title: Properties of trypsin and of acid phosphatase immobilized in sol-gel glass matrices.
abstract: Trypsin and acid phosphatase-containing silica sol-gel glasses were obtained by mixing a solution of an enzyme with polyethylene glycol (PEG) 6000 and tetramethoxy orthosilicate at room temperature, followed by gelation and drying. Activity of the immobilized trypsin toward small substrates, such as N-benzoyl-L-arginine-4-nitroanilide at its Km, for the best preparations equaled that of the soluble enzyme. Polylysine (M(r) less than or equal to 13,000) and aprotinin (M(r) = 6,500) inhibited this activity. Larger polylysines as well as soybean trypsin inhibitor (M(r) = 20,100) were ineffective. The sol-gel-entrapped trypsin activity was stable when sol-gel glasses were incubated at ambient temperature (pH 7.5) for several months. In comparison, trypsin, immobilized in sol-gel glass by surface adsorption and incubated under the same conditions overnight, was completely autodigested. The firm interaction between the protein molecules and the silica matrix stabilized the enzymes. Thus, the half-life of sol-gel-entrapped acid phosphatase at 70 degrees C (pH 8.0) was two orders of magnitude larger than that of the enzyme in solution. Transparent, mechanically and chemically stable bioactive sol-gel glasses may be used for the development of robust on-line biochemical photodetection sensors and for the purposes of chemical catalysis.
authors.affiliation: A. Silberman Institute of Life Sciences, Hebrew University, Jerusalem, Israel.
authors.forename: S
authors.initials: S
authors.lastname: Shtelzer
authors.forename: S
authors.initials: S
authors.lastname: Rappoport
authors.forename: D
authors.initials: D
authors.lastname: Avnir
authors.forename: M
authors.initials: M
authors.lastname: Ottolenghi
authors.forename: S
authors.initials: S
authors.lastname: Braun
mesh_terms: D000135:Acid Phosphatase; D001709:Biotechnology; D004800:Enzymes, Immobilized; D005782:Gels; D005898:Glass; D014357:Trypsin
chemical_list: D004800:Enzymes, Immobilized; D005782:Gels; D000135:Acid Phosphatase; D014357:Trypsin