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Citation: WANG Qin-Mei, ZHANG Di-Hua, ZHANG Jing-Xia. Immobilization of Carbonic Anhydrase onto Polymer Surface without Active Functional Groups with Crosslinker[J]. Chinese Journal of Applied Chemistry, ;2009, 26(6): 637-641. shu

Immobilization of Carbonic Anhydrase onto Polymer Surface without Active Functional Groups with Crosslinker

1.
a Laboratory of Bioactive Materials, First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080;
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b Institute of Cardiology and Angiology, Sun Yat-sen University, Guangzhou 510080;
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c Center of Blood Dialysis, First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080;
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d School of Pharmaceutical Science, Sun Yat-sen University, Guangzhou 510080

Corresponding author: WANG Qin-Mei,
Received Date: 20 June 2008
Available Online: 18 September 2008
Fund Project: 教育部留学回国人员科研启动基金(20071108)资助项目 (20071108)

Carbonic anhydrase(CA) was immobilized onto the surface of hollow fiber membrane of polymethyl pentene(PMP) without any active functional groups via sulfosuccinimidyl [perfluoroazidobenzamido] ethyl-1,3-dithiopropionate(SFAD),a photoactivatable heterobifunctional crosslinker.Using p-nitrophenyl acetate(p-NPA) as a substrate,the activity,concentration,storage stability and re-usability of immobilized CA were studied by ultraviolet spectrophotometry.The preliminary data showed that CA could be successfully immobilized onto PMP surface;covalently immobilized CA(CA_CI) had catalysis activity.Its concentration on PMP surface increased with the increase of the SFAD concentration,and reached a maximum of 0.414 U/m²,which was 87.3% of the theoretical concentration of CA on PMP surface in monolayer,and the reusability was much better than physically adsorbed CA(CA_PA): the activity of CA_PA on PMP surface was 0 after washing once,but CA_CI retained around 59% of its activity after washing 9 times;CA_CI showed higher storage stability compared to free CA in solution at 37 ℃: the soluble enzyme lost about 90% activity,but CA_CI lost only 60% activity in 16 days.
- polymethyl pentene,
- photoreactive crosslinker,
- carbonic anhydrase,
- p-nitrophenyl acetate
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