本文選題：生物制品　切入點：聚羥基脂肪酸　出處：《汕頭大學》2011年碩士論文

【摘要】：內(nèi)毒素是革蘭氏陰性菌細胞壁外壁中的特有結(jié)構(gòu),是其在生長、分裂或死亡過程中釋放到環(huán)境中的有毒物質(zhì)。內(nèi)毒素分子化學性質(zhì)非常穩(wěn)定,并且在水溶液中易聚合成不同分子量的聚合物,使得生物制品中內(nèi)毒素的去除變得非常困難。聚羥基脂肪酸酯(PHA)顆粒結(jié)合蛋白質(zhì)(PhaP)對PHA以及其它疏水性聚酯具有較強的親和力。人脂多糖結(jié)合蛋白(hLBP)是人體血漿內(nèi)自然存在的內(nèi)毒素受體。在本論文研究中,融合蛋白rhLBP PhaP基因在畢赤酵母GS115中分泌表達。收集到的融合蛋白通過PhaP錨定在PHB顆粒表面,用于水和蛋白質(zhì)溶液中內(nèi)毒素的去除研究。牛血清白蛋白(BSA)溶液,卵清蛋白(ovalbumin)溶液,胰凝乳蛋白酶原(basic proteinα-chymotrypsinogen)溶液三種蛋白質(zhì)溶液作為模式被使用,用于研究蛋白質(zhì)溶液中內(nèi)毒素的去除效率和蛋白質(zhì)的回收率,以及離子強度和酸堿度對去除效果的影響。結(jié)果表明:本論文研究制作的親和顆粒(rhLBP- PhaP-PHB顆粒)的內(nèi)毒素去除率可達90%以上,并且內(nèi)毒素的去除率和蛋白質(zhì)的回收率幾乎不受離子強度的影響,但是酸堿度對其有一定的影響。我們的研究結(jié)果表明,基于rhLBP- PhaP-PHB顆粒的內(nèi)毒素去除系統(tǒng),具有準備簡單,無毒和高效的特點,非常適合蛋白質(zhì)純化工藝中內(nèi)毒素的去除。
[Abstract]:Endotoxin is a unique structure in the outer wall of Gram-negative bacteria. It is a toxic substance released into the environment during growth, division or death. The molecular chemical properties of endotoxin are very stable. And it is easy to polymerize into polymers with different molecular weights in aqueous solutions. It is very difficult to remove endotoxin from biological products. PhaP) has strong affinity to PHA and other hydrophobic polyester. Human lipopolysaccharide binding protein hLBPis human plasma. Endogenous endotoxin receptor. The fusion protein rhLBP PhaP was secreted and expressed in Pichia pastoris GS115. The fusion protein was anchored on the surface of PHB particles by PhaP for the study of endotoxin removal in water and protein solution. Three protein solutions of chymotrypsin solution, basic protein 偽 -chymotrypsinogen solution, were used as models to study the removal efficiency of endotoxin in protein solution and the recovery rate of protein. The effect of ionic strength and pH on the removal of endotoxin was also studied. The results showed that the endotoxin removal rate of the affinity particle (rhLBP- PhaP-PHB) was over 90%. The removal rate of endotoxin and the recovery rate of protein are almost independent of ionic strength, but the pH has a certain effect on it. Our results show that the endotoxin removal system based on rhLBP- PhaP-PHB particles has a simple preparation. The characteristics of non-toxic and high efficiency are very suitable for removing endotoxin in protein purification process.
【學位授予單位】：汕頭大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R346

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本文編號：1656986