title: Immobilization of a recombinant cutinase by entrapment and by covalent binding. Kinetic and stability studies.
abstract: Fusarium solani pisi recombinant cutinase, immobilized by entrapment in calcium alginate and by covalent binding on porous silica, was used to catalyze the hydrolysis of tricaprylin. The influence of relevant parameters on the catalytic activity such as pH, temperature, and the substrate concentration were studied. Cutinase immobilized by entrapment presented a Michaelis-Menten kinetics for tricaprylin concentrations up to 200 mM. At higher concentrations of substrate, inhibition was observed. For covalent binding immobilization, diffusional limitations were observed at low substrate concentrations and substrate inhibition occurred for concentrations higher than 150 mM. The stability of immobilized cutinase was also evaluated. The enzyme immobilized by entrapment showed a high stability, in contrast to the immobilization on porous silica.
authors.affiliation: Laboratório de Engenharia Bioquimica, Instituto Superor Técnico, Lisboa, Portugal.
authors.forename: A P
authors.initials: AP
authors.lastname: Gonçalves
authors.forename: J M
authors.initials: JM
authors.lastname: Cabral
authors.forename: M R
authors.initials: MR
authors.lastname: Aires-Barros
mesh_terms: D002210:Caprylates / Q000008:administration & dosage / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005079:Excipients / Q000378:metabolism; D005670:Fusarium; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011994:Recombinant Proteins / Q000378:metabolism; D012822:Silicon Dioxide; D013696:Temperature; D014280:Triglycerides / Q000008:administration & dosage / Q000378:metabolism
chemical_list: D002210:Caprylates; D004800:Enzymes, Immobilized; D005079:Excipients; D011994:Recombinant Proteins; D014280:Triglycerides; C003637:tricaprylin; D012822:Silicon Dioxide; D002265:Carboxylic Ester Hydrolases; C019952:cutinase