DOIs
    Publication statusPublished - 20 Apr 2001
    MoE publication typeA1 Journal article-refereed

    Funding

    We thank Dr. Anu Koivula and Dr. Markus Linder (VTT Biotechnology, Espoo, Finland) for advice on the binding assays. This work was financially supported by the grants from Junta de Andalucı́a PAI CVI 107, Universidad Internacional de Andalucı́a, CICYT BIO94-0289, BIO97-0521, IFD97-0668 and PTR94-0068. M.C.L. was recipient of a short term fellowship from the Spanish Ministry of Education and Culture.

    Keywords

    • Cellulose-binding domain
    • Chitin-binding domain
    • Chitinase
    • Enzyme engineering
    • Trichoderma harzianum

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