生物传感细胞ADP1_pWHlux在水环境急性毒性检测中的应用

针对天然发光菌和以模式微生物为宿主构建的生物传感细胞在急性毒性检测应用中对测试条件要求苛刻等适用性问题,本研究将1株基因工程构建的生物传感细胞不动杆菌ADP1_p WHlux应用于急性毒性检测,建立检测方法,考察其灵敏度及检测范围.结果表明,ADP1_p WHlux发光受急性毒物的抑制,毒物剂量与发光抑制存在剂量效应关系.在4 mg·L-1Hg Cl2诱导下仅5 min可作出响应,暴露30~60 min后可以给出较为准确的结果.对Hg Cl2的检出限可达0.04 mg·L-1.对我国生活饮用水卫生标准的指标中Be2+、Ba2+、Cu2+、Ni2+检出效果明显,对Be2+、Ba2+、Cu2+的检测范围均在0.025~250 mg·L-1,对Ni2+的检测范围在0.002 5~250 mg·L-1,对Pb2+、Br O-3、Cl O-2的检出限均在0.002 5 mg·L-1,对Cl O-3检出限为0.025mg·L-1.采用ADP1_p WHlux生物传感细胞检测方法对北京市清河水环境急性毒性进行评价,表明ADP1_p WHlux生物传感细胞检测方法可用于污染水样检测.

Application of Whole-cell Biosensor ADP1_pWHlux for Acute Toxicity Detection in Water Environment

A whole-cell biosensor acinetobacter ADP1_pWHlux was constructed by genetic engineering for detecting acute toxicity, so as to overcome the harsh application conditions when detecting acute toxicity using natural luminescent bacteria or whole-cell biosensor constructed by model microorganisms as the host cell,.Detection methods, detection sensitivity and detection range of acinetobacter ADP1_pWHlux were studied.The results showed that the luminescence of ADP1_pWHlux was inhibited by acute poison, poison dose and inhibition of luminescence exhibit dose-response relationship.ADP1_pWHlux was respond to 4 mg·L^-1 HgCl₂ within 5 min.The detection limit for HgCl₂ was 0.04 mg·L^-1.The detectable effects for indicators of Be²⁺, Ba²⁺, Cu²⁺, Ni²⁺ in standards for drinking water quality were obvious.The detection range of Be²⁺, Ba²⁺, Cu²⁺ were 0.025-250 mg·L^-1, the detection range of Ni²⁺ was 0.0025-250 mg·L^-1, the detection limit of Pb²⁺, BrO₃^-, ClO₂^- were 0.0025 mg·L^-1, the detection limit of ClO₃^- was 0.025 mg·L^-1.The whole-cell biosensor ADP1_pWHlux detection method has been applied to evaluate acute toxicity in water environment of Qinghe river in Beijing, indicating the established method can be used to detect contaminated water samples.