| 化学添加剂提高重组α-环糊精葡萄糖基转移酶酶制剂稳定性 |
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| 引用本文： | 郑贤良,吴丹,李兆丰,陈坚,吴敬.化学添加剂提高重组α-环糊精葡萄糖基转移酶酶制剂稳定性[J].生物工程学报,2011,27(2):185-195. |
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| 作者姓名： | 郑贤良 吴丹 李兆丰 陈坚 吴敬 |
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| 作者单位： | 江南大学,食品科学与技术国家重点实验室,无锡,214122;江南大学生物工程学院,江南大学工业生物技术教育部重点实验室,无锡,214122 |
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| 基金项目： | 国家杰出青年基金 (No. 20625619)，食品科学与技术国家重点实验室科研基金 (No. SKLF-MB-200802)，国家重点研究发展计划 (973计划) (No. 2007CB 714036) 资助。 |
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| 摘 要： | 通过向重组α-环糊精葡萄糖基转移酶 (α-CGT酶) 液中添加化学添加剂以提高其热稳定性及贮存稳定性。在不同温度下研究了添加剂对酶液的贮存稳定性影响，并用圆二色谱 (CD) 研究了CGT酶在近紫外区和远紫外区蛋白质结构与热稳定性的变化关系。当单独加入各种添加剂在50 ℃水浴1 h和室温放置108 d后，发现含有20％甘油的酶液稳定性最好，与未加任何添加剂的对照酶液相比仍有91％和50％的酶活，对照酶液在50 ℃水浴1 h后仅有小于10％的活性，室温放置108 d后已经没有酶活。明胶、CaCl2和PEG40
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| 关 键 词： | α-环糊精葡萄糖基转移酶，化学添加剂，热稳定性，圆二色光谱 |
| 收稿时间： | 6/3/2010 12:00:00 AM |
| 修稿时间： | 2010/9/25 0:00:00 |
Enhanced storage stability of recombinant enzyme preparation of alpha-CGTase from Paenibacillus macerans by chemical additives |
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| Xianliang Zheng,Dan Wu,Zhaofeng Li,Jian Chen and Jing Wu.Enhanced storage stability of recombinant enzyme preparation of alpha-CGTase from Paenibacillus macerans by chemical additives[J].Chinese Journal of Biotechnology,2011,27(2):185-195. |
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| Authors: | Xianliang Zheng Dan Wu Zhaofeng Li Jian Chen and Jing Wu |
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| Institution: | State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China |
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| Abstract: | To enhance the thermostability and storage stability of alpha-cyclodextrin glycosyltransferase (a-CGTase), we added specific chemical additives into the preparation of alpha-CGTase, and studied the effect of additives on the storage stability of alpha-CGTase at different temperatures. Then we measured the protein structure of CGTase in the far UV (200-250 nm) and near UV (250-320 nm) ranges respectively by Circular dichroism (CD) spectra under high temperature and analyzed the relationship between thermostability and protein structure. The results indicated that the addition of selected additives (gelatin, glycerin, CaCl2 and PEG400) enhanced the thermostability of alpha-CGTase dramatically. After 45 days, the preparation of alpha-CGTase still had 100% of the enzyme activity with different additives superimposed at the optimum concentration at 40 degrees C. The CD spectra of alpha-CGTase showed that glycerin could protect the secondary and the tertiary structure of the CGTase under high temperature and therefore the enzyme maintained its high activity. Chemical additives can improve the stability of alpha-CGTase significantly and they preserve the enzyme activity by protecting its secondary structure. |
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| Keywords: | a-cyclodextrin glycosyltransferase chemical additives thermostability circular dichroism |
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