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| 凝胶层析和离子交换层析结合法纯化高盐发酵液中谷氨酰胺转胺酶 | |
| 引用本文： | 张莉丽,张兰威,杜明,韩雪,易华西,张英春,张艳禾,马微.凝胶层析和离子交换层析结合法纯化高盐发酵液中谷氨酰胺转胺酶[J].分析化学,2012,40(8):1225-1230. |
| 作者姓名： | 张莉丽 张兰威 杜明 韩雪 易华西 张英春 张艳禾 马微 |
| 作者单位： | 1. 哈尔滨工业大学食品科学与工程学院,哈尔滨150090;东北农业大学食品学院,哈尔滨150030 2. 哈尔滨工业大学食品科学与工程学院,哈尔滨,150090 3. 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物细菌病研究室,哈尔滨,150001 |
| 基金项目： | 国家自然科学基金，黑龙江省自然科学基金，黑龙江省科技厅项目，哈尔滨市基金 |
| 摘 要： | 采用凝胶层析和离子交换层析相结合的方法分离纯化高盐培养基中的谷氨酰胺转胺酶,优化的凝胶层析的条件,上样量6mL,流速为0.25 mL/min；离子交换层析的上样量50 mL,流速为3 mL/min.酶被纯化了4.22倍,比活力达17.33 U/mg蛋白,回收率为77.5％.液相色谱-串联质谱鉴定、蛋白质数据库比对结果表明,纯酶与AAN01353是同种蛋白质. |
| 关 键 词： | 谷氨酰胺转胺酶 分离纯化 液相色谱-串联质谱 凝胶层析 离子交换层析 |
Purification of Transglutaminase from High Salt Culture by Gel and Cation Exchange Chromatography |
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| ZHANG Li-Li , ZHANG Lan-Wei , DU Ming , HAN Xue , YI Hua-Xi , ZHANG Ying-Chun , ZHANG Yan-He , MA Wei.Purification of Transglutaminase from High Salt Culture by Gel and Cation Exchange Chromatography[J].Chinese Journal of Analytical Chemistry,2012,40(8):1225-1230. | |
| Authors: | ZHANG Li-Li ZHANG Lan-Wei DU Ming HAN Xue YI Hua-Xi ZHANG Ying-Chun ZHANG Yan-He MA Wei |
| Affiliation: | 1(School of Food Science and Engineering,Harbin Institute of Technology,Harbin 150090,China) 2(Food College,Northeast Agricultural University,Harbin 150030,China) 3(National Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
| Abstract: | To purify the transglutaminase from high-salt medium,gel filtration combined with ion exchange chromatography method was employed.Two-step chromatographic conditions(flow rate and the sample volume) were optimized.The final sample volume and a flow rate for Gel chromatography were 6 mL and 0.25 mL/min,respectively.Conditions for ion-exchange chromatography were a sample volume of 50 mL and flow rate of 3 mL/min.Enzyme was purified 4.22-fold,the specific activity and the recovery rate were 17.33 U/mg protein and 77.5%,respectrively.A comparison between liquid chromatography/tandem mass spectrometric identification and the protein database was carried out and the results showed that the purified enzyme had high homology with AAN01353. |
| Keywords: | Transglutaminase Purification Liquid chromatography/tandem mass spectrometry Gel chromatography Cation exchange chromatography |
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