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Amperometric L-glutamate biosensor based on bacterial cell-surface displayed glutamate dehydrogenase
Liang, Bo1,2,3; Zhang, Shu1,2,4; Lang, Qiaolin1,2; Song, Jianxia4; Han, Lihui4; Liu, Aihua1,2,3
2015-07-16
Source PublicationANALYTICA CHIMICA ACTA
Volume884Issue:1Pages:83-89
Abstract

A novel L-glutamate biosensor was fabricated using bacteria surface-displayed glutamate dehydrogenase (Gldh-bacteria). Here the cofactor NADP(+)-specific dependent Gldh was expressed on the surface of Escherichia coli using N-terminal region of ice nucleation protein (INP) as the anchoring motif. The cell fractionation assay and SDS-PAGE analysis indicated that the majority of INP-Gldh fusion proteins were located on the surface of cells. The biosensor was fabricated by successively casting polyethyleneimine (PEI)-dispersed multi-walled carbon nanotubes (MWNTs), Gldh-bacteria and Nafion onto the glassy carbon electrode (Nafion/Gldh-bacteria/PEI-MWNTs/GCE). The MWNTs could not only significantly lower the oxidation overpotential towards NAPDH, which was the product of NADP(+) involving in the oxidation of glutamate by Gldh, but also enhanced the current response. Under the optimized experimental conditions, the current-time curve of the Nafion/Gldh-bacteria/PEI-MWNTs/GCE was performed at +0.52 V (vs. SCE) by amperometry varying glutamate concentration. The current response was linear with glutamate concentration in two ranges (10 mu M-1 mM and 2-10 mM). The low limit of detection was estimated to be 2 mM glutamate (S/N = 3). Moreover, the proposed biosensor is stable, specific, reproducible and simple, which can be applied to real samples detection. (C) 2015 Elsevier B.V. All rights reserved.

SubtypeArticle
KeywordBacterial Surface Display Electrochemical Microbial Biosensor Amperometric Detection L-glutamate Biosensor Glutamate Dehydrogenase Displayed Bacteria
WOS HeadingsScience & Technology ; Physical Sciences
DOI10.1016/j.aca.2015.05.012
WOS KeywordGLASSY-CARBON ELECTRODE ; MONOSODIUM L-GLUTAMATE ; OXIDASE BIOSENSOR ; CAPILLARY ELECTROPHORESIS ; ESCHERICHIA-COLI ; FOOD SAMPLES ; ACID ; IMMOBILIZATION ; SYSTEM ; SENSOR
Indexed BySCI
Language英语
WOS Research AreaChemistry
WOS SubjectChemistry, Analytical
WOS IDWOS:000356237000012
Citation statistics
Cited Times:22[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qibebt.ac.cn/handle/337004/6477
Collection生物传感技术团队(过去)
Affiliation1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuels, Lab Biosensing, Qingdao 266101, Peoples R China
2.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Energy Genet, Qingdao 266101, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Ocean Univ China, Key Lab Marine Chem Theory & Technol, Minist Educ, Qingdao 266100, Peoples R China
Recommended Citation
GB/T 7714
Liang, Bo,Zhang, Shu,Lang, Qiaolin,et al. Amperometric L-glutamate biosensor based on bacterial cell-surface displayed glutamate dehydrogenase[J]. ANALYTICA CHIMICA ACTA,2015,884(1):83-89.
APA Liang, Bo,Zhang, Shu,Lang, Qiaolin,Song, Jianxia,Han, Lihui,&Liu, Aihua.(2015).Amperometric L-glutamate biosensor based on bacterial cell-surface displayed glutamate dehydrogenase.ANALYTICA CHIMICA ACTA,884(1),83-89.
MLA Liang, Bo,et al."Amperometric L-glutamate biosensor based on bacterial cell-surface displayed glutamate dehydrogenase".ANALYTICA CHIMICA ACTA 884.1(2015):83-89.
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