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

doi:10.1111/j.1574-6968.2001.tb10619.x

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

University of Seville

Research output: Contribution to journal › Article › Scientific › peer-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 language	English
Pages (from-to)	57-63
Number of pages	7
Journal	FEMS Microbiology Letters
Volume	198
Issue number	1
DOIs	https://doi.org/10.1111/j.1574-6968.2001.tb10619.x
Publication status	Published - 20 Apr 2001
MoE publication type	A1 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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10.1111/j.1574-6968.2001.tb10619.x

Cite this

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

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.",

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AU - Benítez, Tahía

AU - Penttilä, Merja

PY - 2001/4/20

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N2 - 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.

AB - 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.

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KW - Chitin-binding domain

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KW - Enzyme engineering

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

Abstract

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Keywords

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