合成生物学在农残检测领域的应用

doi:10.11949/0438-1157.20201327

化工学报 ›› 2021, Vol. 72 ›› Issue (5): 2413-2425.DOI: 10.11949/0438-1157.20201327

合成生物学在农残检测领域的应用

毛金竹^1,³(),肖淑玲^1,²,杨智淳^1,²,王孝宇^1,⁴,张诗^1,⁵,陈俊宏^1,²,谢佶晟^1,²,陈福德^1,²,黄子诺^1,²,冯天宇^1,²,张瑷珲^1,^2,⁶(),方柏山^1,^2,^6,⁷()

^1.厦门大学遗传工程机器设计创新实践平台，福建厦门 361005
^2.厦门大学化学化工学院，福建厦门 361005
^3.厦门大学药学院，福建厦门 361005
^4.厦门大学材料学院，福建厦门 361005
^5.厦门大学公共卫生学院，福建厦门 361005
^6.厦门市合成生物技术重点实验室，福建厦门 361005
^7.福建省化学生物学重点实验室，福建厦门 361005

收稿日期:2020-09-17 修回日期:2020-12-23 出版日期:2021-05-05 发布日期:2021-05-05
通讯作者: 张瑷珲,方柏山
作者简介:毛金竹（1999—），女，硕士研究生，maojinzhunico@163.com

Application of synthetic biology in pesticides residues detection

MAO Jinzhu^1,³(),XIAO Shuling^1,²,YANG Zhichun^1,²,WANG Xiaoyu^1,⁴,ZHANG Shi^1,⁵,CHEN Junhong^1,²,XIE Jisheng^1,²,CHEN Fude^1,²,HUANG Zinuo^1,²,FENG Tianyu^1,²,ZHANG Aihui^1,^2,⁶(),FANG Baishan^1,^2,^6,⁷()

^1.Innovative Practice Platform for Genetic Engineering Machine Design, Xiamen University, Xiamen 361005, Fujian, China
^2.College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, Fujian, China
^3.School of Pharmaceutical Science, Xiamen University, Xiamen 361005, Fujian, China
^4.College of Materials, Xiamen University, Xiamen 361005, Fujian, China
^5.School of Public Health, Xiamen University, Xiamen 361005, Fujian, China
^6.The Key Laboratory for Synthetic Biotechnology of Xiamen City, Xiamen 361005, Fujian, China
^7.The Key Laboratory for Chemical Biology of Fujian Province, Xiamen 361005, Fujian, China

Received:2020-09-17 Revised:2020-12-23 Online:2021-05-05 Published:2021-05-05
Contact: ZHANG Aihui,FANG Baishan

摘要/Abstract

摘要：

近年来，合成生物学在多个领域崭露头角，在农残检测中也发挥着越来越重要的作用。基于合成生物学模块化和工程化指导思想，各种基因部件的多样化组合为农残检测提供更多方案。简便、耐用、低成本、原位检测等特点也使其较传统检测手段具有更强的竞争力。但与此同时，合成生物学在农残检测中的应用也受到复杂检测环境和生物安全性等问题的影响。结合目前合成生物学在有机氯、有机磷、拟除虫菊酯和氨基甲酸酯类农药检测中的应用与创新实例，归纳合成生物学在农残检测中应用的原理，分析并探讨合成生物学技术未来在农残检测中的发展潜力与应用前景。

关键词: 合成生物学, 农残检测, 生物反应器, 酶, 生物技术

Abstract:

In recent years, synthetic biology has emerged in many fields, and it has also played an increasingly important role in pesticide residue detection. Through synthetic biology, a variety of gene assemblies provide more potential for detection. The characteristics of simplicity, rapidity, durability, and low cost grow on its competence comparing with traditional detection methods. The support of miniaturized instruments also enhances its application value in situ detection of pesticide residues. But at the same time, the application of synthetic biology in pesticide residue detection is also affected by the complex detection environment and biological safety issues. This review mainly introduces the application and innovation of synthetic biology in the detection of organochlorine pesticides, organophosphorus pesticides, pyrethroid pesticides, and carbamate pesticides. Analyze its advantages and shortages and discuss its future development potential.

Key words: synthetic biology, pesticide residue detection, bioreactor, enzyme, biotechnology

中图分类号:

Q 812

毛金竹, 肖淑玲, 杨智淳, 王孝宇, 张诗, 陈俊宏, 谢佶晟, 陈福德, 黄子诺, 冯天宇, 张瑷珲, 方柏山. 合成生物学在农残检测领域的应用[J]. 化工学报, 2021, 72(5): 2413-2425.

MAO Jinzhu, XIAO Shuling, YANG Zhichun, WANG Xiaoyu, ZHANG Shi, CHEN Junhong, XIE Jisheng, CHEN Fude, HUANG Zinuo, FENG Tianyu, ZHANG Aihui, FANG Baishan. Application of synthetic biology in pesticides residues detection[J]. CIESC Journal, 2021, 72(5): 2413-2425.

图/表 7

图1 世界农药使用量情况及其危害[1]

Fig.1 World pesticide use and its hazards [1]

图2 利用scFv-AP进行竞争性酶联免疫吸附测定（ELISA）[32]MAb—单克隆抗体；OVA—鸡卵白蛋白，半抗原偶联载体蛋白；VL—抗体轻链可变区；VH—抗体重链可变区；CL—抗体轻链恒定区；CH1，CH2，CH3—抗体重链恒定区

Fig.2 The competitive enzyme-linked immunosorbent assay (ELISA) was performed using scFv-AP[32]

图3 基于AChE的高灵敏度荧光生物传感器[46]

Fig.3 High sensitivity fluorescent biosensor based on AChE [46]

表1 合成生物学在农残检测中的应用

Table 1 Application of synthetic biology in pesticide residue detection

化合物	关键基因元件	底盘生物	输出信号	检测限（LOD）	文献
林丹（γ-HCH）	linA2	E. coli	电导率	2×10^-12	[48]
阿特拉津	atrABC atzR、atzDEF	E. coli SM004	生物发光（LuxCDABE）	1.08 μmol/L	[49]
对氧磷	opd	E. coli DH5α	pH	1 μmol/L	[21]
有机磷	opd	E. coli	pH	2 μmol/L	[50]
有机磷	opd、mpd	E. coli XL1-Blue	荧光（GFP）	2 μmol/L	[22]
对氧磷	opd	S. cerevisiae MT8-1	荧光（EGFP）	—	[51]
对氧磷和对硫磷、蝇毒磷	opd	E. coli	光纤	对氧磷和对硫磷（3 μmol/L）蝇毒磷（5 μmol/L）	[52]
有机磷	opd	E. coli BL21	吸光度	对氧磷（0.2 μmol/L）对硫磷（0.4 μmol/L）甲基对硫磷（1 μmol/L）	[53]
有机磷	opd	Pseudomonas putida JS444	电流	对氧磷（0.28×10^-9）甲基对硫磷（0.26×10^-9）对硫磷（0.29×10^-9）	[54]
有机磷	opd	Moraxella sp.	电流	对氧磷（0.2 μmol/L）甲基对硫磷（1 μmol/L）	[55]
毒死蜱	chpR、chpA-atsBA	E. coli	生物发光	25 nmol/L	[56]
乙基对氧磷	opd、pnpR-pnpC	E. coli XL1-Blue E. coli DH5α	颜色	1 nmol/L	[57]
3-苯氧基苯甲酸	VHH、amilCP	E. coli	沉淀	3 ng/ml	[58]
呋喃丹	H5 DNA	E. coli HB101	荧光（EGFP）	1 ng/L	[59]

图4 农药分子特异性激活转录调节因子

Fig.4 Pesticide specific activation of transcription regulators

图5 pH玻璃电极检测有机磷[50]

Fig.5 Detection of organophosphorus by pH glass electrode [50]

图6 表面展示水解酶及绿色荧光蛋白[22,51](a) 荧光蛋白与水解酶融合; (b) 荧光蛋白与水解酶共同表面展示OPs/OP—有机磷；MPH—甲基对硫磷水解酶；OPH—有机磷水解酶；GFP/EGFP—绿色荧光蛋白；OM—细胞外膜；INP N/C—冰核蛋白；LPP/OmpA—大肠杆菌表面展示系统

Fig.6 Surface display of hydrolase and green fluorescent protein [22,51]

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合成生物学在农残检测领域的应用

Application of synthetic biology in pesticides residues detection

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