The Research Of Resveratrol Metabolic Enzyme In Cabernet Sauvignon Grapevine Leaves

Resveratrol,as a kind of natural polyphenol, which plays an important role in plant disease resistance, human healthy care and so forth. With the development of experimental technology and people's further knowledge to explore the biological activity of resveratrol, resveratrol derivatives and their analogues were found in grape and many other plants, which were thought to be the metabolizable products of resveratrol. and it was taken attention to research realms such as plant adversity physiology, plant pathology and plant breeding as well. However, it is still a vacancy for the metabolizability of resveratrol in the phenylalanine metabolism pathway. Therefore, it is needed to know more about the relationship between resveratrol and its derivatives and analogues.In this paper, we took the grapevine leaves of Cabernet Sauvignon as the experimental material ,to establish the methods of high performance liquid chromatography for resveratrol and its derivatives. We wanted to find out the accumulation rules of resveratrol and its important derivatives after treating the leaves with ultraviolet radiation; two enzymes were comfirmed to be the metabolizable enzyme of resveratrol, which were polyphenol oxidase and peroxidase.In addition, the metabolizing mechanism of the metabolizable enzymes of resveratrol were discussed in the research . The main results are given as follows:(1)The resveratrol, piceid,ε-viniferin,δ-viniferin, pterostilene five stilbenes had a significantly accumulated in grape leaves after UV-C irradiation, there are two peaks of resveratrol synthesis in seventy-two hours, the synthesis peak ofε-viniferin,δ-viniferin, pterostilbene three stilbenes delay to resveratrol and appeared in second peak of resveretrol synthesis besides piceid.(2)The study found that main metabolites of resveratrol isε-viniferin andδ-viniferin through polyphenol oxidase and peroxidase,respectively.But there are many other indirect and ultimate metabolites in the two crud enzymes reaction system. the research have proved that resveratrol have two metabolic pathways at least.(3) The enzyme which uses the resveratrol as substract, was preliminary purification through the chromatograghy of ion exchange fiber (Q-Sepharose FF) and hydrophobic affinity chromatography (Butyl-Sepharose FF), the result of activitive electrophoresis showed that there is a polyphenol oxidase ,it is not multipie and molecular weight about 41.5kD, the metabolites isε-viniferin.(4) Although theδ-viniferin had a significantly accumulated in grape leaves after UV-C irradiation, the result of activitive electrophoresis showed that not new peroxidase produced and induced. The result showed that peroxidase involved in oxidation of resveratrol through inhibition of hydrogen peroxide formation we demonstrated ultraviolet radiation , peroxidase and hydrogen peroxide are indispensable in resveratrol metabolize at the same time .The result above ensured that there were two metabolize pathways of resveratrol.it was fill up blank of the resveratol metabolize in the phenylalanine metabolism pathway. These results settle the important foundation for the further reseach on the stress physiology and regulation mechanism of resveratrol and its derivatives.