催化学报

催化学报 ›› 2011, Vol. 32 ›› Issue (12): 1869-1874.DOI: 10.3724/SP.J.1088.2011.10536

• 研究论文 • 上一篇    下一篇

亲水梳状环氧聚合物载体柔性固定化脂肪酶

曲伟光, 魏荣卿, 何冰芳, 刘晓宁*, 仲玉, 陈新营   

  1. 南京工业大学生物与制药工程学院, 江苏南京 210009
  • 收稿日期:2011-07-17 修回日期:2011-09-07 出版日期:2011-12-21 发布日期:2015-04-23

Flexible Immobilization of Lipase on Hydrophilic and Comblike Polymer Support Containing Epoxy Group

QU Weiguang, WEI Rongqing, HE Bingfang, LIU Xiaoning*, ZHONG Yu, CHEN Xinying   

  1. College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, Jiangsu, China
  • Received:2011-07-17 Revised:2011-09-07 Online:2011-12-21 Published:2015-04-23

PDF

879

被引次数

48

摘要： 以氯乙酰化聚苯乙烯微球载体为大分子引发剂, 以甲基丙烯酸缩水甘油酯和亲水性丙烯酰胺为共聚接枝单体, 以氯化亚铜及 2,2′-联吡啶为催化体系, 采用原子转移自由基聚合法接枝合成了具有柔性链的亲水梳状环氧聚合物载体 PS-acyl-P(AM-co-GMA), 并将其用于耐有机溶剂 YCJ01 脂肪酶的共价柔性固定化. 结果表明, 固定化酶催化对硝基苯酚棕榈酸酯水解的最适温度为 45~55 oC, pH = 8.5, 此时酶活为 16.6 U/g; 在 80 oC 以下, pH = 5~10 时, 酶的活性稳定; 连续进行 7 次间歇操作, 固定化酶保留初始活性的 73.5%; 于 4 oC 保存 60 d, 酶活仍剩余 86.8%. 经亲水梳状环氧载体柔性固定化后, 该脂肪酶对有机溶剂的耐受性有所提高.

关键词: 氯乙酰化聚苯乙烯, 原子转移自由基聚合, 梳状环氧聚合物, 耐有机溶剂脂肪酶, 柔性固定化

Abstract: A hydrophilic and comblike polymer support PS-acyl-P(acrylamide-co-glycidyl methacrylate) containing epoxy groups for flexible and covalent immobilization of lipase has been prepared by an atom transfer radical polymerization method. The grafting reaction of glycidyl methacylate and hydrophilic acrylamid was initiated from the chloroacetyl groups on the styrene-divinylbenzene copolymer microspheres with CuCl and 2,2′-bipyridine complex as a catalyst. The apparent activity of lipase immobilized on PS-acyl-P(acrylamide-co-glycidyl methacrylate) was 16.6 U/g, and the optimal temperature and pH were 45–55 oC and 8.5 for hydrolysis of p-nitrophenyl palmitate, respectively. The immobilized lipase was stable in the range of pH = 5–10 and at temperature below 80 oC. The operational stability for the immobilized lipase was tested for consecutive hydrolysis of p-nitrophenyl palmitate. The result shows that the immobilized lipase has excellent operational stability. The residual activity of immobilized lipase was 73.5% after used 7 cycles. The activity of immobilized lipase remained 86.8% of the origin after storage at 4 oC for 60 d. Lipase got improved to varying degrees for organic solvent tolerance through immobilization on PS-acyl-P(acrylamide-co-glycidyl methacrylate).

Key words: chloroacetylated polystyrene, atom transfer radical polymerization, comblike polymer containing epoxy group, organic solvent-tolerant lipase, flexible immobilization