title: (Hyper)thermophilic enzymes: production and purification.
abstract: The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our understanding and presented new opportunities for solving one of the most challenging problems in biophysics: how is structural stability and biological function maintained at high temperatures where "normal" proteins undergo dramatic structural changes? In our laboratory we have purified and studied many thermostable and hyperthermostable proteins in an attempt to determine the molecular basis of heat stability. Here, we present methods to express such proteins and enzymes in E. coli and provide a general protocol for overproduction and purification. The ability to produce enzymes that retain their stability and activity at elevated temperatures creates exciting opportunities for a wide range of biocatalytic applications.
authors.affiliation: Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands.
authors.forename: Pierpaolo
authors.initials: P
authors.lastname: Falcicchio
authors.forename: Mark
authors.initials: M
authors.lastname: Levisson
authors.forename: Servé W M
authors.initials: SW
authors.lastname: Kengen
authors.forename: Sotirios
authors.initials: S
authors.lastname: Koutsopoulos
mesh_terms: D055162:Biocatalysis; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004798:Enzymes / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D004798:Enzymes