Zeng Xiandong, Wei Wanzhi, Li Xuefang, Zeng Jinxiang, Wu Ling
State Key Laboratory of Chemo/Biosensoring and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Hunan, Changsha 410082, P. R. China.
Bioelectrochemistry. 2007 Nov;71(2):135-41. doi: 10.1016/j.bioelechem.2007.02.006. Epub 2007 Mar 1.
Semi-interpenetrating polymer network (semi-IPN) hydrogel based on polyacrylamide (PAM) and chitosan was prepared to immobilize redox protein hemoglobin (Hb). The Hb-PAM-chitosan hydrogel film obtained has been investigated by scanning electron microscopy (SEM) and UV-VIS spectroscopy. UV-VIS spectroscopy showed that Hb kept its secondary structure similar to its native state in the Hb-PAM-chitosan hydrogel film. Cyclic voltammogram of Hb-PAM-chitosan film-modified glass carbon (GC) electrode showed a pair of well-defined and quasi-reversible redox peaks for Hb Fe(III)/Fe(II), indicating that direct electron transfer between Hb and GC electrode occurred. The electron-transfer rate constant was about 5.51 s(-1) in pH 7.0 buffers, and the formal potential (E degrees ') was -0.324 V (vs. SCE). The dependence of E degrees ' on solution pH indicated that one-proton transfer was coupled to each electron transfer in the direct electron-transfer reaction. Additionally, Hb in the semi-IPN hydrogel film retained its bioactivity and showed excellent electrocatalytic activity toward H(2)O(2). The electrocatalytic current values were linear with increasing concentration of H(2)O(2) in a wide range of 5-420 microM. The unique semi-IPN hydrogel would have wide potential applications in direct electrochemistry, biosensors and biocatalysis.
制备了基于聚丙烯酰胺(PAM)和壳聚糖的半互穿聚合物网络(semi-IPN)水凝胶,用于固定氧化还原蛋白血红蛋白(Hb)。通过扫描电子显微镜(SEM)和紫外可见光谱对所得的Hb-PAM-壳聚糖水凝胶膜进行了研究。紫外可见光谱表明,Hb在Hb-PAM-壳聚糖水凝胶膜中保持了与其天然状态相似的二级结构。Hb-PAM-壳聚糖膜修饰玻碳(GC)电极的循环伏安图显示出一对明确且准可逆的Hb Fe(III)/Fe(II)氧化还原峰,表明Hb与GC电极之间发生了直接电子转移。在pH 7.0缓冲溶液中,电子转移速率常数约为5.51 s(-1),形式电位(E°')为 -0.324 V(相对于饱和甘汞电极,SCE)。E°'对溶液pH的依赖性表明,在直接电子转移反应中,每一次电子转移伴随着一个质子转移。此外,半互穿聚合物网络水凝胶膜中的Hb保留了其生物活性,并对H₂O₂表现出优异的电催化活性。在5 - 420 μM的宽范围内,电催化电流值与H₂O₂浓度的增加呈线性关系。这种独特的半互穿聚合物网络水凝胶在直接电化学、生物传感器和生物催化领域具有广泛的潜在应用。