Generation of soluble and active subtilisin and alpha-chymotrypsin in organic solvents via hydrophobic ion pairing.
With very low concentrations of anionic detergents, such as sodium dodecyl sulfate (SDS) and Aerosol OT (AOT), it is possible to solubilize proteases in organic solvents, while retaining enzymatic activity. For example, the SDS-subtilisin BPN' complex catalyzes transesterification of Ac-Phe-OMe in ethanol with a kcat/Km of 36 M-1 s-1 for mutant M1 and 39 M-1 s-1 for the wild type. By comparison, M1 suspended in ethanol is approximately 1000-fold less active, with a kcat/Km of 0.03 M-1 s-1. Similarly, AOT complexes of alpha-chymotrypsin were found to be approximately 1000 times more active (kcat/Km = 100-350 M-1 s-1) than the suspended enzyme.
Meyer
J D
JD
Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Health Sciences Center, Denver, USA.
Kendrick
B S
BS
Matsuura
J E
JE
Ruth
J A
JA
Bryan
P N
PN
Manning
M C
MC
D001665:Binding Sites; D002918:Chymotrypsin / Q000737:chemistry* / Q000378:metabolism*; D002942:Circular Dichroism; D004143:Dioctyl Sulfosuccinic Acid; D004795:Enzyme Stability; D004951:Esterification; D007700:Kinetics; D009154:Mutation; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D012997:Solvents; D013381:Subtilisins / Q000737:chemistry* / Q000378:metabolism*; D013501:Surface-Active Agents
D012997:Solvents; D013501:Surface-Active Agents; D004143:Dioctyl Sulfosuccinic Acid; C103144:acetylphenylalanine methyl ester; D012967:Sodium Dodecyl Sulfate; D010649:Phenylalanine; D013381:Subtilisins; D002918:Chymotrypsin