Ginseng, Pilose Antler Peptide Purification, And Active Research

Proteins and polypeptides were isolated and purified from ginseng root(Panax ginseng C.A.Mey.) and velvet antler (Cervus nippon Temminck) using acombination of gel filtration chromatography, ionic exchange chromatography andRP-C18 chromatography technology. The chemistry nature and structure of thoseproteins and polypeptides were discussed by the spectrum technologies of HPLC,MALDI-TOF-MS, ESI-MS and ESI-MS/MS. The biological activity of those proteinsand polypeptides had been carried on using the cell bioactivities experimentalmodel in vitro.1. Five proteins and one polypeptide were obtained from velvet antler. Threeproteins and one polypeptide were obtained from ginseng root. The N-terminalsequence of Protein CNTPⅢand the sequence of polypeptide CNT14 were searchedagainst Swiss-Prot and the Genbank protein sequence databases, and showed thatthe protein sequence homology were smaller than 50% compared with the knownprotein sequence homology. So Protein CNTPⅢand polypeptide CNT14 of velvetantler were new compound.2. The molecular weight of these proteins and polypeptides were determinedby MALDI-TOF-MS mass spectrum and SDS-PAGE. The N terminal sequence of proteinCNTPⅢwas measured by Edman method The sequence of 15% amino acids containingthree alanine residues suggested a generalα-helical structure from secondarystructure prediction. The amino acid sequence of the polypeptide CNT14 of velvetantler and the polypeptide GS9 of ginseng were determined using the ESI-MS/MStechnology. Has studied repeatedly freezing and thawing change of protein CNTPⅠ,extrapolate protein CNTPⅡand CNTPⅢis two subunits of protein CNTPⅠ.3 iccording to the amino acid sequence of velvet polypeptide CNT14, by thesolid phase FMOC synthetic method, the insoluble resin was used as the carrier,the synthesis of the polypeptide has carried on from C end—The N end, and usingthe spectrum technologies of HPLC, MALDI-TOF-MS, ESI-MS and ESI-MS/MS. Confirmedthe synthesis of the polypeptide to be consistent with the amino acid sequence of polypeptide CNT14 which separated from the velvet antler.4 The bioactivity of velvet antler experimental study indicated, proteinCNTPⅣhad the inhibitory action to U20S cell growth. Protein CNTPⅢandpolypeptide CNT14 had the extremely remarkable multiplication function to HT22cell, indicates the two have certain prospect in the anti-senile medicine aspect.Simultaneously, polypeptide CNT14 of velvet antler has the multiplicationfunction to the epidermal cell, the costal cartilage cell, the spleen cell,inferred that CNT14 was possibly one kind of cell growth factor.5 The pharmacology bioactivity experiment of ginseng protein indicated,ginseng protein GSPⅡhad the inhibitory action to the golden yellowstaphylococcus, the ginseng polypeptide GS9 had the inhibitory action to theadheres function between tumor cell and the activation blood platelet.6 Regarding bacteriostasis activity of ginseng protein GSPⅡas a parameter,had discussed the temperature and the pH value change to the bacteriostasisactivity of ginseng protein GSPⅡ, those would provide the theory parameter forthe optimized ginseng processing craft.