title: Stability and proteolytic activity of papain in reverse micellar and aqueous media: a kinetic and spectroscopic study.
abstract: The stability and proteolytic activity of papain were studied in reverse micellar systems, and in aqueous media. In reverse micelles the maximum activity obtained was 80% of the enzyme activity in aqueous solution. Higher papain stability was found in reverse micellar systems compared with that in aqueous solution with half-lives of 24 and 10 days respectively. Electron spin resonance (ESR) spectroscopy studies of aqueous and reverse micellar systems were performed in an attempt to explain the observed enzyme stability and activity profiles. For this purpose a spin label--TEMPOacetamide--was covalently linked to the Cys-25 residue of the papain active center. ESR spectra of labeled papain indicated that catalytic activity of papain could be related to the conformational rigidity near the reaction center. The lower activities obtained in reverse micelles could be a result of the greater degree of mobility and polarity observed in these systems, which can be attributed to papain unfolding. The greater stability found for papain in reverse micelles could be the result of the limited extent of this denaturing process owing to the organized surfactant molecules around the enzyme.
authors.affiliation: Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, Lisboa, Portugal.
authors.forename: L C
authors.initials: LC
authors.lastname: Vicente
authors.forename: R
authors.initials: R
authors.lastname: Aires-Barros
authors.forename: J M
authors.initials: JM
authors.lastname: Empis
mesh_terms: D001586:Benzoylarginine Nitroanilide / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D004795:Enzyme Stability; D006868:Hydrolysis; D007700:Kinetics; D008823:Micelles; D010206:Papain / Q000737:chemistry / Q000378:metabolism*; D012996:Solutions; D014867:Water
chemical_list: D008823:Micelles; D012996:Solutions; D014867:Water; D001586:Benzoylarginine Nitroanilide; D010206:Papain