ИСТИНА

Nowadays green technologies using enzymes have become more and more popular. Many enzymes used in biotech industry require non stable and expensive cofactors. One of such molecules is NAD(P)H. That is why the regeneration step for NAD(P)H regeneration is normally included in the enzymatic process. Formate dehydrogenase (FDH, EC 1.2.1.2.) is commonly used for NAD(P)H regeneration process. The advantages of FDH are practical irreversibility of the reaction, high stability and activity, strict specificity. Besides, the product of the reaction is CO2, which can be easily removed from the reaction mixture. There are three ways of including the regeneration step in biotechnology process – use of enzyme mixtures, whole cell biocatalyst – E.coli cells with several co-expressed genes, and fusion artificial enzymes in which two or more polypeptide chains are combined in one by special sequence which is called linker. In the present work we provide the results on obtaining fusion enzymes, included two types of monooxygenases, two types of FDHs and flexible linker. The molecular modeling, expression results and catalytic properties with different substrates are also provided.