鲍鱼内脏多糖的提取、纯化及其抗氧化和抑菌活性

Extraction,Purification and Antioxidant and Antibacterial Activity Analysis of Polysaccharides from Abalone Viscera

以皱纹盘鲍(Haliotis discus hannai Ino)内脏为实验材料,采用碱性蛋白酶、中性蛋白酶、酸性蛋白酶、胃蛋白酶、胰蛋白酶和木瓜蛋白酶6种蛋白酶对其进行酶解提取多糖,结果显示碱性蛋白酶酶解所得多糖得率最高,为6.66%,进而由单因素试验和正交试验确定了碱性蛋白酶提取鲍鱼内脏多糖的最佳工艺条件.将得到的鲍鱼内脏粗多糖进行体外抗氧化活性测定,结果显示:100%清除羟自由基、100%清除2,2-联氮基-双-(3-乙基苯并噻唑啉-6-磺酸)二氨盐(ABTS)自由基以及700 nm处吸光度(A_(700))为0.485时代表的还原能力对应的鲍鱼内脏粗多糖质量浓度分别为9.6,7.5,8.2 mg/mL,表明所得粗多糖具有较好的抗氧化活性.采用双酶解法以提高多糖含量,结果显示胃蛋白酶二次酶解所得的多糖含量最高,且多糖样品在3种抗氧化活性模型中均具有很好的效果.采用Sevage法和反复冻融法去除蛋白,过氧化氢脱除色素,进一步纯化得到高纯度的鲍鱼内脏多糖,并采用双层平板法测定其抑菌活性,结果表明其对金黄色葡萄球菌(Staphyloccocus aureus)、鼠伤寒沙门氏菌(Salmonella typhimurium)、大肠杆菌(Escherichia coil)和铜绿假单胞菌(Pseudomonas aeruginosa)都具有较好的抑制作用.

The polysaccharides were obtained using abalone( Haliotis discus hannai Ino) viscera as the material in this study and enzymolyzed by alkaline protease,neutral protease,vernase,papain,pepsin and trypsin.The results of antioxidant experiments showed that the yield of polysaccharides was the highest up to 6. 66% when alkaline protease was used.Then through the single factor experiment and the orthogonal experiment,the optimal condition for alkaline protease extracting polysaccharides from abalone visceral was determined.Results of antioxidant activity showed that the concentration of primary polysaccharides from abalone viscera for 100% removal of hydroxyl free radical was 9. 6 mg/mL,the concentration for 100% removal of ABTS radical was 7. 5 mg/mL,and the concentration of reducing capacity at A_(700)= 0. 485 was 8. 2 mg/mL. The results indicated that the polysaccharides had a good antioxidant activity. Additionally,the polysaccharide content and antioxidant activity were the highest with secondary enzymolyzing by pepsin.Furthermore,we got the high-purity abalone visceral polysaccharides with Sevage method and repeated freezing and thawing method to remove proteins,and with hydrogen peroxide method for depigment.Finally,double plate method was used to measure antibacterial activity of the abalone visceral polysaccharides,which showed that the purified polysaccharides suppressed reproduction of Staphylococcus aureus,Salmonella typhimurium,Escherichia coli and Pseudomonas aeruginosa efficently.