On enzymatic activity in organic solvents as a function of enzyme history.
Catalytic activities of alpha-chymotrypsin and subtilisin Carlsberg in various hydrous organic solvents were measured as a function of how the enzyme suspension had been prepared. In one method, lyophilized enzyme was directly suspended in the solvent containing 1% water. In another, the enzyme was precipitated from its aqueous solution by a 100-fold dilution with an anhydrous solvent. In most cases, the reaction rate in a given nonaqueous enzymatic system strongly (up to an order of magnitude) depended on the mode of enzyme preparation. The magnitude of this dependence was markedly affected by the nature of the solvent and enzyme. A mechanistic hypothesis proposed to explain the observed dependencies was verified in additional experiments in which the water contents and enzyme history were further varied.
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Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Klibanov
A M
AM
D002918:Chymotrypsin / Q000737:chemistry / Q000378:metabolism; D004121:Dimethyl Sulfoxide; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D019563:Mechanics; D008956:Models, Chemical; D009930:Organic Chemicals; D011487:Protein Conformation; D012997:Solvents / Q000737:chemistry*; D013381:Subtilisins / Q000737:chemistry / Q000378:metabolism; D014867:Water
D004798:Enzymes; D009930:Organic Chemicals; D012997:Solvents; D014867:Water; D013381:Subtilisins; D002918:Chymotrypsin; D004121:Dimethyl Sulfoxide