title: Solvent effects on the catalytic activity of subtilisin suspended in organic solvents.
abstract: We studied a model transesterification reaction catalyzed by subtilisin Carlsberg suspended in toluene, n-hexane, diisopropyl ether, and mixtures of these solvents. To account for solvent effects due to differences in water partitioning between the enzyme and the bulk solvents, we measured water sorption isotherms for the enzyme in each solvent. We measured catalytic activity as a function of enzyme hydration and obtained bell-shaped curves with maxima at the same enzyme hydration in all the solvents. However, the activity maxima were different in all the media, being the lowest in toluene. Differences in the partitioning of substrates and product between the bulk solvent phase and the enzyme active site were accounted for but could not explain the lower catalytic activity observed in toluene. The fact that toluene is very similar to one of the substrates suggested the possibility of competitive inhibition by this solvent. We derived a model allowing for differences in solvation of the substrates, by using thermodynamic activities instead of concentrations, as well as for competitive inhibition by toluene. The model fit the experimental data well, confirming that toluene had a direct adverse effect on the catalytic activity of the enzyme.
authors.affiliation: Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, R. da Quinta Grande, 6, Apt. 127, 2780 Oeiras, Portugal.
authors.forename: T C
authors.initials: TC
authors.lastname: de Sampaio
authors.forename: R B
authors.initials: RB
authors.lastname: Melo
authors.forename: T F
authors.initials: TF
authors.lastname: Moura
authors.forename: S
authors.initials: S
authors.lastname: Michel
authors.forename: S
authors.initials: S
authors.lastname: Barreiros