title: Genetic engineering of Stenotrophomonas strain YC-1 to possess a broader substrate range for organophosphates.
abstract: In this work, Stenotrophomonas sp. strain YC-1, a native soil bacterium that produces methyl parathion hydrolase (MPH), was genetically engineered to possess a broader substrate range for organophosphates (OPs). A surface anchor system derived from the truncated ice nucleation protein (INPNC) from Pseudomonas syringae was used to target organophosphorus hydrolase (OPH) onto the cell surface of strain YC-1, reducing the potential substrate uptake limitation. The surface localization of INPNC-OPH was verified by cell fractionation, Western blot, proteinase accessibility, and immunofluorescence microscopy. No growth inhibition was observed for the engineered strain, and suspended cultures retained almost 100% activity over a period of 2 weeks. Concomitant expression of OPH in strain YC-1 resulted in a recombinant strain capable of simultaneously degrading diethyl and dimethyl OPs. A mixture of six OP pesticides (0.2 mM each) could be degraded completely within 5 h. The broader substrate specificity in combination with the rapid degradation rate makes this engineered strain a promising candidate for in situ remediation of OP-contaminated sites.
authors.affiliation: State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Yang
authors.forename: Cunjiang
authors.initials: C
authors.lastname: Song
authors.forename: Ashok
authors.initials: A
authors.lastname: Mulchandani
authors.forename: Chuanling
authors.initials: C
authors.lastname: Qiao
mesh_terms: D043303:Aryldialkylphosphatase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001673:Biodegradation, Environmental; D005818:Genetic Engineering; D010755:Organophosphates / Q000378:metabolism*; D010575:Pesticides / Q000378:metabolism*; D020588:Stenotrophomonas / Q000737:chemistry / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D010755:Organophosphates; D010575:Pesticides; D043303:Aryldialkylphosphatase