Enzyme activation in organic solvents: co-lyophilization of subtilisin Carlsberg with methyl-beta-cyclodextrin renders an enzyme catalyst more active than the cross-linked enzyme crystals.
In this study we explored the efficiency of the additive methyl-beta-cyclodextrin (M beta CD) to enhance the activity and enantioselectivity of the serine protease subtilisin Carlsberg in organic solvents. These two parameters, measured for different transesterification reactions and in several solvents, are compared with results obtained by using two additional preparations of the same enzyme: lyophilized powder and cross-linked enzyme crystals (CLEC). The results suggest that co-lyophilization of subtilisin with M beta CD preserves the enzyme's active site tertiary structure rendering a highly active and enantioselective catalyst.
Montañez-Clemente
Ileana
I
Department of Chemistry, University of Puerto Rico, College at Humacao, Humacao, Puerto Rico 00791.
Alvira
Edgardo
E
Macias
Minedys
M
Ferrer
Amaris
A
Fonceca
Maricely
M
Rodriguez
Jessica
J
Gonzalez
Anicele
A
Barletta
Gabriel
G
D002384:Catalysis; D003198:Computer Simulation; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism; D003460:Crystallization; D003505:Cyclodextrins / Q000378:metabolism* / Q000494:pharmacology; D004146:Dioxanes / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004951:Esterification; D005612:Freeze Drying / Q000379:methods; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D008958:Models, Molecular; D008968:Molecular Conformation; D012680:Sensitivity and Specificity; D012997:Solvents / Q000494:pharmacology; D013379:Substrate Specificity; D013381:Subtilisins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D047392:beta-Cyclodextrins
D003432:Cross-Linking Reagents; D003505:Cyclodextrins; D004146:Dioxanes; D012997:Solvents; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D013381:Subtilisins; C025223:1,4-dioxane