Study On The Enzymatic Preparation Of Anti-fatigue Active Peptide From Cervus Elaphus Linnaeus Blood

In this research, Cervus elaphus linnaeus blood was studied. The neutral proteinase and caroid were chosen to hydrolyze cervus elaphus linnaeus blood protein to prepare anti-fatigue peptide, whose anti-fatigue and anti-oxidation activity were assessed by loaded swimming test of mice. The products resulting from hydrolysate were isolated by Sephadex G-15, and its molecular weight distribution of the products was detected with HPLC. Then the anti-fatigue and anti-oxidation activities of the separations by loaded swimming test of mice were estimated. In addition, the anti-fatigue mechanism of Cervus elaphus linnaeus blood peptide was discussed.For determination of the craft of enzymolysis, we used Neutral proteinase and caroid to hydrolyze cervus elaphus linnaeus blood protein, respectively, detected the hydrolysis degree and the ability to scavenge free radical of the hydrolysate. Hydrolysate from caroid showed adaptive activity, with the Degree of Hydrolysis (DH) value of 19.8%(named peptide 1). The optimum hydrolysis conditions were followed: pH 7.4,52℃, 90min, concentration of substrate was 8% (w/V), ratio of enzyme/substrate was 12677 U/g substrate .Three hydrolysates of different DH, the velvet antler powder and cervus elaphus linnaeus blood were selected to animal (mice) experiment. Mice were administrated orally with samples' concentration of 150mg /kg.d for 15 days. And then loaded swimming tests of the mice were conducted.The results showed that compared with control group,peptide 1 prolonged the swimming time significantly to 250 min, and the consumption of hepatic glycogen increased to by 38.4%, the concentration of lactic acid in muscle decreased to by 44.9%. The T-AOC ability, GSH-pX activity in the peptide 1 group increased and the concentration of MDA decreased significantly. The results revealed that anti-fatigue and anti-oxidation activity of cervus elaphus Linnaeus blood enhanced notably after being hydrolyzed by caroid. The peptide1 displayed higher anti-fatigue and anti-oxidation activity compared with the other groups.The relative molecular mass of peptide 1 was determined by gel HPLC, distributed between 272 and 550. The essential amino acid of hydrolysate was 55.34%; branch amino acid was 21.90%. The hydrolysate peptides 1 were isolated by Sephadex G-15(1.8×60cm). The optimal isolation conditions were determined. We obtained five divorced peak collection. The anti-fatigue and anti-oxidation activity of peak collections was determined. Peak 1 and 2 were finally decided to be of the highest acivity.