Preparation And Antioxidant Of Collagen From Deer Bone

In this paper, The bone of Red Deer as raw material for preparing collagen. The characteristics of collagen and their antioxidant activity were studied.(1) Fresh Red Deer bone as raw material, using double enzyme to hydrolyze bone for preparing deer bone powder. On the basis of single factor experiment, the optimum technological conditions for hydrolyzing bone by orthogonal experiment optimization was investigated. the optimum condition of alkaline protease were pH 9.0, temperature 50℃,hydrolyzing 3 hours. And at pH 8.0, temperature 37℃, using Trypsin hydrolyze 2 hours.Under this condition, the hydrolyzing degree was 12.72%. The hydrolyzing degree was1.84 times and 1.39 times using alkaline protease and Trypsin alone, respectively.(2) Preparing collagen of deer bone use acid-enzyme process. with 0.5mol/l acetic acid and 6000 U/g of pepsin in 4℃. On the results of the single factor experiment, the optimum process to prepare collagen of deer bone by response surface optimization experiment was discussed. The results are as following, amount of enzyme is 6000u/g,extraction time was 48 h, solid to liquid ratio was 15:1. under this condition, the yield of hydroxyproline in collagen was 40.86%. UV wavelength scanning was used to analysis the absorption peak of collagen of deer bone, and the strongest absorption peak was 234 nm.conforming to the characteristic absorption peak of collagen at 230 nm. Infrared spectrum scanning results showed the collagen of deer bone contain 4 segments of amide, and with highly hydrogen bonding. Gel forming ability was stronger than collagen of cattle and pig. DSC scanning analysis showed the thermal transition temperature of collagen of deer bone is 119.3 ℃. SDS-PAGE electrophoresis showed the collagen of deer bone containing the molecular weight of 95 KDa collagen, and with a molecular weight greater than 130 KDa collagen, beef bone and pig bone and deer bone roughly the same molecular weight range SEM analysis showed the collagen of deer bone has a spindle type, similar to the collagen of mammals. amino acid composition of collagen of deer bone analysis showed they were typical amino acid of collagen.(3) The purification of collagen were using dialysis tube of 5KDa and Sephadex G-25.Three components were got, they are molecular weight greater than 5KDa, 1-5KDa, less than 1KDa. Three different molecular weight samples showed strong OH and DPPH free radical scavenging ability, and significantly higher than that with positive control. With the increase of collagen concentration, free radical scavenging ability of different molecular weight collagen samples increased gradually. the smallest molecular weight has strongest scavenging ability. The collagen of deer bone has certain effect on scavenging superoxideanion. with the increase of collagen concentration, superoxide anion scavenging rate increased gradually, but compared to VC, superoxide anion scavenging effect is not obvious.