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 | |
刊名 | SENSORS AND ACTUATORS B-CHEMICAL |
2019-01-15 | |
卷号 | 279页码:483-489 |
关键词 | Carp Acetylcholinesterase (AChE) Yeast Cell surface display Organophosphate pesticide detection |
ISSN号 | 0925-4005 |
DOI | 10.1016/j.snb.2018.09.119 |
英文摘要 | 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. |
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关键词 | MICROBIAL BIOSENSOR ; SENSITIVE ACETYLCHOLINESTERASE ; NIPPOSTRONGYLUS-BRASILIENSIS ; GLUTAMATE-DEHYDROGENASE ; GLUCOSE-DEHYDROGENASE ; XYLOSE DEHYDROGENASE ; PICHIA-PASTORIS ; NERVE AGENTS ; HYDROLASE ; EXPRESSION |
WOS研究方向 | Chemistry ; Electrochemistry ; Instruments & Instrumentation |
语种 | 英语 |
出版者 | ELSEVIER SCIENCE SA |
WOS记录号 | WOS:000449373800060 |
资助机构 | National Natural Science Foundation of China ; Major Program of Shandong Province Natural Science Foundation |
内容类型 | 期刊论文 |
源URL | [http://ir.qibebt.ac.cn/handle/337004/11010] |
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 |
推荐引用方式 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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