预混合自组装辣根过氧化物酶生物活性膜及其催化活性研究  

STUDY ON THE FABRICATION AND THE ACTIVITY OF HORSERADISH PEROXIDASE BIOFILMS SELF-ASSEMBLED BY PREMIXING METHOD

作　　者：江秀明[1] 陈志春[1] 杨绍明[1] 林贤福[1] 

机构地区：[1]浙江大学化学系

出　　处：《高分子学报》2005年第2期181-185,共5页Acta Polymerica Sinica

基　　金：浙江省科技厅国际重点合作项目 (项目号 0 2 110 613 2 )资助

摘　　要：以阳离子化的辣根过氧化物酶 (HRP)和阴离子聚苯乙烯磺酸钠 (PSS)的预混合溶液 ,与阳离子聚电解质聚二甲基二烯丙基氯化铵 (PDDA)通过逐层组装 ,在阴离子化聚对苯二酸乙二酯 (PET)表面构建了多层生物活性膜 .用紫外 可见光谱仪 (UV Vis)和原子力显微镜 (AFM)研究了交替自组装膜的结构和表面形膜 ,并测定了自组装膜的生物催化活性 .结果表明 ,预混合溶液中的PSS与HRP一起沉积在PDDA膜层上组装成 (PSS+HRP)膜层 ,且每层中PSS和HRP的比例一致 ;(PSS +HRP)膜层呈条状分布 ,膜表面较为平整 ;多层膜中的HRP催化H2 O2 与 4 氨基安替比林的显色反应的表观米氏常数为 9 7× 10 - 5mol·L- 1 (相对于H2 O2 底物 ) ,较溶液中 (1 5 2× 10 - 4mol·L- 1 )的小 .The premixing self-assembly solution of horseradish peroxide with poly(sodium 4-styrenesulfonate) (HRP + PSS) is made by blending HRP and polyelectrolyte PSS, and the active [(HRP + PSS)/PDDA](n) enzyme multilayer biofilm is fabricated by alternate deposition of ( HRP + PSS) and PDDA on the surface of negative ionized PET. The topography of films, the stepwise growth of layers and the activity of HRP in the film are measured with UV-Vis and AFM. HRP and PSS are assembled on the PDDA layer together from the premixing solution of ( HRP + PSS), and the ratio of HRP to PSS in every layer of (HRP + PSS) is the same. The banded structure of HRP under the PSS is observed on the PDDA layer. The Michaelis Constant of the reaction of H2O2, and 4-amino-antipyrine (4-AAP) catalyzed by HRP in biofilm is 9.7 x 10 mol(.)L and is lower than that of the reaction catalyzed by HRP in solution (1.52 x 10(-4) mol-L The affinity between H2O2, and HRP in biofilms is rather strong.

关 键 词：辣根过氧化物酶 生物活性膜 活性研究 聚二甲基二烯丙基氯化铵 原子力显微镜(AFM) 自组装 预混合 4-氨基安替比林 聚苯乙烯磺酸钠 阳离子聚电解质 生物催化活性 混合溶液 H2O2 阳离子化 对苯二酸 PDDA 米氏常数 显色反应 

分 类 号：TB43[一般工业技术]