title: Improving activity of salt-lyophilized enzymes in organic media.
abstract: Lyophilization with salts has been identified as an important method of activating enzymes in organic media. Using salt-activated enzymes to transform molecules tethered to solid surfaces in organic phase requires solubilization of enzymes in the solvents. Methods of improving performance of salt-lyophilized enzymes, further, via chemical modification, and use of surfactants and surfactants to create fine emulsions prior to lyophilization are investigated. The reaction system used is transesterification of N-acetyl phenylalanine ethyl ester with methanol or propanol. Initial rate of formation of amino acid esters by subtilisin Carlsberg (SC) was studied and found to increase two to sevenfold by either chemical modification or addition of surfactants in certain solvents, relative to the salt (only)-lyophilized enzyme. The method to prepare highly dispersed enzymes in a salt-surfactant milieu also improved activity by two to threefold. To test the effect of chemical modification on derivatization of drug molecules, acylation of bergenin was investigated using chemically modified SC.
authors.affiliation: Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831-6226, USA. borolea@ornl.gov
authors.forename: Abhijeet P
authors.initials: AP
authors.lastname: Borole
authors.forename: Brian H
authors.initials: BH
authors.lastname: Davison
mesh_terms: D004789:Enzyme Activation; D004795:Enzyme Stability; D005612:Freeze Drying; D009930:Organic Chemicals / Q000737:chemistry*; D012492:Salts / Q000737:chemistry*; D013379:Substrate Specificity; D013381:Subtilisins / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*
chemical_list: D009930:Organic Chemicals; D012492:Salts; D013501:Surface-Active Agents; D013381:Subtilisins