Addition of substrate-binding domains increases substrate-binding capacity and specific activity of a chitinase from Trichoderma harzianum.
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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Departamento de Genética Facultad de Biologia, Universidad de Servilla, Apartado 1095, E-41080 Servilla, Spain.
Margolles-Clark
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Benítez
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Penttilä
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D000595:Amino Acid Sequence; D001665:Binding Sites; D020134:Catalytic Domain; D002480:Cellulase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D043366:Cellulose 1,4-beta-Cellobiosidase; D002686:Chitin / Q000378:metabolism*; D002688:Chitinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D014026:Nicotiana / Q000201:enzymology; D014242:Trichoderma / Q000201:enzymology*
D011993:Recombinant Fusion Proteins; D002686:Chitin; D002688:Chitinases; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase