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聚甲基戊烯中空纤维膜表面碳酸酐酶的固定化及表征

[Immobilization and characterization of carbonic anhydrase on the surface of hollow fiber membrane of polymethyl pentene].

作者信息

Wang Qinmei, Zhang Dihua, Zhang Jingxia

机构信息

First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2009 Jul;25(7):1055-61.

Abstract

We immobilized carbonic anhydrase (CA) onto the surface of membrane oxygenator of polymethyl pentene (PMP) to enhance the removal of carbon dioxide in blood by two steps. We first introduced hydroxyl groups onto PMP surface by water plasma treatment, and then coupled CA onto PMP surface by using cyanate bromide (CNBr) as a crosslinker. After plasma treatment, the contact angle with water and chemical composition of PMP surface were characterized by analysis system of surface contact angle and XPS. Using p-nitrophenyl acetate (p-NPA) as a substrate, the activity, concentration, storage stability and re-usability of immobilized CA on PMP hollow fibers were studied by ultraviolet spectrophotometer. The preliminary data showed that hydroxyl groups could be introduced on the surface of PMP by water plasma treatment, and CA with catalysis activity could be successfully introduced onto PMP surface in high immobilization efficiency. The activity of covalently immobilized CA increased with the increase of concentration of CNBr, and the maximum was 73% of the theoretical activity of CA spread on PMP surface in monolayer in studied range. Covalently immobilized CA showed higher reusability compared to physically adsorbed CA, and higher storage stability compared to free CA in solution at 37 degrees C. The method would be used potentially in the membrane oxygenator to improve the capacity of removal of carbon dioxide in blood in the future.

摘要

我们通过两步法将碳酸酐酶(CA)固定在聚甲基戊烯(PMP)膜式氧合器表面,以增强血液中二氧化碳的清除。我们首先通过水等离子体处理在PMP表面引入羟基,然后使用溴化氰(CNBr)作为交联剂将CA偶联到PMP表面。等离子体处理后,通过表面接触角分析系统和XPS对PMP表面与水的接触角和化学成分进行了表征。以对硝基苯乙酸(p-NPA)为底物,用紫外分光光度计研究了固定在PMP中空纤维上的CA的活性、浓度、储存稳定性和可重复使用性。初步数据表明,通过水等离子体处理可在PMP表面引入羟基,并且具有催化活性的CA能够以高固定效率成功引入到PMP表面。共价固定的CA的活性随CNBr浓度的增加而增加,在所研究的范围内,其最大值为单层铺展在PMP表面的CA理论活性的73%。与物理吸附的CA相比,共价固定的CA表现出更高的可重复使用性,与37℃溶液中的游离CA相比,具有更高的储存稳定性。该方法未来可能会用于膜式氧合器中,以提高血液中二氧化碳的清除能力。

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