Shesterenko E. Immobilization of pig liver microsomes on polymeric supports

The dissertation is devoted to investigation of carboxyl esterase-containing pig liver microsomal fraction immobilization on polymeric supports of natural and synthetic origin for creation of 3-hydroxy-1,4-benzodiazepin-2-one esters enantioselective hydrolysis biocatalysts. In the frame of the work execution, the process of carboxylesterase isolation from pig liver was improved, which allowed to obtain an enzyme with esterase activity close to activity of commercial preparation (Sigma Chemical Co., USA, activity 130 U/mg). Methods of carboxylesterase as a part of pig liver microsomal fraction immobilization in polyvinyl alcohol, phyllophorine from з Phyllophora nervosa and sodium alginate were developed. The highly active biocatalysts (65-80 % retaining of initial esterase activity), stable at storage and to elevated temperatures action, were obtained. Biotechnologies of 3-hydroxy-1,4-benzodiazepin-2-one esters enantioselective hydrolysis with a help of free and immobilized microsomal fraction preparation were created. As a result of enzymatic hydrolysis the S-enantiomers of substrates, with increased affinity to central benzodiazepine receptors and anticonvulsant activity, were obtained for the first time. The immobilized pig liver microsomal fraction preparations, as it was shown, catalyzed enantioselective hydrolysis of 3-hydroxy-1,4-benzodiazepin-2-one esters with 50 % degree of transformation during 5-12 cycles of usage in batch mode.