催化学报 ›› 2014, Vol. 35 ›› Issue (4): 553-559.DOI: 10.1016/S1872-2067(14)60053-X

• 研究论文 • 上一篇    下一篇

紫外分光光度法表征Lipozyme TL IM脂肪酶转酯化活性

付显锋, 郑建永, 应向贤, 鄢洪德, 汪钊   

  1. 浙江工业大学生物与环境工程学院, 浙江杭州310032
  • 收稿日期:2013-11-15 修回日期:2014-01-07 出版日期:2014-03-20 发布日期:2014-03-21
  • 通讯作者: 汪钊
  • 基金资助:

    国家高技术研究发展计划(863计划,2011AA02A210).

Investigation of Lipozyme TL IM-catalyzed transesterification using ultraviolet spectrophotometric assay

Xianfeng Fu, Jianyong Zheng, Xiangxian Ying, Hongde Yan, Zhao Wang   

  1. College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, Zhejiang, China
  • Received:2013-11-15 Revised:2014-01-07 Online:2014-03-20 Published:2014-03-21
  • Supported by:

    This work was financially supported by the National High Technology Research and Development Program of China (863 Program, 2011AA02A210).

摘要:

建立了一种新的有机相脂肪酶转酯化活性测定方法. 以正己烷为溶剂,脂肪酶催化棕榈酸对硝基苯酯和正丁醇的转酯化反应为模型反应,通过测定反应液中310 nm下吸光值的变化计算反应转化率. 以气相色谱法对新建的紫外分光光度法进行验证,分别采用这两种方法测定了七种商品化脂肪酶的转酯化活性,两种方法所得实验结果基本一致. 利用紫外分光光度检测法考察了Lipozyme TL IM脂肪酶催化转酯化的时间进程及合成活性与酶量的关系,并对Lipozyme TL IM催化转酯化的性质(最适溶剂、酰基受体特异性、醇耐受性、最优反应温度和热力学稳定性)进行了表征.

关键词: 脂肪酶, Lipozyme TL IM, 转酯化活性, 紫外分光光度法, 棕榈酸对硝基苯酯, 有机溶剂

Abstract:

A novel ultraviolet (UV) spectrophotometric method for evaluating lipase transesterification activity in organic solvents is described. The model reaction was lipase-catalyzed transesterification between p-nitrophenyl palmitate (pNPP) and n-butanol in n-hexane at 30 ℃. The conversion of pNPP to p-nitrophenol was monitored by measuring UV absorbance changes at 310 nm. Seven commercial lipases were tested using this method, and the results were similar to those obtained using gas chromatography. The UV spectrophotometric method was used to investigate the kinetics of the Lipozyme TL IM-catalyzed reaction of pNPP, and the correlation between synthetic activity and biocatalyst concentration was determined. The method was also used to investigate the Lipozyme TL IM-catalyzed transesterification properties, namely solvent effect, acyl acceptor specificity, alcohol tolerance, optimum reaction temperature, and thermostability.

Key words: Lipase, Lipozyme TL IM, Transesterification activity, Ultraviolet spectrophotometric assay, p-Nitrophenyl palmitate, Organic solvent