Biofuel cell and phenolic biosensor based on acid-resistant laccase-glutaraldehyde functionalized chitosan-multiwalled carbon nanotubes nanocomposite film
Biofuel cell and phenolic biosensor based on acid-resistant laccase-glutaraldehyde functionalized chitosan-multiwalled carbon nanotubes nanocomposite film
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DOI：
10.1016/j.bios.2008.11.026
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发表时间：
2009-03-15
影响因子：
12.6
通讯作者：
Yao, Shouzhuo
中科院分区：
文献类型：
--
作者：
Tan, Yueming;Deng, Wenfang;Yao, Shouzhuo
为了从变色栓菌（Trametes versicolor）中获得在酸性水溶液中显示其最大酶活性的漆酶（Lac），用戊二醛（GA）化学修饰生物聚合物壳聚糖（G）以形成GA官能化CS（GAfCS），然后使其与Lac反应以形成在弱酸性溶液中稳定的Lac-GAfCS复合物（两步方案），如石英晶体微量天平和耐久性测试所证实的。在2，2 '-连氮双（3-乙基苯并噻唑啉-6-磺酸）二铵盐（ABTS）存在下，Lac-GAfCS-多壁碳纳米管（MWC-NTs）/玻碳（GC）电极对O2还原表现出良好的催化活性，并考察了固定化Lac对O2还原的pH依赖性酶活性。以Lac-GAfCS-MWCNTs/GC电极为生物阴极，葡萄糖氧化酶（GOx）-GAfCS-MWCNTs/GC电极为生物阳极，在pH 5.0的醋酸盐缓冲溶液中制备了葡萄糖/空气生物燃料电池。生物燃料电池输出的最大功率密度为9.6 μ W/cm（2），开路电池电压为0.19V，短路电流密度为114 μ A/cm（2），分别用电化学噪声（ECN）装置测量。在Britton-Robinson缓冲溶液（pH 3.0）中，该电极用于邻苯二酚的测定，线性范围为0.1-50 μ M，检测限为20 nM。与直接使用GA进行一锅Lac-GA-CS或Lac-GA交联到Doppler Lac相比，在所提出的两步方案中使用大分子GAfCS被证明对酶活性的危害较小，因此更适合于固定化酶以构建生物燃料电池和生物传感器。这项工作可能有助于利用流行的生物相容性CS作为耐酸膜基质的许多其他生物技术的应用，并建议两步交联协议的其他生物分子的高活性固定。(C)2008 Elsevier B. V.保留所有权利。
To immobilize laccase (Lac) from Trametes versicolor that shows its maximum enzymatic activity in acidic aqueous solutions, the biopolymer chitosan (G) was chemically modified with glutaraldehyde (GA) to form GA functionalized CS (GAfCS), which was then allowed to react with Lac to form a Lac-GAfCS composite that is robust in weakly acidic solutions (two-step protocol), as confirmed by quartz crystal microbalance and durability tests. The Lac-GAfCS-multiwalled carbon nanotubes (MWC-NTs)/glassy carbon (GC) electrode exhibited good catalytic activity towards 02 reduction in the presence of 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) diammonium salt (ABTS), and the pH-dependent enzymatic activity of the immobilized Lac towards 02 reduction was examined. A glucose/air biofuel cell was fabricated, with the Lac-GAfCS-MWCNTs/GC electrode as the biocathode and a glucose oxidase (GOx)-GAfCS-MWCNTs/GC electrode as the bioanode in a Nafion membrane-separated acetate buffer solution (pH 5.0). The biofuel cell output a maximum power density of 9.6 mu W/cm(2), an open-circuit cell voltage of 0.19V, and a short-circuit current density of 114 mu A/cm(2), respectively, as measured with an electrochemical noise (ECN) apparatus. Furthermore, the Lac-GAfCS-MWCNTs/GC electrode was applied to determine catechol in Britton-Robinson buffer solution (pH 3.0), with a linear range of 0.1-50 mu M and a limit of detection of 20 nM. In comparison with the direct use of GA for one-pot Lac-GA-CS or Lac-GA crosslinking to immobilize Lac, the use of macromolecular GAfCS in the proposed two-step protocol was proven to be less harmful to the enzymatic activity and thus more suitable for immobilizing the enzyme to construct the biofuel cell and biosensor. This work may be helpful for exploiting the popular biocompatible CS as an acid-resistant film matrix for many other biotechnology applications, and the proposed two-step crosslinking protocol is recommended for high-activity immobilization of other biomolecules. (C) 2008 Elsevier B.V. All rights reserved.