title: Bacterial expression of an active class Ib chitinase from Castanea sativa cotyledons.
abstract: Ch3, an endochitinase of 32 kDa present in Castanea sativa cotyledons, showed in vitro antifungal properties when assayed against Trichoderma viride. The characterization of a cDNA clone corresponding to this protein indicated that Ch3 is a class Ib endochitinase that is synthesized as a preprotein with a signal sequence preceding the mature polypeptide. Bacterial expression of mature Ch3 fused to the leader peptide of the periplasmic protein ompT resulted in active Ch3 enzyme. A plate assay was adapted for semi-quantitative determination of chitinase activity secreted from cultured bacteria, which should facilitate the identification of mutants with altered capacity to hydrolyse chitin.
authors.affiliation: Departamento de Biotecnología, E.T.S. Ingenieros de Montes, Ciudad Universitaria, Madrid, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Allona
authors.forename: C
authors.initials: C
authors.lastname: Collada
authors.forename: R
authors.initials: R
authors.lastname: Casado
authors.forename: J
authors.initials: J
authors.lastname: Paz-Ares
authors.forename: C
authors.initials: C
authors.lastname: Aragoncillo
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002688:Chitinases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D018548:Cotyledon / Q000201:enzymology* / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D016415:Sequence Alignment; D014197:Trees; D014242:Trichoderma / Q000187:drug effects / Q000254:growth & development
chemical_list: D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D002688:Chitinases