title: Overexpression, purification and characterization of the Trichoderma atroviride endochitinase, Ech42, in Pichia pastoris.
abstract: The endochitinase gene ech42 from Trichoderma atroviride was cloned and expressed in Pichia pastoris using a constitutive expression system. Over 98% of the recombinant protein was secreted into the culture medium as glycoprotein. A high endochitinase concentration, 186 mg/L with a specific enzyme activity of 14,128 Umg(-1) was produced. The optimal enzyme kinetic parameters for the recombinant protein were identical to those reported for the enzyme isolated from T. atroviride. The recombinant endochitinase possesses suitable features for biotechnological applications, such as activity at acidic pH and thermostability.
authors.affiliation: Institute for Scientific and Technological Research in San Luis Potosí, Molecular Biology Division, Camino a la Presa San José 2033, Lomas 4a sección, CP 78216 San Luis Potosí, Mexico.
authors.forename: Ana S
authors.initials: AS
authors.lastname: Pérez-Martínez
authors.forename: Antonio
authors.initials: A
authors.lastname: De León-Rodríguez
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Harris
authors.forename: Alfredo
authors.initials: A
authors.lastname: Herrera-Estrella
authors.forename: Ana P
authors.initials: AP
authors.lastname: Barba de la Rosa
mesh_terms: D002384:Catalysis; D002688:Chitinases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D010843:Pichia / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D002688:Chitinases