title: X-Ray Crystallography as a Tool to Determine Three-Dimensional Structures of Commercial Enzymes Subjected to Treatment in Pressurized Fluids.
abstract: The study of enzyme function often involves a multi-disciplinary approach. Several techniques are documented in the literature towards determining secondary and tertiary structures of enzymes, and X-ray crystallography is the most explored technique for obtaining three-dimensional structures of proteins. Knowledge of three-dimensional structures is essential to understand reaction mechanisms at the atomic level. Additionally, structures can be used to modulate or improve functional activity of enzymes by the production of small molecules that act as substrates/cofactors or by engineering selected mutants with enhanced biological activity. This paper presentes a short overview on how to streamline sample preparation for crystallographic studies of treated enzymes. We additionally revise recent developments on the effects of pressurized fluid treatment on activity and stability of commercial enzymes. Future directions and perspectives on the the role of crystallography as a tool to access the molecular mechanisms underlying enzymatic activity modulation upon treatment in pressurized fluids are also addressed.
authors.affiliation: EQA/UFSC, Department of Chemical and Food Engineering, Federal University of Santa Catarina, C.P. 476, Florianópolis, SC, CEP 88040-900, Brazil.
authors.forename: Mirian Cristina
authors.initials: MC
authors.lastname: Feiten
authors.affiliation: EQA/UFSC, Department of Chemical and Food Engineering, Federal University of Santa Catarina, C.P. 476, Florianópolis, SC, CEP 88040-900, Brazil.
authors.forename: Marco
authors.initials: M
authors.lastname: Di Luccio
authors.affiliation: moloX, GmbH, D-14195, Berlin, Germany.
authors.forename: Karine F
authors.initials: KF
authors.lastname: Santos
authors.affiliation: EQA/UFSC, Department of Chemical and Food Engineering, Federal University of Santa Catarina, C.P. 476, Florianópolis, SC, CEP 88040-900, Brazil.
authors.forename: Débora
authors.initials: D
authors.lastname: de Oliveira
authors.affiliation: EQA/UFSC, Department of Chemical and Food Engineering, Federal University of Santa Catarina, C.P. 476, Florianópolis, SC, CEP 88040-900, Brazil. jose.vladimir@ufsc.br.
authors.forename: J Vladimir
authors.initials: JV
authors.lastname: Oliveira
mesh_terms: D018360:Crystallography, X-Ray / Q000379:methods; D004798:Enzymes / Q000737:chemistry*; D006874:Hydrostatic Pressure; D000072417:Protein Domains
chemical_list: D004798:Enzymes
keywords: Enzyme; Pressurized fluids; Structure; X-ray crystallography