title: Inactivation of yeast inorganic pyrophosphatase by organic solvents.
abstract: A number of application for enzymes in organic solvents have been developed in chemical processing, food related conversions and analyses. The only unsolved problem related to nonaqueous enzymology is the notion that enzymes in organic solvent are mostly far less active than in water. Therefore, studies concerning the mechanisms by which enzymes are inactivated by organic solvents would reveal a clear understanding of the structure-function relationship of this phenomenon. Here we analyzed the effects of a series of alcohols (methanol, ethanol, 1-propanol and 2-propanol) and acetone on the activity of yeast inorganic pyrophosphatase. We observed that solvents inactivated the enzyme in a dose-dependent manner. This inactivation is also dependent on the hydrophobicity of the solvent, where the most hydrophobic solvent is also the most effective one. The I50 for inactivation by n-alcohols are 5.9 +/- 4, 2.7 +/- 1 and 2.5 +/- 1 M for methanol, ethanol and 1-propanol, respectively. Inactivation was less effective at 37 degrees C than at 5 degrees C, when the I50 for inactivation by methanol, ethanol and 1-propanol are 4.5 +/- 2, 2.1 +/- 2 and 1.7 +/- 1 M, respectively. Our proposal is that solvent binds to the enzyme structure promoting the inactivation by stabilizing an unfolded structure, and that this binding is through the hydrophobic regions of either the protein or the solvent.
authors.affiliation: Laboratório de Enzimologia e Controle do Metabolismo, Departamento de Fármacos, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, 21941-590 Rio de Janeiro, RJ, Brazil.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Grazinoli-Garrido
authors.forename: Mauro
authors.initials: M
authors.lastname: Sola-Penna
mesh_terms: D000096:Acetone / Q000494:pharmacology*; D000438:Alcohols / Q000494:pharmacology*; D002384:Catalysis / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D043564:Inorganic Pyrophosphatase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012997:Solvents / Q000494:pharmacology*; D015003:Yeasts / Q000201:enzymology*
chemical_list: D000438:Alcohols; D004791:Enzyme Inhibitors; D012997:Solvents; D000096:Acetone; D043564:Inorganic Pyrophosphatase