催化学报

催化学报 ›› 2011, Vol. 32 ›› Issue (4): 630-636.DOI: 10.3724/SP.J.1088.2011.01129

• 研究论文 • 上一篇    下一篇

核苷酸锆固定化脂肪酶的制备及其催化性能

陶维红, 杨立荣, 徐刚, 乔元彪, 吴坚平   

  1. 浙江大学化学工程与生物工程学系, 浙江杭州 310027
  • 收稿日期:2010-11-29 修回日期:2011-01-17 出版日期:2011-04-18 发布日期:2014-08-30

The Immobilization of Lipase on Zirconium Nucleotide and Its Catalytic Performance

TAO Weihong, YANG Lirong, XU Gang, QIAO Yuanbiao, WU Jianping*   

  1. Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, Zhejiang, China
  • Received:2010-11-29 Revised:2011-01-17 Online:2011-04-18 Published:2014-08-30

PDF

918

被引次数

14

摘要： 开发了以核苷酸锆纳米颗粒为载体固定化脂肪酶的方法, 考察了固定化时间、缓冲液 pH 值及浓度和酶载量等因素对固定化酶在有机相中催化烯丙醇酮酯交换反应性能的影响. 结果表明, 一元尿苷酸锆 (粒径 30~50 nm) 是较佳的载体, 在磷酸缓冲液 pH = 8.0 和浓度为 0.03 mol/L 时, 酶与载体质量比为 4, 固定化 6 h 制得的固定化酶效果最佳. 固定化酶的活力回收率是游离酶的 6.8 倍; 对映体选择率 (E = 200) 明显高于游离酶 (E = 86). 以一元尿苷酸锆作为载体制得的固定化酶的操作稳定性显著提高, 重复利用 20 批次后残余活力为 58.7%, 而游离酶仅为 4%.

关键词: 核苷酸锆, 纳米颗粒, 节杆菌脂肪酶, 固定化, 酯交换反应

Abstract: The immobilization of Arthrobacter sp. lipase on zirconium nucleotide nanoparticles was investigated. The effect of immobilization time, pH value, buffer concentration, and lipase amount on the catalytic performance of the immobilized lipase in an organic system was studied. The results showed that the enzyme immobilized on zirconium-uridine monophosphate (30–50 nm) had the best catalytic performance for the resolution of 4-hydroxy-3-methyl-2-(2-propenyl)-2-cyclopenten-1-one. The optimal immobilization conditions were determined to be pH = 8.0, phosphate buffer concentration of 0.03 mol/L with the lipase/support mass ratio of 4 , and immobilization for 6 h. The activity recovery was as high as 6.8 fold of the free lipase and the enantioselectivity (E = 200) was improved (E = 85 for free lipase). Furthermore, the operational stability for the immobilized enzyme was greatly enhanced compared with the free enzyme. The residual activity was kept at 58.7% after 20 batch reactions, while that of the free enzyme was only 4%.

Key words: zirconium nucleotide, nanoparticle, Arthrobacter sp. lipase, immobilization, transesterification