title: Construction of a green fluorescent protein (GFP)-marked multifunctional pesticide-degrading bacterium for simultaneous degradation of organophosphates and γ-hexachlorocyclohexane.
abstract: An autofluorescent whole-cell biocatalyst capable of simultaneously degrading organophosphates (OPs) and γ-hexachlorocyclohexane (γ-HCH) was constructed by display of organophosphorus hydrolase (OPH) and green fluorescent protein (GFP) fusion on the cell surface of a γ-HCH-degrading  Sphingobium japonicum  UT26 using the truncated ice nucleation protein (INPNC) as an anchoring motif. The surface localization of INPNC-OPH-GFP fusion was verified by cell fractionation, Western blot, proteinase accessibility, and immunofluorescence microscopy. Surface display of macromolecular OPH-GFP fusion (63 kDa) neither inhibits cell growth nor affects cell viability. In sterile and nonsterile soil samples, a mixture of parathion (100 mg kg(-1)) and γ-HCH (10 mg kg(-1)) could be degraded completely within 15 days when inoculated with the engineered UT26, and the strain could be easily monitored by fluorescence during bioremediation. These results indicate that the engineered UT26 is a promising multifunctional bacterium that could be used for the bioremediation of environments contaminated with multiple pesticides.
authors: Key Laboratory of Molecular Microbiology and Technology for Ministry of Education, Nankai University, Tianjin 300071, China.
authors: Chao
authors: C
authors: Yang
authors: Ruihua
authors: R
authors: Liu
authors: Yulan
authors: Y
authors: Yuan
authors: Jianli
authors: J
authors: Liu
authors: Xiangyu
authors: X
authors: Cao
authors: Chuanling
authors: C
authors: Qiao
authors: Cunjiang
authors: C
authors: Song
mesh_terms: D043303:Aryldialkylphosphatase / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D001556:Hexachlorocyclohexane / Q000378:metabolism*; D010755:Organophosphates / Q000378:metabolism*; D010575:Pesticides / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D042301:Sphingomonadaceae / Q000235:genetics / Q000378:metabolism*
chemical_list: D010755:Organophosphates; D010575:Pesticides; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D001556:Hexachlorocyclohexane; D043303:Aryldialkylphosphatase