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Интеллектуальная Система Тематического Исследования НАукометрических данных |
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Cloning, expression and characterization of new NAD+-dependent formate dehydrogenase from moss Physcomitrella patensтезисы доклада
- Авторы: Dolina I.A., Alekseeva A.A., Savin S.S., Tishkov V.I.
- Сборник: Program and Abstract Book of International Conference "Biocatalysis-2015: Fundamentals and Applications", Moscow region, Avantel Club Istra
- Тезисы
- Год издания: 2015
- Место издания: Innovations and High Technologies MSU Ltd Moscow
- Первая страница: 103
- Последняя страница: 103
- Аннотация: Formate dehydrogenase (EC 1.2.1.2, FDH) catalyzes oxidation of formate ion to carbon diox-ide coupled with the reduction of NAD+ to NADH. FDH was found in bacteria, yeast, microscopic fungi and plant. Formate dehydrogenase is an enzyme of a great interest for fundamental science and as well as for biotechnology. The enzyme is used for reduced cofactor regeneration in processes of fine organic synthesis with NAD(P)+-dependent oxidoreductases. In this laboratory we study FDH from different sources for a long time. Presently special attention is paid to plant formate dehydrogenases. FDH from plants have unique kinetic properties in comparison with those of other organisms. For further research we chose moss P. patens as the source of FDH as it is a model organism for studying the evolution and physiology of plants. The primary structure of the signal peptide of the enzyme from moss is significantly different compared to those of enzymes from other plants. In plants FDH is a stress protein and it also has been shown for the enzyme from the moss P. patens. In this work we cloned FDH gene of moss P. patens (PpaFDH) by the method of reverse tran-scription- PCR. As a result, the genetic construct containing the cDNA of the gene encoding the full-size variant of the enzyme was created. Also the expression of PpaFDH in E. coli cells was done, and as a result we received the enzyme in an active and soluble form with formate dehydrogenase activity. The conditions of FDH purification were chosen and the basic properties of enzyme, such as kinetic parame-ters and thermal stability, were studied. In addition, a comparison of the properties of this enzyme with properties of formate dehydrogenases from other sources was made. The work was supported by Russian Foundation for Basic Research (RFBR) (grant 14-04-01665-a).
- Добавил в систему: Тишков Владимир Иванович