介孔分子筛SBA-15的脂肪酶固定量分析测定  

Detection of Amount of Lipase Immobilized on Molecular Sieve Mesoporous Silica SBA-15

作　　者：尚雁[1] 李娜[1] 覃小焕[1] 鲁保旺[2] 傅强[3] 

机构地区：[1]西安交通大学能动学院化工系,西安710049 [2]日本产业技术综合研究所(AIST) [3]西安交通大学医学院药学系,西安710061

出　　处：《分析化学》2009年第8期1173-1177,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.20406014);国家重点基础研究计划(973项目)(No.2009CB2199006)资助项目

摘　　要：利用吸附法将假丝酵母脂肪酶(candida rugosa lipase,CRL)固定在介孔分子筛SBA-15上,对比了由单波长紫外分光光度法、双波长紫外分光光度法和二辛可宁酸法(bicinchoninic acid method,BCA)法测定的酶蛋白浓度及酶蛋白固定量。结果表明:SBA-15对紫外吸收有明显干扰,单波长紫外法测定结果远大于双波长紫外法和BCA法,双波长紫外法和BCA法测定结果较接近。利用BCA法测定了不同浓度CRL在介孔分子筛上的固定量,考察了固定化酶的泄漏量。在编号分别为Lu001和LLSD1的介孔分子筛SBA-15上的载酶量分别为16.6和114.12mg/g。在缓冲溶液中SBA-15固定化酶的泄漏率只约为0.5%,可作为良好的酶固定化载体。Candida rugosa lipase（CRL） was immobilized on molecular sieve mesoporous silica SBA-15 by adsorption method. The concentration and the amount of immobilized enzyme protein were determined by three detection methods for comparison, including single wavelength UV speetrophotometrie method, double-wavelength UV speetrophotometrie method and bicinehoninie acid method（BCA）. It was found that the UV speetrophotometric method was interfered easily by SBA-15, and the single wavelength UV method gave much higher results than those by double-wavelength UV and BCA methods, which were relatively near with each other. In addition, the amounts of immobilized enzyme protein on SBA-15 were also measured by BCA method in the presence of various initial enzyme concentrations. The leakage of immobilized enzyme from SBA-15 into buffer solution was investigated through tracking measurement. The amounts of immobilized enzyme protein on two different molecular sieves mesoporous silica SBA-15, named as LLSD1 and Lu001, were 16.6 mg/g support and 114.2 mg/g support, respectively. The leakage amount of immobilized enzyme protein in buffer solution is only 0. 5% of total loading. The results prove that SBA-15 can be a feasible immobilization support for lipase.

关 键 词：介孔分子筛SBA-15 脂肪酶 酶蛋白固定量 紫外分光光度法 二辛可宁酸法 

分 类 号：O629.8[理学—有机化学]