| 壳聚糖层析凝胶洗脱条件优化 |
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| 引用本文： | 唐振兴,钱俊青,石陆娥.壳聚糖层析凝胶洗脱条件优化[J].精细化工,2004,21(10):726-730. |
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| 作者姓名： | 唐振兴 钱俊青 石陆娥 |
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| 作者单位： | 1. 浙江工业大学,化学工程与材料学院,浙江,杭州,310014
2. 浙江工业大学,药学院,浙江,杭州,310014 |
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| 摘 要： | 以壳聚糖为原料,经交联还原,制备了一种新型层析凝胶。为更好地应用该层析凝胶,对其洗脱条件进行了优化。确定了最佳洗脱体系是三羟甲基氨基甲烷(以下简称为tris HCl)溶液,体系的pH=9 05。考察了层析凝胶的颗粒度、洗脱液流速、离子强度、pH等条件对分离效果的影响,得到壳聚糖生物层析凝胶洗脱条件为:120～140μm壳聚糖层析凝胶装柱,用c(NaCl)=0 05mol/L的tris HCl(pH=9 05)洗脱,控制2 0～3 0mL/min流速。中性蛋白酶经壳聚糖层析凝胶一次层析,可分离得到4个组分,总酶活收率达90%以上,而用SepharoseCL-6B的对比实验只获得一个组分,酶活收率只有43 11%。
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| 关 键 词： | 壳聚糖 层析凝胶 蛋白质 酶 |
| 文章编号： | 1003-5214(2004)10-0726-05 |
Optimization of Elution Conditions of Chitosan Chromatographic Gel |
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| TANG Zhen-xing,QIAN Jun-qing,SHI Lu-e.Optimization of Elution Conditions of Chitosan Chromatographic Gel[J].Fine Chemicals,2004,21(10):726-730. |
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| Authors: | TANG Zhen-xing QIAN Jun-qing SHI Lu-e |
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| Affiliation: | TANG Zhen-xing~1,QIAN Jun-qing~2,SHI Lu-e~2 |
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| Abstract: | A novel chromatographic gel was prepared from chitosan through gelation,cross-linking and reduction.In order to make use of this gel,the elution conditions were optimized as follows.The best eluent system was tris-HCl(pH=9.05).Particle size of chitosan chromatographic gel was 120～140 μm.The washing fluid was c(NaCl)=0.05 mol/L tris-HCl(pH=9.05) at flowrate of 2.0～3.0 mL/min.Neutral protease could be separated well through this chromatographic gel column into four ingredients with total activity above 90%,by contrast,separation with Sepharose CL-6B got only one peak with 43.11% yield of activity. |
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| Keywords: | chitosan chromatography gel protein enzyme |
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