ИСТИНА

Nowadays non-waste technologies in synthetic chemistry become more and more popular. Such processes are often carried out using different enzymes. Dehydrogenases represent the large group of enzymes, which are widely used in synthesis of chiral compounds and other useful molecules. Such enzymes need NADH or NADPH as a cofactor and due to high cost of reduced coenzymes a cofactor regeneration system is obligate part in such kind of processes. It was shown that formate dehydrogenase (FDH, EC 1.2.1.2.) is one of the of the best enzymes for NAD(P)H regeneration. FDH catalyses the reaction of formate oxidation to carbon dioxide coupled with reduction of NAD(P)+ to NAD(P)H. The main advantages of FDH are the irreversibility of catalyzed reaction, low price of formate ion and wide pH optimum of activity. Our laboratory has the largest collection of formate dehydrogenases from different sources. Many FDH genes from bacteria, yeasts and plants were cloned and enzymes were expressed in active and soluble forms. Mutant formate dehydrogenases from bacterium Pseudomonas sp.101 show the highest thermal stability as well as activity in comparison with other reported formate dehydrogenases. Now we have focused on eukaryotic genes. The recombinant enzymes from soya Glycine max (SoyFDH), Arabidopsis thaliana (AthFDH), moss Psychomitrella patens (PpaFDH) and yeast Ogataea parapolymorpha (OpaFDH) were obtained by genetic engineering methods. It was revealed, that SoyFDH has the best Michaelis constants among all known FDHs, but it's less thermally stable compared to other FDHs. New mutant forms of SoyFDH with excellent catalytic characteristics and high thermal stability were obtained by protein engineering. Other enzymes (AthFDH, PpaFDH and OpaFDH) are comparable in their stability with majority of bacterial enzymes (but not with PseFDH), so all the new obtained FDHs can be successfully used for cofactor regeneration. This work was supported by grant of Russian President MK-2304.2014.4 and Russian Foundation for Basic Research (grant RFBR 14-04-01665 А and14-04-01625 А). Protein Science, 2015, v.24, № S1, p.174-175.