催化学报

催化学报 ›› 2007, Vol. 28 ›› Issue (2): 175-179.

• 研究论文 • 上一篇    下一篇

粘质沙雷氏菌脂肪酶的固定化及催化拆分反式3-(4’-甲氧苯基)缩水甘油酸甲酯

龙章德1,2,许建和1,潘江1   

  1. 1 华东理工大学生物反应器工程国家重点实验室, 上海 200237; 2 广西大学水稻研究所, 广西南宁 530004
  • 收稿日期:2007-02-25 出版日期:2007-02-25 发布日期:2011-01-28

Immobilization of Serratia marcescens Lipase and Catalytic Resolution of Trans-3-(4’-methoxyphenyl)glycidic Acid Methyl Ester

LONG Zhangde1,2, XU Jianhe1*, PAN Jiang1   

  1. 1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China; 2 Rice Research Institute, Guangxi University, Nanning 530004, Guangxi, China
  • Received:2007-02-25 Online:2007-02-25 Published:2011-01-28

PDF

900

摘要： 对粘质沙雷氏菌脂肪酶进行了固定化研究,确定硅藻土和环氧树脂Eupergit C是较好的固定化载体. 固定化后酶的热稳定性、 pH稳定性及储存稳定性均明显提高. 以Eupergit C共价固定的脂肪酶,其操作稳定性比硅藻土吸附酶好,重复使用10批次后,剩余酶活力还有50%左右,戊二醛交联对Eupergit C固定化酶稳定性的提高没有明显效果,而硅藻土吸附酶经戊二醛交联后稳定性有所提高,经5批次反应后剩余酶活力还有50%. 使用交联后的硅藻土固定化酶(1 g, 200 U), 在两相搅拌反应器(工作体积200 ml, 甲苯∶水体积比=1)中对地尔硫卓手性前体(±)-反式3-(4’-甲氧苯基)缩水甘油酸甲酯((±)-MPGM)(有机相浓度为0.5 mol/L)进行了催化拆分,经5批次反应后,共得纯(2R,3S)-(-)-MPGM 18.6 g, 产品光学纯度(对映体过量)>99%, 总收率为37.2%.

关键词: 粘质沙雷氏菌, 脂肪酶, 固定化, 对映选择性水解, 反式3-(4&rsquo, -甲氧苯基)缩水甘油酸甲酯, 硅藻土, 环氧树脂, Eupergit C

Abstract: Seven carriers were investigated for immobilization of the lipase from Serratia marcescens ECU1010, and diatomite and Eupergit C were found to be ideal supports. The thermal and storage stability of the lipase was significantly improved after being immobilized onto these two supports. After heating at 50 ℃ for 6 h, only a small decrease in the activity was observed for the diatomite-adsorbed lipase. About 50% of the initial activity was retained when the two immobilized enzyme samples were storaged at 4 ℃ for 180 days. Operational stability tests indicated that the half-life of the lipase immobilized on Eupergit C was about 2-fold longer than that of the diatomite-immobilized lipase. The crosslinking with glutaraldehyde had no significant effect on the activity of the Eupergit C-immobilized lipase, whereas the reusability of the crosslinked diatomite-immobilized enzyme was slightly improved. Using the crosslinked diatomite-immobilized lipase, an up-scaled bioresolution of (±)-3-(4’-methoxyphenyl)glycidic acid methyl ester ((±)-MPGM) was performed in a 500-ml emulsion reactor with 200 ml biphasic solution (toluene∶water=1∶1) at 30 ℃ and 0.5 mol/L of (±)-MPGM in toluene. After 5 batches of reaction, crystalline (2R,3S)-(-)-MPGM with a chemical purity of 100% and optical purity of>99% enantiomeric excess was obtained in a yield of 37.2%.

Key words: Serratia marcescens, lipase, immobilization, enantioselective hydrolysis, 3-(4’-methoxyphenyl)glycidic acid ester, diatomite, epoxy resin, Eupergit C