Heterofunctional Methacrylate Beads Bearing Octadecyl and Vinyl Sulfone Groups: Tricks to Obtain an Interfacially Activated Lipase from Thermomyces lanuginosus and Covalently Attached to the Support

Lipase from Thermomyces lanuginosus (TLL) has been immobilized on a methacrylate macroporous resin coated with octadecyl groups (Purolite Lifetech®® ECR8806F). This immobilization protocol gave biocatalyst significantly higher stability than that obtained using octyl agarose. To further improve the features, we tried to covalently immobilize enzyme this support. For purpose, support was activated divinyl sulfone. The results showed at least 1/3 of molecules were not immobilized. solve problem, produced an aminated and then it permitted full covalent previously TLL. use different blocking agents as reaction endpoint (using ethylenediamine, Asp, Gly, Cys) greatly altered functional features (activity, specificity, or stability). example, ethylenediamine increased ratio activity versus R- S-methyl mandelate by three-fold factor. Cys most stable biocatalyst, maintaining close 90% under conditions where just adsorbed maintained less 55%. That way, strategy modify obtaining interfacially layer and, later, vinyl sulfone groups.