以邻苯二甲酸二丁酯为致孔剂,制备多孔壳聚糖膜,用作固定化脲酶的载体.利用X射线微区分析方法,对其活性进行了定位分析:以BaCl2为捕捉剂,在Tris-HCl缓冲溶液中,底物尿素经固定化脲酶催化水解产生NH3和CO2,后者和捕捉剂反应生成BaCO3,沉积在固定化脲酶的催化活性部位.结果表明:BaCl2可用作固定化脲酶活性定位的捕捉剂;多孔壳聚糖膜通透性好、比表面积大、载酶量高,脲酶均匀分布在膜的外表面和膜内的孔隙中.
[1] 李沁华, 孙娜. 多孔壳聚糖膜材料的制备及性能研究 [J]. 生物医学工程学杂志 1999.doi:10.3321/j.issn:1001-5515.1999.Z1.047
[2] 姚子华, 黄永章, 仇满德, 王书香, 张贺迎, 武金霞, 潘延云, 周艳芬, YAO Zi-hua, HUANG Yong-zhang, QIU Man-de, WANG Shu-xiang, ZHANG He-ying, WU Jin-xia, PAN Yan-yun, ZHOU Yan-fen. 固定化青霉素G酰化酶活性的X射线微分析(Ⅱ)-活性定位 [J]. 河北大学学报(自然科学版) 2000.doi:10.3969/j.issn.1000-1565.2000.04.020
[3] 黄永章, 姚子华, 王桂华, 仇满德. 固定化葡萄糖氧化酶活性的X射线微区分析 [J]. 分析化学 2001.doi:10.3321/j.issn:0253-3820.2001.12.018
[4] 马晓莉, 姚子华, MA Xiao-li, YAO Zi-hua. 壳聚糖固定化脲酶活性的X射线微区分析 [J]. 光谱学与光谱分析 2005.doi:10.3321/j.issn:1000-0593.2005.03.036
[5] Ruckenstein E., Zeng XF.. Albumin separation with Cibacron Blue carrying macroporous chitosan and chitin affinity membranes [J]. Journal of Membrane Science 1998, 1(1).
[6] Eli Ruckenstein, XIANFANG ZENG. Macroporous chitin affinity membranes for Lysozyme separation [J]. Biotechnology and Bioengineering 1997, 56.
[7] Ma J, Wang H, He B, Chen J. A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetal dermal fibroblasts. [J]. Biomaterials 2001, 4(4).
[8] Mi FL, Shyu SS, Wu YB, Lee ST, Shyong JY, Huang RN. Fabrication and characterization of a sponge-like asymmetric chitosan membrane as a wound dressing. [J]. Biomaterials 2001, 2(2).
[9] Krajewska B, ZABORSKA W. Dialysate regeneration system of artificial kidney [J]. Biocybernetics and Biomedical Engineering 1989, 9.
[10] George S, CHELLAPANDIAN M, SIVASANKAR B. A new process for the treatment of fertilizer effluent using immobilized urease [J]. Biomaterials 1995, 16.
[11] Li O, DUNN E T, GRANDMAISON E W. Applications and properties of chitosan [J]. Journal of Bioactive and Compatible Polymers 1992, 7.
[12] Julia M C S. Urea potentiometric biosensor based on urease imnobilized on chitosan membranes [J]. Talanta 1998, 47.
[13] ROOSEVEAR A, KENNEDY J F, CABRAL J M S. Immobilized enzymes and cells [M]. Bristol:Adam Hilger 1987.
[14] WEATHERBURN M W. Phenol-hypochlorite reaction for determination of ammonia [J]. Analytical Chemistry 1967, 39.
[15] 蒋挺大. 甲壳素 [M]. 北京:化学工业出版社 2003.