Highly sensitive microbial biosensor based on recombinant Escherichia coli overexpressing catechol 2,3-dioxygenase for reliable detection of catechol
Highly sensitive microbial biosensor based on recombinant Escherichia coli overexpressing catechol 2,3-dioxygenase for reliable detection of catechol
复制标题
基于重组大肠杆菌过表达儿茶酚 2,3-双加氧酶的高灵敏度微生物生物传感器，用于可靠检测儿茶酚
DOI：
10.1016/j.bios.2018.10.050
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发表时间：
2019-02-01
影响因子：
12.6
通讯作者：
Wang, Xia
中科院分区：
文献类型：
--
作者：
Liu, Zhuang;Zhang, Yachao;Wang, Xia
本研究建立了一种高灵敏度的基于全细胞的邻苯二酚电化学生物传感器。克隆了矢野氏鞘氨醇菌（Sphingobium yanoikuyae）XLDN 2 -5的carE基因，并在大肠杆菌中高效表达。BL21（E. coli BL 21）。与矢野氏鞘氨醇杆菌XLDN 2 -5相比，重组E. coil BL 21过表达的C23 O对邻苯二酚具有较高的催化活性。与粗酶相比，全细胞为C23 O提供了更好的环境，使其保持了催化活性和稳定性。重组E. coil BL 21过表达C23 O，使其成为制备微生物传感器的理想生物识别元件。此外，还选择了具有独特结构和功能特性的纳米多孔金（NPG）作为载体固定化重组大肠杆菌。conBL 21过表达C23 O。基于C23 O和NPG的协同效应，E.结果表明，BL 21-C23 O/NPG/GCE生物电极对邻苯二酚的线性范围为1 μ M ~ 500 μ M，灵敏度为332.24 μ A·mM ~（-1）·cm ~（-2），检测限为0.24 μ M。此外，E.结果表明，BL 21-C23 O/NPG/GCE螺旋电极抗干扰能力强，稳定性好。对于废水样品中邻苯二酚的检测，采用E. coliBL 21-C23 O/NPG/GCE生物电极与废水样品中添加的标准浓度符合良好，使E. coli BL 21-C23 O/NPG/GCE生物电极是一种理想的邻苯二酚检测工具。
A highly sensitive whole cell based electrochemical biosensor was developed for catechol detection in this study. The carE gene of Sphingobium yanoikuyae XLDN2-5 encoding catechol 2,3-dioxygenase (C23O), a key enzyme in the biodegradation of aromatic compound, was cloned and over-expressed in Escherichia coli. BL21 (E. coli BL21). Compared to Sphingobium yanoikuyae XLDN2-5, the recombinant E. coil BL21 over-expressed C23O exhibited higher catalytic activity towards catechol. Moreover, the whole cells provided a better environment for C23O to maintain its catalytic activity and stability compared with crude enzyme. The distinctive features of the recombinant E. coil BL21 over-expressed C23O made it an ideal bio-recognition element for the fabrication of a microbial biosensor. Additionally, nanoporous gold (NPG) with unique properties of structure and function was selected as a support to immobilized the recombinant E. con BL21 over-expressed C23O. Based on the synergistic effect of C23O and NPG, the E. coil BL21-C23O/NPG/GCE bioelectrode showed a good linear response for catechol detection ranging from 1 mu M to 500 mu M with a high sensitivity of 332.24 mu A mM(-1) cm(-2) and a low detection limit of 0.24 mu M. Besides, the E. coil BL21-C23O/NPG/GCE bioelectrode exhibited strong anti-interference and good stability. For the detection of catechol in wastewater samples, the concentrations detected by the E. coli BL21-C23O/NPG/GCE bioelectrode were in good agreement with the standard concentrations that added in the wastewater samples, which make the E. coli BL21-C23O/NPG/GCE bioelectrode an ideal tool for reliable catechol detection.