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A Study on Major Constituent Proteins of Raw Material of Ejiao and Their Bioactivities

Author: ZhangShaoQuan
Tutor: RaoPingFan
School: Fuzhou University
Course: Biochemistry and Molecular Biology
Keywords: Donkey-hide gelatin Donkey Skin Donkey serum albumin Osteoblasts ACE inhibitory activity
CLC: R282
Type: Master's thesis
Year: 2005
Downloads: 522
Quote: 2
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Abstract


Donkey-hide gelatin as a traditional Chinese medicine in China has more than 2,000 years of application history. Due to the lack of research in the basic problems of the protein composition of the donkey-hide gelatin raw materials of animal skin, donkey-hide gelatin pharmacological mechanism has not been clearly elucidated. To this end, the departure from the donkey-hide gelatin raw materials of animal skin, a major component of protein analysis donkey hide, and on this basis, to explore the relevant mechanism of action of the donkey-hide gelatin. First, total protein extract donkey leather TriPure reagent method and SDS method, SDS-PAGE and amino acid sequence analysis of the results show that, donkey hide not only contains the type I collagen, but also contains a lot of donkey serum albumin (DkSA ). Capillary chromatography analysis showed that the animal skin the collagen and DkSA in the presence of combination. These results demonstrated for the first time that now DkSA is a key component of donkey hide. Thus, gelatin, in addition to comprising collagen type I protein and hydrolysates thereof, must also contain the DkSA and its hydrolyzate. From the complexity of the structure and function of the breadth of speculation, DkSA in donkey-hide gelatin wide range of pharmacological effects may play an important role. Based in the body DkSA can flow with each other in a dynamic equilibrium, choose a rich source of raw materials and contains a large number of the donkey plasma serum albumin as the separation and purification of DkSA. The serum obtained a donkey from fresh plasma was centrifuged by freezing, centrifugation, dialysis, and 0.22 micron membrane filter sterilization step Source 15q strong anion exchange chromatography purified by HPLC separation, to get three containing DkSA the component, cLC SDS-PAGE, PAGE electrophoresis identification, are a single component. SDS-PAGE electrophoresis the measurement of its molecular weight 66.OkDa. Finally, the separation and purification DkSA heated for 10 hours at 60 ° C water bath for virus inactivation treatment. Donkey-hide gelatin-based osteoporosis therapy, the DkSA promote rat osteoblast-like cells (UMR 106) proliferative activity were studied. The results show that different concentrations DkSA promote UMR cell proliferation are very obvious concentration 0.03mg/m1 the highest proliferation rate was 87.31%. With inhibition of osteoblast proliferation and differentiation of function, donkey-hide gelatin may be derived from the treatment of osteoporosis in type I collagen and DkSA synergistic effect on osteoblast proliferation and differentiation of type I collagen. Donkey-hide gelatin-based vascular relaxant effect, has the potential to lower blood pressure function, DkSA trypsin hydrolysates ACE activity was studied. The results showed that its hydrolysates having a good ACE-inhibiting activity. Bioinformatics software simulation trypsin degradation the polypeptide fragments DkSA speculate the peptide hydrolysates may contain YLYEVAR potential inhibition of ACE activity. In summary, this thesis donkey-hide gelatin study provides a new perspective, in-depth study based on animal skin protein and its associated protein derived bioactive peptides research will help to clarify the pharmacological mechanism of the donkey-hide gelatin, depth development of the motherland traditional medicine to provide a reference.

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