【摘要】：目的:針對(duì)美洲大蠊體內(nèi)所含蛋白質(zhì),采用多種不同提取、篩選和優(yōu)化的方法進(jìn)行美洲大蠊蛋白質(zhì)的提取工藝,為創(chuàng)建高效的美洲大蠊蛋白質(zhì)提取途徑提供技術(shù)支持;使用牛血清白蛋白為參照,以考馬斯亮藍(lán)法測(cè)定美洲大蠊樣品中蛋白質(zhì)含量,比較各個(gè)條件下所提取的蛋白質(zhì)的量;采用換藥法,觀察和記錄美洲大蠊蛋白質(zhì)對(duì)受傷小鼠傷口愈合的影響,并為美洲大蠊在食品、藥品方面的拓展研究提供可參依據(jù)。方法:采用堿提法、鹽提法和胰蛋白酶水解法對(duì)美洲大蠊中的蛋白質(zhì)進(jìn)行提取,對(duì)比各個(gè)提取方法對(duì)提取率的影響,確定美洲大蠊蛋白質(zhì)最佳提取條件。實(shí)驗(yàn)經(jīng)考馬斯亮蘭法對(duì)實(shí)驗(yàn)中美洲大蠊所提取出的蛋白質(zhì)含量進(jìn)行鑒定。利用試驗(yàn)分離得到的美洲大蠊蛋白質(zhì)進(jìn)行小鼠傷口愈合換藥實(shí)驗(yàn)。結(jié)果:本實(shí)驗(yàn)采用三種提取美洲大蠊蛋白質(zhì)的方法,其中,堿提法在堿濃度為1.5%,提取溫度為80℃,提取時(shí)間為1 h時(shí),美洲大蠊蛋白質(zhì)提取率最高;鹽提法在鹽濃度為1.0%,提取溫度為40℃,提取時(shí)間為2h時(shí),美洲大蠊蛋白質(zhì)提取率最高;酶提法在酶占樣本比為0.6%,酶解溫度為45℃,酶解時(shí)間為2h時(shí),美洲大蠊蛋白質(zhì)提取率最高。綜合實(shí)驗(yàn)證明,胰蛋白酶水解法對(duì)美洲大蠊蛋白質(zhì)的提取率是58.87%,高于其他方法。最佳的提取工藝條件為A3B2C1,即酶占樣本比為0.6%,酶解溫度為45℃下,酶解時(shí)間為1 h時(shí),美洲大蠊蛋白質(zhì)提取提取效率最高,方法最優(yōu)。小鼠傷口愈合實(shí)驗(yàn)表明,美洲大蠊提取的蛋白質(zhì)對(duì)于小鼠傷口的愈合有促進(jìn)作用,而且使用美洲大蠊蛋白質(zhì)的小鼠傷口愈合時(shí)間比正常情況下傷口愈合的時(shí)間縮短了近一半。結(jié)論:通過(guò)對(duì)美洲大蠊蛋白質(zhì)的提取實(shí)驗(yàn)表明,不同提取方法的提取率存在各方面差異。使用胰蛋白酶水解法是美洲大蠊蛋白質(zhì)提取的最好方法,并且實(shí)驗(yàn)條件可以控制。胰蛋白酶水解法是一種易于工業(yè)化生產(chǎn)的提取工藝方法,提取得到的美洲大蠊蛋白質(zhì)可有效的促進(jìn)小鼠傷口的愈合,具有藥用產(chǎn)品開發(fā)前景,在治療燙傷、燒傷等方面的作用也是可以值得預(yù)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，

本文編號(hào)：2429019