牦牛硫酸软骨素分离纯化及其生物活性作用初探

发布时间：2018-01-30 04:41

  本文关键词： 牦牛软骨 提取 纯化 响应面分析 生物活性　出处：《西北民族大学》2017年硕士论文　论文类型：学位论文

【摘要】：目前国内外主要从牛、猪及鲨鱼等动物软骨组织中提取硫酸软骨素,且提取工艺粗糙,产品纯度较低,质量较差。本文以天祝牦牛软骨为原料,优化软骨中硫酸软骨素的提取、纯化等工艺流程,并研究硫酸软骨素的免疫增强效果、降血脂功能和抗凝血作用,为提高牦牛副产品的深加工和利用价值进行了有益的探索。主要试验及结论如下:1、牦牛硫酸软骨素的提取方法优化。以硫酸软骨素纯度为考察指标,对稀碱-酶解法、碱盐-酶解法和超声波辅助碱盐-酶解法等三种提取方法进行比较筛选,并通过单因素及响应面分析确定超声波辅助碱盐提取硫酸软骨素的最佳工艺条件为:料液比为1:6.5(g/mL)、碱液浓度为8%、提取时间为2.7h、Na+浓度为2.5mol/L、提取温度为40℃,CS纯度平均值为55.60%。对木瓜蛋白酶酶解工艺进行单因素及响应面分析优化,确定木瓜蛋白酶酶解的最佳工艺为:酶解温度为36.5℃,酶解pH为6.9,加酶量为1:829,酶解时间为1.5h,CS纯度平均值为71.60%。2、牦牛硫酸软骨素的阴离子交换树脂法纯化工艺优化。最佳工艺条件为:树脂与CS粗提物水溶液体积比为1:1,CS粗提物水溶液浓度为25mg/mL,CS粗提物水溶液流速为1.OmLmmin,吸附时间70min,吸附后将CS粗提物水溶液流出,保持树脂与NaCl溶液体积比为1:5,NaCl溶液浓度为2.5mo]/L,NaCl溶液流速为1.0mL/min,解吸时间90min。纯化物含量结果平均为82.77%。然后选择葡聚糖凝胶G75进行再次纯化,得到的硫酸软骨素的纯度为99.30%。3、牦牛硫酸软骨素生物活性作用研究。对牦牛硫酸软骨素进行免疫增强作用的研究,结果表明:高剂量硫酸软骨素对小鼠血清溶血素有正影响,且低、中、高剂量的硫酸软骨素对脾脏指数及胸腺指数都有所提高,对小鼠体液免疫功能有增强作用。对牦牛硫酸软骨素进行降血脂功能的研究,结果表明:对高血脂症小鼠进行灌胃硫酸软骨素后,可降低小鼠血液中的总胆固醇、甘油三酯、低密度脂蛋白指标,同时升高高密度脂蛋白指标。对牦牛软骨中提取的CS进行抗凝血作用的研究。结果显示CS具有一定的抗凝血作用,但其作用远低于肝素。
[Abstract]:At present, chondroitin sulfate is extracted from chondroitin sulfate from cartilage tissue of cattle, pig and shark at home and abroad, and the extraction process is rough, the purity of the product is low and the quality is poor. This article takes the cartilage of Tianzhu yak as raw material. The extraction and purification of chondroitin sulfate from cartilage were optimized, and the immune enhancement, blood lipid lowering and anticoagulant effect of chondroitin sulfate were studied. In order to improve the value of further processing and utilization of yak by-products, the main experiments and conclusions are as follows: 1. The extraction method of chondroitin sulfate from yak was optimized. The purity of chondroitin sulfate was taken as the index. Three extraction methods, namely dilute alkali-enzymatic hydrolysis, alkaline salt-enzyme hydrolysis and ultrasonic assisted alkali-enzymatic hydrolysis, were compared and screened. The optimum conditions of ultrasonic assisted extraction of chondroitin sulfate by single factor and response surface analysis were as follows: the ratio of material to liquid was 1: 6.5g / mL, and the concentration of alkali was 8%. The extraction time was 2.7 h, the concentration of Na was 2.5 mol / L, and the extraction temperature was 40 鈩，

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