title: Atomic-Level Quality Assessment of Enzymes Encapsulated in Bioinspired Silica.
abstract: Among protein immobilization strategies, encapsulation in bioinspired silica is increasingly popular. Encapsulation offers high yields and the solid support is created through a protein-catalyzed polycondensation reaction that occurs under mild conditions. An integrated strategy is reported for the characterization of both the protein and bioinspired silica scaffold generated by the encapsulation of enzymes with an external silica-forming promoter or with the promoter expressed as a fusion to the enzyme. This strategy is applied to the catalytic domain of matrix metalloproteinase 12. Analysis reveals that the structure of the protein encapsulated by either method is not significantly altered with respect to the native form. The structural features of silica obtained by either strategy are also similar, but differ from those obtained by other approaches. In case of the covalently linked R5-enzyme construct, immobilization yields are higher. Encapsulation through a fusion protein, therefore, appears to be the method of choice.
authors.affiliation: Magnetic Resonance Center (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, FI (Italy), Fax: (+39) 0554574923.|Department of Chemistry "Ugo Schiff ", University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino, FI (Italy).|Present address: Giotto Biotech S.R.L., Via Madonna del Piano 6, 50019 Sesto Fiorentino, FI (Italy).
authors.forename: Tommaso
authors.initials: T
authors.lastname: Martelli
authors.affiliation: Magnetic Resonance Center (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, FI (Italy), Fax: (+39) 0554574923.|Department of Chemistry "Ugo Schiff ", University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino, FI (Italy).
authors.forename: Enrico
authors.initials: E
authors.lastname: Ravera
authors.affiliation: Magnetic Resonance Center (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, FI (Italy), Fax: (+39) 0554574923.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Louka
authors.affiliation: Giotto Biotech S.R.L., Via Madonna del Piano 6, 50019 Sesto Fiorentino, FI (Italy).|Present address: Magnetic Resonance Center (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, FI (Italy).|Department of Chemistry "Ugo Schiff", University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino, FI (Italy).
authors.forename: Linda
authors.initials: L
authors.lastname: Cerofolini
authors.affiliation: Institute of Biological Chemistry, Department of Chemistry, University of Vienna, Waehringer Str. 38, 1090 Vienna (Austria).
authors.forename: Manuel
authors.initials: M
authors.lastname: Hafner
authors.affiliation: Magnetic Resonance Center (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, FI (Italy), Fax: (+39) 0554574923.|Department of Chemistry "Ugo Schiff ", University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino, FI (Italy).
authors.forename: Marco
authors.initials: M
authors.lastname: Fragai
authors.affiliation: Institute of Biological Chemistry, Department of Chemistry, University of Vienna, Waehringer Str. 38, 1090 Vienna (Austria).
authors.forename: Christian F W
authors.initials: CF
authors.lastname: Becker
authors.affiliation: Magnetic Resonance Center (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino, FI (Italy), Fax: (+39) 0554574923. claudioluchinat@cerm.unifi.it.|Department of Chemistry "Ugo Schiff ", University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino, FI (Italy). claudioluchinat@cerm.unifi.it.
authors.forename: Claudio
authors.initials: C
authors.lastname: Luchinat
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry*; D002384:Catalysis; D004800:Enzymes, Immobilized / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D045726:Metalloproteases / Q000737:chemistry* / Q000378:metabolism; D017640:Silicates / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D004800:Enzymes, Immobilized; D017640:Silicates; D045726:Metalloproteases
keywords: NMR spectroscopy; biomaterials; green chemistry; proteins; silicates