Crown ether activation of cross-linked subtilisin Carlsberg crystals in organic solvents

The activity of cross-linked subtilisin Carlsberg crystals in the catalysis of peptide bond formation can be significantly enhanced by pretreatment of the enzyme crystals with crown ethers. Soaking of the enzyme crystals in a solution of crown ether in acetonitrile followed by evaporation of the solvent results in an up to 13 times enhanced enzymatic activity. The effects of crown ether treatment under various conditions gives support for the hypothesis that removal of bound water molecules from the active site during the drying process is the origin of the observed enzyme activation. \u

Large activation of serine proteases by pretreatment with crown ethers

Pretreatment of serine proteases by lyophilisation in the presence of crown ethers leads to large enhancements of enzyme activity in organic solvents

Activity and enantioselectivity of serine proteases in transesterification reactions in organic media

The activity and enantioselectivity of -chymotrypsin and subtilisin Carlsberg in the transesterification of a series of N-acetyl-alanine and -phenylalanine esters in cyclohexane were investigated at constant water activity, both in the absence and presence of 18-crown-6. Isosteric variation of the leaving ability of the alcoholate group in the substrates by fluoro substitution provided information about the acylation step of the reaction. For less reactive esters, the rate of acylation is determined by expulsion of the leaving group, whereas for activated esters the rate is dictated by a physical step, most likely a relatively slow conformational change of the enzyme. This makes the enantioselectivity of the enzymes strongly dependent of the substrate activity and the composition of the reaction medium. The catalytic effect of 18-crown-6, observed for all reactions. can be attributed to an enhanced enzyme-substrate binding and an increase of the rate of acylation