title: Construction of a genetically engineered microorganism that simultaneously degrades organochlorine and organophosphate pesticides.
abstract: Field contamination with pesticide mixtures of organophosphates (OPs) and organochlorines (OCs) is becoming global issues to be solved urgently. The strategy of utilizing engineered microorganisms that have an ability to simultaneously degrade OPs and OCs has increasingly received great interest. In this work, an OP degradation gene (mpd) and an OC degradation gene (linA) were simultaneously introduced into Escherichia coli by using two compatible plasmids, resulting in strains with both OP degradation and OC degradation capabilities. To overcome the potential substrate uptake limitation, MPH was displayed on the cell surface of Escherichia coli using the N- and C-terminal domains of ice nucleation protein (INPNC) as an anchoring motif. The surface localization of INPNC-MPH was verified by cell fractionation, Western blot, proteinase accessibility, and immunofluorescence microscopy. Furthermore, both LinA and green fluorescent protein (GFP) were functionally co-expressed in the MPH-displaying Escherichia coli. The engineered Escherichia coli degraded OPs as well as OCs rapidly, and it can be easily monitored by GFP fluorescence.
authors.affiliation: Tai Shan University, Taian 271021, China.
authors.forename: Jijian
authors.initials: J
authors.lastname: Yang
authors.forename: Ruihua
authors.initials: R
authors.lastname: Liu
authors.forename: Wenli
authors.initials: W
authors.lastname: Song
authors.forename: Yao
authors.initials: Y
authors.lastname: Yang
authors.forename: Feng
authors.initials: F
authors.lastname: Cui
authors.forename: Chuanling
authors.initials: C
authors.lastname: Qiao
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001673:Biodegradation, Environmental; D002458:Cell Fractionation; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics; D006843:Hydrocarbons, Chlorinated / Q000378:metabolism*; D007306:Insecticides / Q000378:metabolism; D008190:Lyases / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D030781:Organisms, Genetically Modified; D010755:Organophosphates / Q000378:metabolism*; D010575:Pesticides / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D006843:Hydrocarbons, Chlorinated; D007306:Insecticides; D010755:Organophosphates; D010575:Pesticides; D011993:Recombinant Fusion Proteins; C063113:ice nucleation protein; D049452:Green Fluorescent Proteins; D010744:Phosphoric Monoester Hydrolases; D008190:Lyases; C071445:linA protein, Pseudomonas paucimobilis