Studies On The Antioxidant Of Collagen Peptides From Hybrid Abalone(Haliotis Discus Hannai Ino♀×Haliotis Iris ♂)

In the background of embolden developing the marine economy in our country, the status of mariculture industry is also becoming increasingly prominent. As an economic product, with the gradual expansion of the scale of abalone farming and some non-standard operating while breeding, there have risen some issues such as recession of germplasm, declining of production efficiency and so on. In a certain extent, hybridization is an effective method to overcome these problems.Collagen as biological macromolecules, whose activity may not only reflected characteristics of hybrid abalone germplasm, but also do good to enhancing the use of value-added of abalones.Collagen is abundant abalone gastropod in biological macromolecules, abalone conducted by hybridization analysis of collagen activity, not only can understand the characteristics of hybrid abalone germplasm, but also help to enhance the use of value-added Bowley.In this study, the Haliotis iris was introduced from New Zealand as the male parent, hybridized with the local Haliotis discus hannai as female parent, the parents with their hybridized abalone all be used as experimental material. The collagen of abalone gastropod was extracted after pre-treatment. This study investigated the physiological and biochemical properties, structure of collagen, the hydrolysis of abalone collagen peptides, and the antioxidant activity of three abalone collagen peptides in vitro and in vivo. The main results and conclusions are as follows:1. Firstly, the nutritional composition of abalones gastropod was analyzed, then the appropriate method and conditions of collagen extraction were explored. We found out that the Acid-Enzymatic extraction has higher extraction efficiency,the extraction rate of this method was near 80%.The following physiological and biochemical assay include SDS-PAGE, amino acid composition and PI. The results suggested that the hybrids and Haliotis discus hannai collagen was constitute by α1 (110KDa),α2 (100KDa), β chain (210KDa) and y chain (220KDa), otherwise there was only three chains named α1,α2 and β chain in Haliotis iris, which has the same molecular size. Besides, the α2 in Haliotis iris is darker for about 2 times than the other two abalones. 1/3 of the composition of the amino acid was glycine, both proline and hydroxyproline made up about 10% for collagen. The PI of abalone collagen was about 5.2.2. A variety of protease were selected to hydrolyze the abalone collagen, and the results suggested that the neutral protease was the most suitable one for the hydrolysate hydrolyze by neutral protease had the highest efficient in removing superoxide anion radical(O2-·).Then the hydrolyze condition was optimized by Design-expert, and obtain a linear regression equation:Y=65.42+0.35× A+1.16 × B+1.27 × C-0.14 × AB-0.82 × AC+1.17 × BC-2.74 × A2-1.58 × B2-3.91 × C2. The optimum hydrolyze condition was found to be:pH 7.0, temperature of 45℃, enzyme dosage 3400U/g, hydrolysis time 4.2h.3. Collagen peptides were prepared after hydrolyzed from abalones, antioxidant activity in vitro was measured and the experiments suggested that the parent abalones and the hybrid abalone collagen peptides had high reducing power and radical scavenging ability. Haliotis iris showed the highest reducing power and superoxide anion radical scavenging ability (IC50=3.579 mg/mL). A700 of 4mg/ml collagen peptide from Haliotis iris was 1.803, besides the hybrid abalone had the highest hydroxyl radical scavenging capacity (IC50=1.150mg/mL).4. D-galactose induced aging mice serum and liver MDA, T-SOD and GSH-Px content for testing, comparing to explore hybrid abalone gastropod collagen peptides in antioxidant strength of mice.T-SOD and GSH-Px activity in liver and serum of negative control group was significantly decreased compared with the control group, while the MDA content increased significantly. After the mice were feed with the abalone collagen peptides, SOD and GSH-Px antioxidant enzymes vitality increased, and the degree of MDA accumulation decreased, besides, the weight of the thymus increased to normal level. What should be noted is that collagen peptide of hybrid abalone had significant correction effect(P<0.01) on T-SOD in liver, and GSH-Px, MDA content in liver and serum of aging mice.All of these results above show that there was heterosis in free radical scaving and antioxcidant in vivo of hybrid abalone collagen peptides, which may provide a scientific basis and theoretical foundation for the research and development of hybrid abalone functional applications.