title: Construction of genetically engineered bacteria that degrades organophosphorus pesticide residues and can be easily detected by the fluorescence.
abstract: Organophosphorus compounds (OPs) are widely used in agriculture and industry and there is increased concern about their toxicological effects in the environment. Bioremediation can offer an efficient and cost-effective option for the removal of OPs. Herein, we describe the construction of a genetically engineered microorganism (GEM) that can degrade OPs and be directly detected and monitored in the environment using an enhanced green fluorescent protein (EGFP) fusion strategy. The coding regions of EGFP, a reporter protein that can fluoresce by itself, and organophosphorus hydrolase (OPH), which has a broad substrate specificity and is able to hydrolyse a number of organophosphorus pesticides, were cloned into the expression vector pET-28b. The fusion protein of EGFP-OPH was expressed in E. coli BL21 (DE3) and the protein expression reached the highest level at 11 h after isopropyl beta-D-thiogalactopyranoside induction. The fluorescence of the GEM was detected by fluorescence spectrophotometry and microscopy, and its ability to degrade OPs was determined by OPH activity assay. Those GEM that express the fusion protein (EGFP and OPH) exhibited strong fluorescence intensity and also potent hydrolase activity, which could be used to degrade organophosphorus pesticide residues in the environment and can also be directly monitored by fluorescence.
authors.forename: Qin
authors.initials: Q
authors.lastname: Li
authors.forename: Pan
authors.initials: P
authors.lastname: Wang
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Yi-Jun
authors.initials: YJ
authors.lastname: Wu
mesh_terms: D000383:Agriculture; D043303:Aryldialkylphosphatase / Q000737:chemistry; D001419:Bacteria / Q000235:genetics* / Q000378:metabolism; D001673:Biodegradation, Environmental; D002384:Catalysis; D004784:Environmental Monitoring / Q000379:methods; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006867:Hydrolases / Q000737:chemistry; D007221:Industry; D008856:Microscopy, Fluorescence; D009943:Organophosphorus Compounds / Q000378:metabolism*; D010278:Parathion / Q000737:chemistry; D010573:Pesticide Residues / Q000032:analysis* / Q000378:metabolism*; D010575:Pesticides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013862:Thiogalactosides / Q000737:chemistry; D013997:Time Factors
chemical_list: D009943:Organophosphorus Compounds; D010573:Pesticide Residues; D010575:Pesticides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013862:Thiogalactosides; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D010278:Parathion; D006867:Hydrolases; D043303:Aryldialkylphosphatase