美洲大蠊蛋白质提取工艺优化与对伤口愈合的影响

发布时间：2019-02-23 16:44

【摘要】：目的:针对美洲大蠊体内所含蛋白质,采用多种不同提取、筛选和优化的方法进行美洲大蠊蛋白质的提取工艺,为创建高效的美洲大蠊蛋白质提取途径提供技术支持;使用牛血清白蛋白为参照,以考马斯亮蓝法测定美洲大蠊样品中蛋白质含量,比较各个条件下所提取的蛋白质的量;采用换药法,观察和记录美洲大蠊蛋白质对受伤小鼠伤口愈合的影响,并为美洲大蠊在食品、药品方面的拓展研究提供可参依据。方法:采用碱提法、盐提法和胰蛋白酶水解法对美洲大蠊中的蛋白质进行提取,对比各个提取方法对提取率的影响,确定美洲大蠊蛋白质最佳提取条件。实验经考马斯亮兰法对实验中美洲大蠊所提取出的蛋白质含量进行鉴定。利用试验分离得到的美洲大蠊蛋白质进行小鼠伤口愈合换药实验。结果:本实验采用三种提取美洲大蠊蛋白质的方法,其中,碱提法在碱浓度为1.5%,提取温度为80℃,提取时间为1 h时,美洲大蠊蛋白质提取率最高;盐提法在盐浓度为1.0%,提取温度为40℃,提取时间为2h时,美洲大蠊蛋白质提取率最高;酶提法在酶占样本比为0.6%,酶解温度为45℃,酶解时间为2h时,美洲大蠊蛋白质提取率最高。综合实验证明,胰蛋白酶水解法对美洲大蠊蛋白质的提取率是58.87%,高于其他方法。最佳的提取工艺条件为A3B2C1,即酶占样本比为0.6%,酶解温度为45℃下,酶解时间为1 h时,美洲大蠊蛋白质提取提取效率最高,方法最优。小鼠伤口愈合实验表明,美洲大蠊提取的蛋白质对于小鼠伤口的愈合有促进作用,而且使用美洲大蠊蛋白质的小鼠伤口愈合时间比正常情况下伤口愈合的时间缩短了近一半。结论:通过对美洲大蠊蛋白质的提取实验表明,不同提取方法的提取率存在各方面差异。使用胰蛋白酶水解法是美洲大蠊蛋白质提取的最好方法,并且实验条件可以控制。胰蛋白酶水解法是一种易于工业化生产的提取工艺方法,提取得到的美洲大蠊蛋白质可有效的促进小鼠伤口的愈合,具有药用产品开发前景,在治疗烫伤、烧伤等方面的作用也是可以值得预期的
[Abstract]:Objective: to study the extraction process of Blattella Americana protein by different extraction, screening and optimization methods in order to provide technical support for the establishment of an efficient protein extraction pathway for Blattella Americana. Using bovine serum albumin (BSA) as reference, the contents of protein in Blattella Americana samples were determined by Coomassie brilliant blue method, and the amount of protein extracted under different conditions was compared. The effects of Blattella Americana protein on wound healing in injured mice were observed and recorded by the method of dressing change, and reference data were provided for the further study of Blattella Americana in food and medicine. Methods: the protein of Blattella Americana was extracted by alkali extraction, salt extraction and trypsin hydrolysis. The optimum extraction conditions of Blattella Americana protein were determined by comparing the effects of each extraction method on the extraction rate. The contents of protein extracted from the experiment were identified by Coomassie brilliant blue method. The protein isolated from Blattella Americana was used to change medicine for wound healing in mice. Results: three methods of extracting Blattella Americana protein were used in this experiment. The extraction rate of Blattella Americana protein was the highest when alkali concentration was 1.5, extraction temperature was 80 鈩，

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