本文選題：殼聚糖　切入點：硅膠　出處：《青島科技大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 本論文分三部分,第一部分,基于大孔生物材料的染料親和色譜,發(fā)展了一種新型的蛋白質(zhì)色譜復(fù)性新技術(shù);第二部分,基于溶膠-凝膠法,通過氨水溶液簡單處理制備了球形硅膠支載的固定化金屬親和吸附劑;第三部分,基于溶膠-凝膠法,通過氫氧化鈉溶液簡單處理制備了球形硅膠支載的生物吸附劑。 1、基于大孔生物材料的染料親和色譜(DLAC),發(fā)展了一種新型的蛋白質(zhì)色譜復(fù)性新技術(shù)。采用具有表面大孔結(jié)構(gòu)的殼聚糖-硅基(CS-silica)材料作為DLAC的基質(zhì)材料。利用基質(zhì)中的氨基,共價偶聯(lián)親和染料配基活性藍(lán)(CBF)得到DLAC吸附劑。模型蛋白質(zhì)過氧化氫酶經(jīng)6 mol/L尿素變性后,DLAC可迅速去除變性劑,并通過CBF-CS-silica的吸附成功實現(xiàn)過氧化氫酶的復(fù)性。利用熒光光譜和過氧化氫酶活性研究考察了變性過程并對色譜洗脫步驟進(jìn)行優(yōu)化。與常規(guī)吸附復(fù)性法相比,可在蛋白質(zhì)濃度提高20倍的條件下進(jìn)行DLAC復(fù)性。 2、提出了制備新型有機-無機復(fù)合基質(zhì)的有效途徑。以球形硅膠作為支載核、以殼聚糖有機-無機雜化層作為殼,基于溶膠-凝膠反應(yīng),通過氨水簡單處理后制備了該吸附劑。在基質(zhì)上固定金屬離子后,即得到吸附蛋白質(zhì)的固定化金屬親和吸附劑。掃描電鏡表征表明,經(jīng)氨水溶液簡單處理后,基質(zhì)具有穩(wěn)定的多孔表面。X-衍射分析表明,有機-無機雜化和氨水處理過程可破壞殼聚糖的結(jié)晶區(qū),從而提高活性氨基的可利用度。采用同步熱重-微量熱分析進(jìn)一步對制備的基質(zhì)進(jìn)行了表征。在基質(zhì)表面固定作為特異性親和配體Cu2+后,制備了新型固定化金屬親和吸附劑。以BSA為模型蛋白,通過吸附實驗考察了該吸附劑的吸附性能。該吸附劑對BSA吸附速度快、吸附容量大。提出的方法及制備的基質(zhì)在生化分析中具有潛在的應(yīng)用前景。 3、提出了制備新型有機-無機生物吸附劑的有效途徑。以球形硅膠作為支載核、以殼聚糖有機-無機雜化層作為殼,基于通過溶膠-凝膠反應(yīng),經(jīng)氫氧化鈉溶液簡單處理后制備了該生物吸附劑。有機-無機雜化層通過殼聚糖與前驅(qū)體γ-環(huán)氧丙氧丙基三甲氧基硅烷(GPTMS)溶膠-凝膠化過程共價固定到硅膠表面。CS上的氨基與GPTMS上的環(huán)氧基團(tuán)交聯(lián)后,克服了其酸溶性缺點。通過氫氧化鈉溶液簡單處理后發(fā)生的濕法相轉(zhuǎn)移,使制備的復(fù)合生物吸附劑具有粗糙的表面。制備的生物吸附試劑可以用來處理電鍍廢水。
[Abstract]:This thesis is divided into three parts. In the first part, a new protein chromatography renaturation technique based on macroporous biomaterial dye affinity chromatography is developed, the second part is based on sol-gel method. The immobilized metal affinity adsorbent supported by spherical silica gel was prepared by the simple treatment of ammonia solution. The third part, based on the sol-gel method, the spherical silica supported biological adsorbent was prepared by the simple treatment of sodium hydroxide solution. 1. A novel protein chromatographic renaturation technique was developed based on dye affinity chromatography (DLA) of macroporous biomaterials. Chitosan / Si-based CS-silicon material with surface macroporous structure was used as the matrix material of DLAC. The model protein catalase was denatured by 6 mol/L urea, and the denaturant was removed rapidly. The renaturation of catalase was successfully achieved by the adsorption of CBF-CS-silica. The denaturation process was investigated by fluorescence spectrum and catalase activity. The chromatographic elution procedure was optimized. DLAC renaturation could be carried out under the condition of increasing protein concentration by 20 times. 2. An effective way to prepare a novel organic-inorganic composite matrix was proposed. The spherical silica gel was used as the supporting nucleus, the chitosan organic-inorganic hybrid layer was used as the shell, and the sol-gel reaction was carried out. The adsorbent was prepared by simple treatment of ammonia water. After immobilized metal ions were fixed on the substrate, the immobilized metal affinity adsorbent of adsorbed protein was obtained. The SEM characterization showed that the adsorbent was simply treated with ammonia solution. The matrix has a stable porous surface. X- diffraction analysis shows that organic-inorganic hybrid and ammonia treatment can destroy the crystalline region of chitosan. In order to improve the availability of active amino groups, the prepared matrix was further characterized by simultaneous thermogravimetry and microcalorimetry. After the substrate surface was fixed as a specific affinity ligand Cu2, A new immobilized metal affinity adsorbent was prepared. The adsorption performance of the immobilized metal affinity adsorbent was investigated by adsorption experiments using BSA as the model protein. The proposed method and the prepared matrix have potential application in biochemical analysis. 3. An effective way to prepare new organic-inorganic biological adsorbents was proposed. The spherical silica gel was used as the supporting nucleus, and the chitosan organic-inorganic hybrid layer was used as the shell, based on the sol-gel reaction. The biological adsorbent was prepared by simple treatment of sodium hydroxide solution. The organic-inorganic hybrid layer was covalently immobilized on the surface of silica gel through the sol-gelation process between chitosan and the precursor 緯 -epichloropropyltrimethoxysilane (GPTMS). When the amino group on CS is crosslinked with the epoxy group on GPTMS, Through the wet phase transfer after simple treatment of sodium hydroxide solution, the composite biological adsorbent has rough surface. The prepared biosorption reagent can be used to treat electroplating wastewater.
【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：Q51-3;R341

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