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复合纤维素膜固定化胰蛋白酶反应器及其应用于蛋白质酶解 被引量:17
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作者 姜泓海 邹汉法 +2 位作者 汪海林 倪坚毅 张强 《高等学校化学学报》 SCIE EI CAS CSCD 北大核心 2000年第5期702-706,共5页
合成了甲基丙烯酸缩水甘油酯-纤维素复合膜,并以此膜为基质共价键合固定化胰蛋白酶,以N-苯甲酰-L-精氨酰乙酯(BAEE)为底物,应用高效液相色谱系统测定了酶固定化膜柱的催化反应特性.研究结果表明:温度、pH值、离子强... 合成了甲基丙烯酸缩水甘油酯-纤维素复合膜,并以此膜为基质共价键合固定化胰蛋白酶,以N-苯甲酰-L-精氨酰乙酯(BAEE)为底物,应用高效液相色谱系统测定了酶固定化膜柱的催化反应特性.研究结果表明:温度、pH值、离子强度、有机溶剂及蛋白变性剂等都对固定化酶的活力有一定的影响.在最适条件下,固定化胰蛋白酶的活力为 17 800 U/g干膜,蛋白载量为 3. 6 mg/g(≈0. 15 μmol/g)干膜,活性回收率达到 52%.固定化酶表现出较高的使用和储藏稳定性,在40℃下,水解BAEE底物24 h活力无显著变化.固定化酶膜柱在4 ℃冷藏保存 100 d仍保存 90%以上的水解活力.固定化酶反应器被应用于蛋白质酶解的肽谱实验. 展开更多
关键词 复合纤维素膜 固定化胰蛋白酶反应器 蛋白质
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Combination of MALDI-TOF mass spectrometry with immobilized enzyme microreactor for peptide mapping 被引量:1
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作者 姜泓海 邹汉法 +5 位作者 汪海林 张强 倪坚毅 张清春 郭忠 陈小明 《Science China Chemistry》 SCIE EI CAS 2000年第6期625-633,共9页
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been combined with immobilized enzyme microreactor for the rapid, sensitive, and accurate tryptic mapping of protein and polypeptides. The t... Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been combined with immobilized enzyme microreactor for the rapid, sensitive, and accurate tryptic mapping of protein and polypeptides. The technique utilizes the trypsin microreactor by immobilized enzyme on the glycidyl methacrylate (GMA)-modified cellulose membrane. The membrane micro-reactor was used for the tryptic mapping of cytochrome C and the results were compared with those obtained by using free trypsin. A significant increase in the overall sensitivity of the process was observed using the membrane microreactor, as well as the elimination of background signals due to the autolysis of the trypsin. Further, membrane microreactor digestions were found to be rapid and convenient. 展开更多
关键词 IMMOBILIZED ENZYME MICROREACTOR glycidyl methacrylate (gma)-modified cellulose membrane MALDI-TOF MS peptide mapping.
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