Isolation of the opdE gene that encodes for a new hydrolase of Enterobacter sp. capable of degrading organophosphorus pesticides

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 Enterobacter) 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.

Cloning of an organophosphorus hydrolase (opdD) gene of Lactobacillus sakei WCP904 isolated from chlorpyrifos-impregnated kimchi and hydrolysis activities of its gene product for organophosphorus pesticides

“…However, relative activity of the OpdD enzyme against the P-O bond insecticides was much higher than against P-S bond insecticides (Table 1), which is consistent with the previously reported OpdB enzymic character [11], while the opdE gene of Enterobacter sp. had shown only the breakdown ability of P-S bond of some organophosphate pesticides instead of P-O bond [22]. However, minor variations of relative substrate activities were observed for the OpdD enzyme versus those reported for the OpdB enzyme.…”

“…Phosphotriesterase from different microorganisms has been found to hydrolyze OP insecticides, specifically hydrolyze phosphoester bonds, such as P-O, P-E, P-NC, and P-S [22]. The OpdD enzyme hydrolyzed a range of OP insecticides containing P-O and P-S bonds ( Table 1), indicating that the recombinant OpdD has broad substrate specificity.…”

Chlorpyrifos: pollution and remediation

“…Organophosphate hydrolyzing enzymes are collectively known as aryldialkylphosphatase/ organophosphorus hydrolase/phosphotriesterase. Chlorpyrifos degrading microorganisms are capable of producing either of these enzymes such as organophosphorus hydrolase (OPH) , phosphotriesterase (PTE) methyl parathion hydrolase (MPH) (Chino-Flores et al, 2012), and organophosphorus acid anhydrolase (OPAA) (Cheng et al 1993;Theriot and Grunden 2011). These enzymes differ in Environ Chem Lett protein sequence, 3D structure, and catalytic mechanism, but they also share several common features.…”

Bioremediation of Soils Contaminated with Pesticides: Experiences in Mexico

“…2000 ;Van Hamme et al . 2003 ;Yeomans et al 2004 ;Haritash and Kaushik 2009 ;Chino-Flores et al 2011 ;Abdel-Razek et al 2013 ). Pesticides are transformed by metabolic reactions leading to changes in their chemical structure through diverse reactions, which give place to inorganic compounds such as CO 2 , H 2 O, halides, ammonium, and phosphates, among others.…”