QIBEBT-IR
Functional cell surface displaying of acetylcholinesterase for spectrophotometric sensing organophosphate pesticide
Liang, Bo1,2,3,4; Wang, Ge1,2,5; Yan, Lu1,2; Ren, Han6; Feng, Ruirui3,4; Xiong, Zhong5; Liu, Aihua1,2,5,6
2019-01-15
发表期刊SENSORS AND ACTUATORS B-CHEMICAL
ISSN0925-4005
卷号279页码:483-489
摘要The inhibition of acetylcholinesterase (AChE) activity based methods have been widely used for rapid detection of organophosphate (OP) pesticide residues. However, the sensitivity of the enzyme toward pesticides is not desirable and complex purification steps increase production cost. In this study, we optimized and synthesized the cDNA of carp AChE based on the preferred codon using Saccharomyces cerevisiae, which was ligated into expression vector pYD1. The AChE was successfully presented on yeast surface through a-agglutinin mediated display system. In shake-flask after induction with galactose in YPD medium for 24 h, this strain expressed AChE activity at a high level of 0.15 U/OD600/ml. The correct location of AChE was confirmed by immunofluorescence analysis. The whole cell catalyst was applied to visible spectrophotometry detection for OP pesticides based on inhibition rate. The inhibition rate was linear with paraoxon concentration in the range of 0.5 ng/ml-10 mu g/ml and parathion concentration in the range of 5 ng/ml-10 mu g/ml. The low detection limit was 0.136 ng/ml paraoxon and 3.72 ng/ml parathion (S/N = 3). The proposed method is applicable to analyzing OP pesticides in real samples. Therefore, the recombinant strain developed could be an efficient and affordable biomaterial for rapid and accurate detection of OP pesticide residues.
关键词Carp Acetylcholinesterase (AChE) Yeast Cell surface display Organophosphate pesticide detection
DOI10.1016/j.snb.2018.09.119
关键词[WOS]MICROBIAL BIOSENSOR ; SENSITIVE ACETYLCHOLINESTERASE ; NIPPOSTRONGYLUS-BRASILIENSIS ; GLUTAMATE-DEHYDROGENASE ; GLUCOSE-DEHYDROGENASE ; XYLOSE DEHYDROGENASE ; PICHIA-PASTORIS ; NERVE AGENTS ; HYDROLASE ; EXPRESSION
语种英语
资助项目National Natural Science Foundation of China[81673172] ; National Natural Science Foundation of China[21475144] ; National Natural Science Foundation of China[91227116] ; National Natural Science Foundation of China[21275152] ; Major Program of Shandong Province Natural Science Foundation[ZR2018ZC0125]
WOS研究方向Chemistry ; Electrochemistry ; Instruments & Instrumentation
项目资助者National Natural Science Foundation of China ; Major Program of Shandong Province Natural Science Foundation
WOS类目Chemistry, Analytical ; Electrochemistry ; Instruments & Instrumentation
WOS记录号WOS:000449373800060
出版者ELSEVIER SCIENCE SA
引用统计
文献类型期刊论文
条目标识符http://ir.qibebt.ac.cn/handle/337004/11011
专题中国科学院青岛生物能源与过程研究所
通讯作者Liu, Aihua
作者单位1.Qingdao Univ, Inst Biosensing, 308 Ningxia Rd, Qingdao 266071, Peoples R China
2.Qingdao Univ, Coll Life Sci, 308 Ningxia Rd, Qingdao 266071, Peoples R China
3.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, 189 Songling Rd, Qingdao 266101, Peoples R China
4.Chinese Acad Sci, Key Lab Bioenergy, 189 Songling Rd, Qingdao 266101, Peoples R China
5.Qingdao Univ, Coll Chem & Chem Engn, Qingdao 266071, Peoples R China
6.Qingdao Univ, Med Coll, Sch Pharm, Dept Drug Metab & Anal, Qingdao 266021, Peoples R China
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GB/T 7714
Liang, Bo,Wang, Ge,Yan, Lu,et al. Functional cell surface displaying of acetylcholinesterase for spectrophotometric sensing organophosphate pesticide[J]. SENSORS AND ACTUATORS B-CHEMICAL,2019,279:483-489.
APA Liang, Bo.,Wang, Ge.,Yan, Lu.,Ren, Han.,Feng, Ruirui.,...&Liu, Aihua.(2019).Functional cell surface displaying of acetylcholinesterase for spectrophotometric sensing organophosphate pesticide.SENSORS AND ACTUATORS B-CHEMICAL,279,483-489.
MLA Liang, Bo,et al."Functional cell surface displaying of acetylcholinesterase for spectrophotometric sensing organophosphate pesticide".SENSORS AND ACTUATORS B-CHEMICAL 279(2019):483-489.
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