ИСТИНА

Enzymes are biocatalysts with unique properties. Enzymes are widely used in fine organic synthesis and drug preparation. Main drawbacks of native enzymes are low stability and low specificity to ”unnatural” compounds. In our laboratory we carry out systematic studies of structure-function relationship of practically important enzymes. Using rational design approach we prepared new biocatalysts or enzymes with improved properties. Formate dehydrogenase (FDH) is used for coenzyme regeneration if processes of chiral synthesis catalyzed by different oxidoreductases. We cloned and expressed new FDHs from bacteria, yeast and plants. The enzymes were crystallized and 3D structures were solved. Protein engineering of the FDHs resulted in mutant enzymes with increased activity and improved KM values and thermal stability. Properties of FDHs in Ionic Liquids were studied. D-amino acid oxidase (DAAO) is used medicine diagnostics of neurodegenerative diseases, in fine organic synthesis and for preparation of 7-amino cephalosporanic acid (7-ACA) from Cephalosporin C. The last one is initiative synton for production of semi- synthetic cephalosporins. Mutant DAAOs with improved stability, new substrate specificity spectrum and increased catalytic activity with Cephalosporin C were obtained. Hydrolase of esters of α-amino acids (АЕН) is alternative enzyme to penicillin acylase. We prepared recombinant AEH which could achieve up to 95% yield in synthesis of α-amino acids cephalosporins. This work was supported by Russian Foundation for Basic Research, projects 14-04-00865, 14-04-00865, 14-04-01312, 14-04-01625, 14-04-01665, 15-54-78035, 16-34-01320 and Italian National Research Council, prot. n. 421- 09-03-2015.