Cell surface display of organophosphorus hydrolase for sensitive spectrophotometric detection of p-nitrophenol substituted organophosphates

doi:10.1016/j.enzmictec.2013.10.006

QIBEBT-IR > 生物传感技术团队（过去）

	Cell surface display of organophosphorus hydrolase for sensitive spectrophotometric detection of p-nitrophenol substituted organophosphates
	Tang, Xiangjiang1,2,3 ; Liang, Bo1,2 ; Yi, Tuyong 4; Manco, Giuseppe 5; Palchetti, Ilaria 6; Liu, Aihua 1,2,3
	2014-02-05
发表期刊	ENZYME AND MICROBIAL TECHNOLOGY
卷号	55 页码:107-112
摘要	Organophosphates COPS) widely exist in ecosystem as toxic substances, for which sensitive and rapid analytical methods are highly requested. In the present work, by using N-terminal of ice nucleation protein (INP) as anchoring motif, a genetically engineered Escherichia coli (E. coli) strain surface displayed mutant organophosphorus hydrolase (OPH) (S5) with improved enzyme activity was successfully constructed. The surface location of INP-OPH fusion was confirmed by SDS-PAGE analysis and enzyme activity assays. The OPH-displayed bacteria facilitate the hydrolysis of p-nitrophenol (PNP) substituted organophosphates to generate PNP, which can be detected spectrometrically at 410 nm. Over 90% of the recombinant protein present on the surface of microbes demonstrated enhanced enzyme activity and long-term stability. The OPH activity of whole cells was 2.16 U/OD600 using paraoxon as its substrate, which is the highest value reported so far. The optimal temperature for OPH activity was around 55 degrees C and suspended cultures retained almost 100% of its activity over a period of one month at room temperature, exhibiting the better stability than free OPH. The recombinant E. coli strain could be employed as a whole-cell biocatalyst for detecting PNP substituted OPs at wider ranges and lower detection limits. Specifically, the linear ranges of the calibration curves were 0.5-150 mu M paraoxon, 1-200 mu M parathion and 2.5-200 mu M methyl parathion, and limits of detection were 0.2 mu M, 0.4 mu M and 1 mu M for paraoxon, parathion and methyl parathion, respectively (S/N = 3). These results indicate that the engineered OPH strain is a promising multifunctional bacterium that could be used for further large-scale industrial and environmental applications. (C) 2013 Elsevier Inc. All rights reserved.
文章类型	Article
关键词	Bacterial Surface Display Ice Nucleation Protein Organophosphorus Hydrolase P-nitrophenol Spectrophotometric Detection Of Organophosphates
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
DOI	10.1016/j.enzmictec.2013.10.006
关键词[WOS]	RECOMBINANT ESCHERICHIA-COLI ; ICE-NUCLEATION PROTEIN ; AMPEROMETRIC MICROBIAL BIOSENSOR ; PSEUDOMONAS-PUTIDA JS444 ; NERVE AGENTS ; PARATHION HYDROLASE ; WHOLE-CELL ; ELECTROCHEMICAL BIOSENSOR ; SIMULTANEOUS DEGRADATION ; GLUCOSE-DEHYDROGENASE
收录类别	SCI
语种	英语
WOS研究方向	Biotechnology & Applied Microbiology
WOS类目	Biotechnology & Applied Microbiology
WOS记录号	WOS:000331156000015
引用统计
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/6173
专题	生物传感技术团队（过去）
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Lab Biosensing, Qingdao 266101, Peoples R China 2.Chinese Acad Sci, Key Lab Bioenergy, Qingdao 266101, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Hunan Agr Univ, Coll Plant Protect, Hunan Prov Key Lab Biol & Control Plant Pests, Changsha 410128, Hunan, Peoples R China 5.Italian Natl Res Council, Inst Prot Biochem, I-80131 Naples, Italy 6.Univ Florence, Dipartimento Chim, I-50019 Sesto Fiorentino, Italy
推荐引用方式 GB/T 7714	Tang, Xiangjiang,Liang, Bo,Yi, Tuyong,et al. Cell surface display of organophosphorus hydrolase for sensitive spectrophotometric detection of p-nitrophenol substituted organophosphates[J]. ENZYME AND MICROBIAL TECHNOLOGY,2014,55:107-112.
APA	Tang, Xiangjiang,Liang, Bo,Yi, Tuyong,Manco, Giuseppe,Palchetti, Ilaria,&Liu, Aihua.(2014).Cell surface display of organophosphorus hydrolase for sensitive spectrophotometric detection of p-nitrophenol substituted organophosphates.ENZYME AND MICROBIAL TECHNOLOGY,55,107-112.
MLA	Tang, Xiangjiang,et al."Cell surface display of organophosphorus hydrolase for sensitive spectrophotometric detection of p-nitrophenol substituted organophosphates".ENZYME AND MICROBIAL TECHNOLOGY 55(2014):107-112.