一种基于新颖通用设计的用于L-苹果酸的双酶电极。

A bienzyme electrode for L-malate based on a novel and general design.

作者信息

Gajovic N, Warsinke A, Scheller F W

机构信息

Potsdam University, Institute of Biochemistry and Molecular Physiology, Berlin, Germany.

出版信息

J Biotechnol. 1998 Apr 15;61(2):129-33. doi: 10.1016/s0168-1656(98)00029-7.

DOI:10.1016/s0168-1656(98)00029-7

PMID:9654746

Abstract

The coimmobilization of a NAD(P) + -dependent dehydrogenase with salicylate hydroxylase (SHL, EC 1.14.13.1) in front of a Clark-electrode yields a flexible new design for dehydrogenase based biosensors. The feasibility of the approach has been tested with malic enzyme (MDH, EC 1.1.1.40) as the dehydrogenase, resulting in a novel L-malate sensor. It had substantial advantages over the biosensor approaches reported earlier: effective re-oxidation of NADPH by SHL yielded an extended linear range from 0.01 to 1.2 mmol 1(-1) L-malate and strongly reduced NADP+ -requirement (<0.025 mmol 1(-1)), while the working stability was increased to more than 30 days. The results obtained from six real samples showed a close correlation with the standard enzymatic method. The presented scheme with SHL and the Clark-electrode can be employed together with any NAD(P)+ -dependent dehydrogenase.

摘要

将一种NAD(P)+依赖性脱氢酶与水杨酸羟化酶（SHL，EC 1.14.13.1）共同固定在Clark电极前，可为基于脱氢酶的生物传感器带来一种灵活的新设计。该方法的可行性已通过以苹果酸酶（MDH，EC 1.1.1.40）作为脱氢酶进行测试，从而得到一种新型L-苹果酸传感器。它相对于早期报道的生物传感器方法具有显著优势：SHL对NADPH的有效再氧化使得L-苹果酸的线性范围从0.01扩展至1.2 mmol 1(-1)，并大幅降低了对NADP+的需求（<0.025 mmol 1(-1)），同时工作稳定性提高到超过30天。从六个实际样品中获得的结果与标准酶法显示出密切相关性。所提出的使用SHL和Clark电极的方案可与任何NAD(P)+依赖性脱氢酶一起使用。