| 盐析和双水相萃取法纯化细菌木聚糖酶的比较 |
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| 引用本文： | 汤回花,滕超,刘丹蕾,鹿发展.盐析和双水相萃取法纯化细菌木聚糖酶的比较[J].食品工业科技,2018,39(12):92-97. |
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| 作者姓名： | 汤回花 滕超 刘丹蕾 鹿发展 |
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| 作者单位： | 1. 食品质量与安全北京实验室, 北京工商大学食品学院, 北京 100048;2. 北京市食品添加剂工程技术研究中心, 北京工商大学食品学院, 北京 100048 |
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| 基金项目： | 国家重点研发计划(2017YFD0400206)。国家自然科学基金项目(31671793,31201449) |
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| 摘 要： | 通过对比硫酸铵盐析和双水相萃取两种木聚糖酶初级纯化方法的纯化效率,确定更加适于Paenibacillus sp.B1709木聚糖酶初级纯化的方法。结果显示,双水相萃取较盐析具有更好的木聚糖酶纯化回收效果,并对其主要实验参数进行优化,得出双水相萃取最佳条件为:聚乙二醇(PEG)4000 Da,浓度22%,最佳成相盐(NH4)2SO4,最适浓度18%,最适pH6.0,NaCl 2%。在此条件下,可实现分配系数9.90,酶活力回收率为90.84%,纯化倍数6.13。实验结果显示双水相萃取相较于盐析在大规模纯化尤其针对复杂纯化目标时具备一定技术的优势。
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| 关 键 词： | 硫酸铵盐析 双水相法 木聚糖酶 纯化 |
| 收稿时间： | 2017-11-24 |
Compare the efficiency of salting-out and aqueous two-phase on purification of bacterial xylanase |
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| TANG Hui-hua,TENG Chao,LIU Dan-lei,LU Fa-zhang.Compare the efficiency of salting-out and aqueous two-phase on purification of bacterial xylanase[J].Science and Technology of Food Industry,2018,39(12):92-97. |
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| Authors: | TANG Hui-hua TENG Chao LIU Dan-lei LU Fa-zhang |
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| Affiliation: | 1. Beijing Laboratory of Food Quality and Safety, Beijing Technology & Business University(BTBU), Beijing 100048, China;2. Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology & Business University(BTBU), Beijing 100048, China |
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| Abstract: | This study aimed to compare the efficiency of salting-out method and aqueous two-phase method in the separation and purification of xylanase from Paenibacillus sp. B1709,and determined the optimal extration methods. A higher recovery of xylanase purification was obtained by the aqueous two-phase extraction and parameters were optimized,the optimum concentrations of PEG4000,(NH4)2SO4,NaCl and pH were 22%,18%,2% and 6,respectively. Under these conditions,the results were achieved as follows:partition coefficient 9.90,enzyme activity recovery 90.84% and the purification factor 6.13.Therefore,the aqueous two-phase has an advantage of a certain technology in the large-scale purification especially for complex purification. |
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