The Research On Purification Of Trypsin Inhibitor And Preparation, Characteristic Of Zinc Chelating Peptides From Whitmania Pigra

Whitmania pigra is included in the national pharmacopoeia of medicinal animals that contains more than 200 kinds of peptides or proteins in which some have value of drug and biological activity. For example, many protease inhibitors such as hirudin that has a strong inhibitory effect to thrombin. In addition, the whitmania pigra has rich protein besides protease inhibitors. Abundant proteins of whitmania pigra can preparing peptides as ideal protein source to produce metal-peptides chelates in order to avoiding protein waste of whitmania pigra that extracted medicinal ingredients. In this study whitmania pigra as a research object. First of all, in the research use a variety of biological separation technologies to gradually isolated and purified a trypsin inhibitors from Whitmania pigra skin mucu and to make some research on its stability 。 Then use whitmania pigra organization as a protein source to prepared zinc-peptides chelates that have biological activity, and make in-depth research to their preparation process and structure. The main contents and results of this experiment are as follows:1.In a certain order by Sephadex G-50, DEAE anion exchange chromatography,Resource 15 reverse phase chromatography techniques, a protein molecular weight of about 36.6KD isolated and purified from Whitmania pigra skin mucus that named as WTI(Whitmania pigra trypsin inhibitor). Through test found WTI not only has a strong protease inhibitory activity also has good p H and thermal stability.2.In this paper, peptides were prepared form enzymatic hydrolysis of Whitmania pigra protein by composite enzyme(papain and flavourzyme) and then made to react with zinc sulfate heptahydrate under certain conditions to produce zinc-peptides chelates that qualitative analysis by sodium sulfide method. Then its antibacterial activity testing found whitmania pigra peptides-zinc chelates have a strong inhibitory effect to concentration of 105 cfu / ml E. coli and Bacillus subtilis that produced inhibition zone respectively 22 mm and 13 mm,.3.Peptides-zinc chelates and raw peptides have been comparative analyzed by ultraviolet, infrared, fluorescence, X diffraction, electron microscopy shows that whitmania pigra peptides′s structure characteristic have a significant changes after chelated zinc ions. Peptides-zinc chelates having some properties of crystals.