本文選題：木質(zhì)纖維　切入點：接枝改性　出處：《北京化工大學(xué)》2015年碩士論文

【摘要】：膜色譜兼具傳統(tǒng)柱色譜的高選擇性,以及膜分離技術(shù)高效率、低壓降、易放大實施的優(yōu)越性,相比于柱色譜更適合用于大規(guī)模蛋白分離純化。疏水作用膜色譜利用不同蛋白分子疏水性的差異導(dǎo)致與分離膜的作用力不同達(dá)到分離純化目的,分離條件溫和,有利于保持蛋白的活性。本研究以紙纖維為基材,通過接枝改性獲得紙基疏水色譜分離膜,并考察所得分離膜對混合蛋白的分離性能。主要研究工作如下：1)采用紫外光引發(fā),將異丙基丙烯酰胺(NIPAM)及NIPAM與丙烯酸丁酯(BA)共聚物大分子單體(P(NIPAM-BA),平均分子量5276Da, BA含量為5.96%)接枝至濾紙上,得到平均接枝率為4.5%、6.7%、12.2%的紙基溫敏色譜膜。通過紅外光譜測試,證明濾紙上成功接枝了溫敏支化聚合物。2)考察了溫敏膜色譜純化脂肪酶粗酶的適宜條件。選用接枝率12%左右的溫敏膜,在流動相無鹽條件下考察色譜膜的分離性能。膜組件于30℃逐漸升溫至40℃并維持加熱10min后往流動相注入脂肪酶粗酶溶液進(jìn)行高溫吸附,于4℃進(jìn)行低溫淋洗脫附,收集淋洗流出蛋白溶液。SDS-PAGE測試結(jié)果表明,經(jīng)溫敏膜色譜的純化,脂肪酶粗酶中的雜蛋白全部在吸附階段得以分離,脫附淋洗液中含有高純度日標(biāo)蛋白,純化效果良好；橄欖油乳化液水解滴定法測定表明,經(jīng)溫敏色譜膜純化后的脂肪酶酶的比活提高了10.9倍。3)采用絲光處理、環(huán)氧開環(huán)聚合方法制備的聚氧化苯乙烯接枝改性木質(zhì)纖維制備了紙基疏水色譜膜(PSO-g-PF膜),考察了該疏水色譜膜的性能。光學(xué)顯微鏡及SEM對其形態(tài)圖像觀測表明,PSO-g-PF膜的微觀形貌與商品化濾紙及未改性紙漿纖維網(wǎng)類似,具備紙纖維膜所特有的孔形態(tài)結(jié)構(gòu)和高通透性。膜組件分離實驗結(jié)果顯示,在流動相為1.4M硫酸銨鹽溶液時,PSO-g-PF膜對BSA的吸附量達(dá)3.7mg·g-1,為未接枝改性的紙漿纖維膜的1.8倍,表明PSO-g-PF膜適合用作疏水色譜蛋白分離膜,而且重復(fù)使用次數(shù)可達(dá)3-4次。4)研究了PSO-g-PF疏水色譜膜對混合蛋白的分離性能。對于BSA/溶菌酶混合蛋白,吸附階段流動相為0.9M硫酸銨鹽溶液時,可將二者成功拆分；分離黑曲霉脂肪酶／溶菌酶混合蛋白時,吸附階段流動相為1.0M硫酸銨鹽溶液時,可將二者成功拆分；分離黑曲霉脂肪酶/BSA混合蛋白時,吸附階段流動相為0.7M硫酸銨鹽溶液時,可從混合蛋白中去除黑曲霉脂肪酶,得到純BSA；純化脂肪酶粗酶時,吸附階段流動相為1.1M硫酸銨鹽溶液時,能去除雜蛋白,得到高純度的脂肪酶,其比活提高了12.1倍。
[Abstract]:Membrane chromatography has the advantages of high selectivity of traditional column chromatography, high efficiency of membrane separation technology, low pressure drop and easy amplification. The hydrophobic interaction membrane chromatography is more suitable for the separation and purification of large scale proteins than column chromatography. The hydrophobicity of different protein molecules leads to the separation and purification of the membrane with different forces, and the separation conditions are mild. The paper hydrophobic chromatographic separation membrane was obtained by graft modification, and the separation performance of the separation membrane to the mixed protein was investigated. The main research work is as follows: 1) UV light was used to initiate the separation. The copolymer of isopropylacrylamide (NIPAM) and butyl acrylate (NIPAM) was grafted onto the filter paper with an average molecular weight of 5276Daand BA content of 5.96. the paper based thermochromatographic film with an average grafting rate of 4.56.7and 12.2% was obtained. It was proved that the temperature sensitive branched polymer. 2) the suitable conditions for the purification of lipase by thermo-sensitive membrane chromatography were investigated. The temperature sensitive membrane with grafting rate of about 12% was selected. The separation performance of chromatographic membrane was investigated under the condition of mobile phase without salt. The membrane module was heated up to 40 鈩，

本文編號：1655108