高等学校化学学报

高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (5): 1195.doi: 10.7503/cjcu20120951

• 物理化学 • 上一篇    下一篇

Burkholderia Cepacia脂肪酶在氨基功能化离子液体修饰的SBA-15上的共价固定

胡燚1,2, 杨姣1, 唐苏苏1, 初旭明1, 邹彬1, 黄和1,2   

  1. 1. 南京工业大学生物与制药工程学院, 南京 210009;
    2. 南京工业大学材料化学工程国家重点实验室, 南京 210009
  • 收稿日期:2012-10-22 出版日期:2013-05-10 发布日期:2013-05-10
  • 通讯作者: 黄 和, 男, 博士, 教授, 主要从事工业微生物的代谢工程及代谢组学方面的研究. E-mail: biotech@njut.edu.cn E-mail:biotech@njut.edu.cn
  • 基金资助:

    国家杰出青年科学基金(批准号: 21225626)、 国家自然科学基金(20906049, 20936002)和国家"八六三"计划项目(批准号: 2011AA02A209)资助.

Covalent Immobilization of Burkholderia Cepacia Lipase on Amine Functionalized Ionic Liquid Modified SBA-15

HU Yi1,2, YANG Jiao1, TANG Su-Su1, CHU Xu-Ming1, ZOU Bin1, HUANG He1,2   

  1. 1. College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China;
    2. State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing University of Technology, Nanjing 210009, China
  • Received:2012-10-22 Online:2013-05-10 Published:2013-05-10

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摘要：

合成了氨基以及氨基功能化离子液体修饰的介孔材料SBA-15(NH2-SBA和NH2-IL-SBA), 并以戊二醛为活化剂对NH2-IL-SBA进行活化处理(CA-NH2-IL-SBA), 通过元素分析、 N2吸附-脱附、 X射线衍射、 红外光谱等方法研究了修饰及活化对SBA-15结构的影响. 将所得新型固定化载体用于Burkholderia cepacia脂肪酶(BCL)的吸附固定、 共价交联固定及聚集包被固定. 以三乙酸甘油酯的水解为模型反应, 考察了固定化BCL的酶活、 最适反应条件、 稳定性等酶学性质. 结果表明, 离子液体修饰后的载体保持了原有的孔道结构, 与氨基修饰以及原粉SBA-15吸附固定的BCL(BCL-NH2-SBA和BCL-SBA-15)相比, 其固定化酶的比活力和稳定性都得到了明显提高, 对温度及低pH的敏感性降低. 其中聚集包被固定的BCL在获得了相对较高酶负载量的同时显示了最好的稳定性, 其热稳定性和重复使用性分别为BCL-SBA-15的4倍和2倍.

关键词: Burkholderia cepacia脂肪酶, 离子液体, 介孔材料, 修饰, 固定化

Abstract:

The (3-aminopropyl)trimethoxysilane and amine functionalized ionic liquid modified mesoporous silica SBA-15(NH2-SBA, NH2-IL-SBA) were synthesized, and NH2-IL-SBA was activated by glutaraldehyde(CA-NH2-IL-SBA). Physical and chemical properties of the materials were characterized by elemental analysis, nitrogen adsorption-desorption, small-angle X-ray diffraction and Fourier transform infrared spectroscopy. The prepared materials were used as novel carrier systems to immobilize Burkholderia cepacia lipase(BCL) by physical adsorption, conventional covalent attachment and enzyme-aggregate coating methods. Enzymatic properties, including activity, optimum reaction condition and stability were investigated in the triacetin hydrolysis reaction. The results showed that the modification and activation did not destroy the structure of SBA-15. Compared with the immobilized BCL on NH2-SBA and parent SBA-15(BCL-NH2-SBA, BCL-SBA-15), the immobilized BCL on novel carriers were more resistant to temperature and low pH changes and possessed both higher activity and stability. Especially, the BCL prepared by enzyme-aggregate coating method exhibited highest immobilization efficiency and stability, which improved 4.0 and 2.0 folds of thermal stability and reusability than BCL-SBA-15, respectively.

Key words: Burkholderia cepacia lipase, Ionic liquid, Mesoporous material, Modification, Immobilization

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