title: Structural and catalytic properties of enzymes in reverse micelles.
abstract: Structural and catalytic properties of two enzymes--alpha-chymotrypsin and horse liver alcohol dehydrogenase (LADH)--are studied in bis(2-ethylhexyl) sodium sulfosuccinate (AOT)-isooctane reverse-micelle solutions. Circular dichroism (CD) and electron paramagnetic resonance spectroscopy (EPR) studies show little change in alpha-chymotrypsin structure upon incorporation into reverse micelles. These structural properties explain, in part, the observed activity of these two enzymes in reverse micelles. alpha-Chymotrypsin retains activity in reverse micelles and, in some cases, displays enhanced activity. A sixfold increase in the turnover number was observed in w0 = 10 reverse micelles. LADH has low activity in reverse micelles compared to that in aqueous solution. At w0 = 70, the turnover number of LADH is 18% of the aqueous value. Active-site titrations show a decrease in active enzyme concentration for both enzymes upon incorporation into reverse micelles. Little change in the structure of both LADH and alpha-chymotrypsin is observed with change of water content in the reverse-micelle system.
authors.affiliation: Department of Chemical Engineering, University of California, Berkeley 94720.
authors.forename: A L
authors.initials: AL
authors.lastname: Creagh
authors.forename: J M
authors.initials: JM
authors.lastname: Prausnitz
authors.forename: H W
authors.initials: HW
authors.lastname: Blanch
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002918:Chymotrypsin / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D004143:Dioctyl Sulfosuccinic Acid; D004578:Electron Spin Resonance Spectroscopy; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008823:Micelles; D009795:Octanes; D013501:Surface-Active Agents
chemical_list: D004800:Enzymes, Immobilized; D008823:Micelles; D009795:Octanes; D013501:Surface-Active Agents; D004143:Dioctyl Sulfosuccinic Acid; D000426:Alcohol Dehydrogenase; D002918:Chymotrypsin; C045798:2,2,4-trimethylpentane