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Highly sensitive microbial biosensor based on recombinant Escherichia coli overexpressing catechol 2,3-dioxygenase for reliable detection of catechol
- Source :
- Biosensors and Bioelectronics. 126:51-58
- Publication Year :
- 2019
- Publisher :
- Elsevier BV, 2019.
-
Abstract
- 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. coli 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. coli 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. coli BL21 over-expressed C23O. Based on the synergistic effect of C23O and NPG, the E. coli BL21-C23O/NPG/GCE bioelectrode showed a good linear response for catechol detection ranging from 1 μM to 500 μM with a high sensitivity of 332.24 μA mM−1 cm−2 and a low detection limit of 0.24 μM. Besides, the E. coli 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.
- Subjects :
- Catechols
Biomedical Engineering
Biophysics
Biosensing Techniques
02 engineering and technology
medicine.disease_cause
01 natural sciences
Catechol 2,3-Dioxygenase
Sphingobium
law.invention
chemistry.chemical_compound
Limit of Detection
Dioxygenase
law
Escherichia coli
Electrochemistry
medicine
Electrodes
Detection limit
chemistry.chemical_classification
Catechol
Chromatography
biology
010401 analytical chemistry
General Medicine
021001 nanoscience & nanotechnology
biology.organism_classification
Recombinant Proteins
Up-Regulation
0104 chemical sciences
Sphingomonadaceae
Enzyme
chemistry
Recombinant DNA
bacteria
0210 nano-technology
Biosensor
Biotechnology
Subjects
Details
- ISSN :
- 09565663
- Volume :
- 126
- Database :
- OpenAIRE
- Journal :
- Biosensors and Bioelectronics
- Accession number :
- edsair.doi.dedup.....f63079d86f25494df72c780e03c0e132