Isolation of the <Emphasis Type="Italic">opdE</Emphasis> gene that encodes for a new hydrolase of <Emphasis Type="Italic">Enterobacter</Emphasis> sp. capable of degrading organophosphorus pesticides |
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Authors: | Concepción Chino-Flores Edgar Dantán-González Alejandra Vázquez-Ramos Raunel Tinoco-Valencia Rafael Díaz-Méndez Enrique Sánchez-Salinas Ma Luisa Castrejón-Godínez Fernando Ramos-Quintana Ma Laura Ortiz-Hernández |
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Institution: | 1.Laboratorio de Investigaciones Ambientales, Centro de Investigación en Biotecnología,Universidad Autónoma del Estado Morelos,Cuernavaca,Mexico;2.Instituto de Biotecnología,UNAM,Cuernavaca,Mexico;3.Centro de Ciencias Genómicas,UNAM,Cuernavaca,Mexico;4.Instituto Tecnológico de Estudios Superiores de Monterrey,Cuernavaca,Mexico |
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Abstract: | Microbial enzymes that can hydrolyze organophosphorus compounds have been isolated, identified and characterized from different
microbial species in order to use them in biodegradation of organophosphorus compounds. We isolated a bacterial strain Cons002
from an agricultural soil bacterial consortium, which can hydrolyze methyl-parathion (MP) and other organophosphate pesticides.
HPLC analysis showed that strain Cons002 is capable of degrading pesticides MP, parathion and phorate. Pulsed-field gel electrophoresis
and 16S rRNA amplification were performed for strain characterization and identification, respectively, showing that the strain
Cons002 is related to the genus Enterobacter sp. which has a single chromosome of 4.6 Mb and has no plasmids. Genomic library was constructed from DNA of Enterobacter sp. Cons002. A gene called opdE (Organophosphate Degradation from E
nterobacter) consists of 753 bp and encodes a protein of 25 kDa, which was isolated using activity methods. This gene opdE had no similarity to any genes reported to degrade organophosphates. When kanamycin-resistance cassette was placed in the
gene opdE, hydrolase activity was suppressed and Enterobacter sp. Cons002 had no growth with MP as a nutrients source. |
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