Study On The Bioactive Peptide Of Velvet Antlers And Deer Blood

Deer is a kind of valuable drug dual-use economic animal, and the body is a treasure.The velvet antler, antler blood, deer penis and deer blood have a high drug-use value, but alittle economic value as raw materials. They would obtain dozens of times or even a hundredtimes value, if these raw materials were deep processed.In this paper, antler protein was prepared by enzymatic hydrolysis to obtain antioxidantpeptide. Modern processing technology was also applied to study the extraction andseparation active component in deer blood. The deer industry, the health care industry andrelated medicine industry in our country would receive more opportunities and space fordevelopment when the deep processing technology of antler and deer blood was improved.The main research works include:1. The enzymatic hydrolysis ways were used in preparing the antler active peptides.The antler active peptides were prepared by enzyme hydrolysis and the degree ofhydrolysis and DPPH radical scavenging activity were studied, the optimum technologicalconditions were obtained, and the impact of various factors and their interactions on theexperimental index were analyzed. The hydrolysates had a higher DPPH radical clearancerate when the substrate concentration was between8and14mg/mL, the amount of enzymewas5to8U/mg, the hydrolysis time was55~65min, hydrolysis temperature was about60℃, and the pH was6.2. The membrane separation technology was used in fractionation antler hydrolysateswith different molecular weight cutoff.The membranes with3,5, and10kDa molecular weight cutoff were used to separateantler peptides, and the membrane with0.2kDa was used to concentrate the extract liquid.Three polypeptide segments, peptide I (5-10kDa), peptide II (3-5kDa), and peptide III(0.2-3kDa) were obtained. The several methods of antioxidant activity detection were used.The results showed that: the peptide III (0.2kDa to3kDa) fragment have the strongestability in free radical scavenging. The ability of liposome oxidation inhibition of peptide IIIwas higher than that of Vc when the concentration of peptide exceeded4.0mg/mL; but theability of O2·ˉfree radical scavenging and reducing was weaker than that of Vc.The molecular weight ranges of peptide (peptide I, peptide II, peptide III) were analyzed by Tricine-SDS-PAGE, and the amino acid analyzer was used in analyzing theamino acid composition of the peptide Ⅲ.3. The membrane technology was used in studying separation and purification of deerblood albumin.The technological conditions of membrane technology on separation and purification ofdeer blood albumin were studied. The membrane flux was also studied, including of theoperation pressure, operation time, operation temperature, pH, and total proteinconcentration of the materials. The pretreatment approach of the membrane was used inseparation and purification the deer blood albumin was established. The best operationconditions of the ultra-filtration and the effective membrane cleaning ways were established.The albumin concentrate was frozen dried in the last, the mainly composition of concentratewas protein, accounting for91.5%, the albumin content80.2%, albumin/total protein ratioas high as87.7%. It could be used in developing a natural green albumin healthy food.4. The methodology was studied on ELISA used in detection IGF-1of deer blood andvelvet antlers extract.In this study, the feasibility of ELISA for detection of sika deer plasma and the IGF-1levels in the antler extract liquid were verified methodologically. The minimum detectionlimit was0.4ng/mL, linear range was0.8-200.0ng/mL, intra average coefficient of variationwas6.16%, and between the average coefficients of variation was8.36%. All of these levelswere agreed with the ELISA assay precision. The proposal was that each micro titer plateswere measured and a standard curve was drawn in order to improve the reliability of theELISA assay. The spiked recovery test was launched for determine the recovery of deerplasma and different sample matrix of antler extract. The result showed that all the twosample recovery rates were very high, indicating that the ELISA used in determine IGF-1levels of deer plasma and antler extract had a better accuracy.Deer plasma and antler extract were diluted to different multiples to detect the IGF-1content. The results showed that the IGF-1levels in deer plasma and the reciprocal value ofthe dilution factor were a good linear relationship; the other proteins in deer plasma did notinterfere with the test results. When the antler extract concentration was too high, it wouldinterfere with the cross-reactivity in the ELISA; make the detection of IGF-1measured valuenot accurate. But the interference could be reduced by dilution, therefore the ELISA methodcould be used in detecting IGF-1levels in the deer plasma and antler extract.5. ELISA was used in detecting the IGF-1content of sika deer products. The established ELISA method was applied for detect both antler extract of differentparts and different preservation methods was used in studying deer blood IGF-1stability.The results showed that IGF-1levels in different parts of the antler is top> middle> roots,there are significant differences. The highest position of IGF-1levels was in the top. If deerblood was stored at room temperature for1to3days, the loss rate of IGF-1was relativelyslow. Kept at room temperature for3-7days, the loss rates of IGF-1rose sharply. The IGF-1losses decreased slowly when saved in refrigerated4℃for7days, IGF-1loss rate increasesrapidly after7days. It was recommended that the cold storage time was less than five days,otherwise the loss was large. Kept in-20℃for six months, the loss rate of IGF-1was little.Kept in cryopreservation for a year, the loss rate of IGF-1was a half.