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The Study of Preparing Soluble Eggshell Membrane Protein and Peptide & Antioxidative Activity

Author: ZhouYanHua
Tutor: MaMeiHu
School: Huazhong Agricultural University
Course: Agricultural Products Processing and Storage
Keywords: Eggs Shell membrane Soluble egg shell membrane protein Light Antioxidant peptide
CLC: TQ464.7
Type: Master's thesis
Year: 2010
Downloads: 195
Quote: 0
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Abstract


This subject innovative use of The secondary extract prepared soluble egg shell membrane protein, and studied its physicochemical properties; using various protease digested, studied the enzymatic hydrolysis of egg shell membrane protein preparation of a polypeptide, the best process; using chemiluminescence method for the determination of the antioxidant activity of the egg shell membrane protein hydrolysates; Sephadex G-25 isolated components with high antioxidant activity, and of each component were determined by gel chromatography The average molecular weight. The results are as follows: the secondary extraction method to extract soluble egg shell membrane protein. Optimized by single factor and orthogonal experiment results show that the time to restore the extraction rate of 17.80%. Secondary pepsin extracted egg shell membrane protein extraction rate can be as high as 75.82%. The extraction rate of 93.62% through secondary reactions egg shell membrane protein can basically achieve the purpose of use of the whole film. The determination of the physical and chemical properties of soluble egg shell membrane protein. By UV, the scan results of FT-IR spectroscopy and circular dichroism (CD) scan results show that the the prepared egg shell membrane protein has a more complete protein secondary structure. Amino acid analyzer and determination of its amino acid composition, indicating that the egg shell membrane protein containing all the essential amino acids the human body, is a nutritious, high-quality protein. Also measured the water absorption of the egg shell membrane proteins, Paul aqueous emulsification and foaming properties and other functional properties of the measurement results for the water-absorbent 0.057g / g, emulsifiable 6.92 m 2 / gpro, the results were better than the control gelatin, egg shell membrane protein is a great development potential protein resource. Various enzyme soluble egg shell membrane protein hydrolysis effect comparative study, the results show that: the degree of hydrolysis and nitrogen yield of indicators, select the alkaline protease and papain enzyme hydrolysis has better enzymatic effect The quadratic regression orthogonal rotary draw the following test conclusion: alkaline proteases: papain ratio of 8:2 has joined, the total amount of the enzyme for 16000 U / g, alkaline protease enzymatic time 2 h, the Papain the enzymatic hydrolysis time 1 h, the alkaline proteases pH 9.0, papain, pH 5.5, hydrolysis temperature 50 ° C, the degree of hydrolysis of egg shell membrane protease hydrolyzate prepared under the conditions and nitrogen yield of 46.12 percent, respectively, 85.56%. The trial also measured in four different chemiluminescence system eggshell membrane protein and its enzymatic hydrolyzate of superoxide anion, hydroxyl radical, hydrogen peroxide, and DNA damage protective effect of antioxidant activity. Comprehensive experimental results show that the The alkaline protease enzymatic liquid has strong antioxidant activity. Therefore, this subject will continue to optimize the process conditions of the preparation of alkaline protease hydrolysis egg shell membrane protein antioxidant activity peptide, enzymatic hydrolysis time 4h temperature of 55 ° C, pH value of 9.0, it is clear the O2, OH and DNA Damage half of the protective effect suppression the concentration IC 50 0.157mg/mL 0.215mg/mL and 1.128mg/mL. Its antioxidant activity than the egg shell membrane protein in the enzymatic hydrolysis has been greatly enhanced. Compared with other raw materials enzymatic solution, the egg shell membrane protease hydrolyzate antioxidant activity several times or even hundreds of times. Using gel exclusion chromatography separation, enzymatic hydrolyzate of the alkaline protease to give three main component, SP1, SP2, SP3, and the average molecular weight of 10522.14 Da, 618.86 Da and 300.83 Da, respectively, the proportion of 26.59%, 68.15% and 5.62%, respectively. The antioxidant activity of the respective components were measured by chemiluminescence, the test results show that the second component SP2 having the strongest antioxidant activity. Determination of the SP2 clear O 2 - OH. Capacity and DNA damage half inhibitory concentration (IC 50 , the test results were 0.10, 0.18 and 0.95mg/ml.

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CLC: > Industrial Technology > Chemical Industry > Pharmaceutical chemical industry > Drug production of biological products > Amino acids,peptides,proteins
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