title: Thermal denaturation of β-glucosidase B from Paenibacillus polymyxa proceeds through a Lumry-Eyring mechanism.
abstract: β-glucosidase B (BglB), 1,4-β-D: -glucanohydrolase, is an enzyme with various technological applications for which some thermostable mutants have been obtained. Because BglB denatures irreversibly with heating, the stabilities of these mutants are assessed kinetically. It, therefore, becomes relevant to determine whether the measured rate constants reflect one or several elementary kinetic steps. We have analyzed the kinetics of heat denaturation of BglB from Paenibacillus polymyxa under various conditions by following the loss of secondary structure and enzymatic activity. The denaturation is accompanied by aggregation and an initial reversible step at low temperatures. At T ≥ T ( m ), the process follows a two-state irreversible mechanism for which the kinetics does not depend on the enzyme concentration. This behavior can be explained by a Lumry-Eyring model in which the difference between the rates of the irreversible and the renaturation steps increases with temperature. Accordingly, at high scan rates (≥1 °C min(-1)) or temperatures (T ≥ T ( m )), the measurable activation energy involves only the elementary step of denaturation.
authors.affiliation: Departamento de Química, Universidad Autónoma Metropolitana- Iztapalapa, Mexico.
authors.forename: Menandro
authors.initials: M
authors.lastname: Camarillo-Cadena
authors.forename: Georgina
authors.initials: G
authors.lastname: Garza-Ramos
authors.forename: Mariana
authors.initials: M
authors.lastname: Peimbert
authors.forename: Gerardo
authors.initials: G
authors.lastname: Pérez-Hernández
authors.forename: Rafael A
authors.initials: RA
authors.lastname: Zubillaga
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D004795:Enzyme Stability; D044643:Glucan 1,4-beta-Glucosidase / Q000737:chemistry* / Q000378:metabolism; D006358:Hot Temperature; D007700:Kinetics; D056507:Paenibacillus / Q000201:enzymology*; D011489:Protein Denaturation; D020673:Protein Renaturation
chemical_list: D001426:Bacterial Proteins; D044643:Glucan 1,4-beta-Glucosidase