Carbamate pesticides, a new type of broad-spectrum pesticides for controlling pests, mites, and weeds, are developed to address the shortcomings of organochlorine and organophosphorus pesticides. Their widespread use and slow degradation have led to environmental pollution, causing damage to ecosystems and human health. Managing pesticide residues is a pressing issue in the current environmental protection. This study aims to investigate the expression of SyEst870, a member of the SGNH/GDSL hydrolase family in Sphingobium yanoikuyae, in a prokaryotic system and evaluate the ability of the recombinant protein to degrade carbamate pesticides. The prokaryotic expression vector pET-32a-SyEst870 was constructed and transformed into the Escherichia coli BL21 for heterologous expression. The purified protein was studied in terms of enzyme activity and effects of temperature, pH, and metal ions on the enzyme activity, with p-nitrophenol acetate as the substrate and based on the standard curve of p-nitrophenol. LC-MS (liquid chromatography-mass spectrometry) was employed to examine the degradation effects of SyEst870 on carbaryl, metolcarb, and isoprocarb. GC-MS (gas chromatography-mass spectrometry) was employed to detect the degradation products of SyEst870 for the three pesticides. The soluble protein SyEst870 was successfully obtained through the heterologous expression in Escherichia coli, which yielded an enzyme with the activity of 677.5 U after affinity chromatography. SyEst870 exhibited degradation rates of 82.34%, 84.43%, and 92.87% for carbaryl, metolcarb, and isoprocarb, respectively, at an initial concentration of 100 mg/L within 24 h at 30 ℃ and pH 7.0. The primary degradation products of carbaryl were identified as α-naphthol and methyl isocyanate. Metolcarb was mainly degraded into m-cresol and methyl isocyanate, and isoprocarb was mainly degraded into 2-isopropylphenol and methyl isocyanate. Compared with the half-life of carbamate pesticides in the natural environment, which ranges from a few days to several weeks, the recombinant protein SyEst870 can rapidly eliminate the residues of carbamate pesticides. This study lays a foundation for addressing pesticide residues in the environment and in fruits and vegetables.
Escherichia coli/metabolism* ; Sphingomonadaceae/genetics* ; Recombinant Proteins/metabolism* ; Biodegradation, Environmental ; Esterases/metabolism* ; Pesticides/isolation & purification* ; Carbamates/isolation & purification*

OBJECTIVE: To establish the solid phase extraction (SPE) with GC/MS technology for fish poisoning cases to determine five pesticides in fishpond. METHODS: By three solid phase extraction column including Oasis HLB cartridge, Bond Elut C18 and SampliQ C18, the recovery rate was compared to extract and purify five pesticides in fishpond. The effects of different kinds and dosages of eluents on extract rate were also reviewed. RESULTS Using Bond Elut C18 as solid phase extraction column and 3 mL benzene as eluent, the linear range of mass concentration of five pesticides in fishpond was 1-50 μg/mL, and the correlation coefficient was 0.996 2-0.999 6. The limit of detection was 3.4-26 μg/L and the recovery was 61.49%-102.48%. The relative standard deviations was less than or equal to 3.01%. CONCLU-SION: With high sensitivity, good accuracy and precision, SPE-GC/MS has simple and quick operation and less solvent. It can be applied to determination of five pesticides in fishpond.
Gas Chromatography-Mass Spectrometry/methods* ; Pesticides/isolation & purification* ; Solid Phase Extraction ; Solvents ; Water Pollutants, Chemical/isolation & purification*

A Triazophos-degrading strain, Klebsiella sp. E6, was identified by soil enrichment technology from the soil sampled from the vicinity of a factory manufacturing Triazophos (TAP). The nutrient requirement of the strain is simple. It can use TAP as the sole sources of carbon, nitrogen and phosphorus. Comparison of the degradation rates revealed that the strain degraded TAP most effectively when TAP was used as a sole nitrogen source. No inhibition effect occurred when TAP concentration was high as 1000 mg/L in the case of TAP was used as the sole nitrogen source. Analysis of the intermediates of TAP metabolism indicated that TAP is firstly hydrolyzed into 1-phenyl-3-hydroxy-1,2,4-triazole and O,O-diethyl phosphorothioic acid. 1-phenyl-3-hydroxy-1,2,4-triazole was further mineralized into inorganic compounds. A degradation pathway of TAP was proposed. The experiment results demonstrated that the strain has potential in biodegradation of TAP pollutions.
Biodegradation, Environmental ; Environmental Pollutants ; metabolism ; Klebsiella ; isolation & purification ; metabolism ; Organothiophosphates ; metabolism ; Pesticides ; metabolism ; Soil Microbiology ; Triazoles ; metabolism

The research progress on Chinese medicine plant resources with pesticide activities, the active components and their reaction mechanism as well as the application and prospect were reviewed in this paper. Some proposals on the exploitation of traditional Chinese medicine plant origin pesticide were given. It is suggested to found compounds with pesticide activities from heat clearing and toxic clearing medicinal plants.
Fungicides, Industrial ; isolation & purification ; pharmacology ; Fusarium ; drug effects ; Insecticides ; isolation & purification ; pharmacology ; Lectins ; isolation & purification ; pharmacology ; Pesticides ; isolation & purification ; pharmacology ; Plant Oils ; isolation & purification ; pharmacology ; Plant Viruses ; drug effects ; Plants, Medicinal ; chemistry