ISOLATION AND CHARACTERIZATION OF PIG LIVER CARBOXYLESTERASE AND ITS APPLICATION IN STEREOSELECTIVE HYDROLYSIS OF 1,4-BENZODIAZEPINE-2-ONE DERIVATIVES S. A. Andronati, E. A. Shesterenko, O. V. Sevastyanov, I. I. Romanovska, E. A. Semenishina, V. I. Pavlo

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ISSN 1995-5537

"Biotechnology" journal V. 4, No. 5, 2011
Р. 71-76, Bibliography 18, Ukrainian.
Universal Decimal classification: 577.152.311/547.8/548.73

ISOLATION AND CHARACTERIZATION OF PIG LIVER CARBOXYLESTERASE AND ITS APPLICATION IN STEREOSELECTIVE HYDROLYSIS OF 1,4-BENZODIAZEPINE-2-ONE DERIVATIVES

S. A. Andronati, E. A. Shesterenko, O. V. Sevastyanov, I. I. Romanovska,
E. A. Semenishina, V. I. Pavlovsky, V. E. Osetrov

A. V. Bogatsky’s Physico-Chemical Institute of National Academy of Sciences of Ukraine, Odesa

Since the commercial preparations of pig liver carboxylesterase are expensive, the development of convenient and available methods of carboxylesterase isolation for usage in stereoselective hydrolysis of new 1.4-benzodiazepine-2-one esters is an essential task. An effective and convenient method for isolation of pig liver microsomal carboxylesterase with protein yield 2.24 mg/g tissue and esterase activity of 149 ?mole•min–1•mg–1 protein is described in paper. With native electrophoresis usage, 19 protein fractions were found in carboxyl esterase preparation, 11 of which (73% of the total protein) possesses esterase activity. For the first time, using pig liver microsomal carboxyl esterase, the stereoselective 3-acetoxy-7-bromo-1-methyl-5 phenyl-1,2-dihydro-3H-1,4-benzodiazepine-2-one hydrolysis was accomplished and thus S-enantiomer of substrate was obtained.

Key words: carboxyl esterase, microsomal fraction, pig liver, electrophoresis, di-(p-nitrophenyl)phosphate, enantioselective hydrolysis, 1,4-benzodiazepine-2-ones.