Cloning and Expression of the Organophosphate Pesticide-Degradingα-βHydrolase Gene in Plasmid pMK-07 to Confer Cross-Resistance to Antibiotics

Abstract

Pesticide residual persistence in agriculture soil selectively increases the pesticide-degrading population and transfers the pesticide-degrading gene to other populations, leading to cross-resistance to a wide range of antibiotics. The enzymes that degrade pesticides can also catabolize the antibiotics by inducing changes in the gene or protein structure through induced mutations. The present work focuses on the pesticide-degrading bacteria isolated from an agricultural field that develop cross-resistance to antibiotics. This cross-resistance is developed through catabolic gene clusters present in an extrachromosomal plasmid. A larger plasmid (236.7 Kbp) isolated from Bacillus sp. was sequenced by next-generation sequencing, and important features such as α-β hydrolase, DNA topoisomerase, DNA polymerase III subunit beta, reverse transcriptase, plasmid replication rep X, recombination U, transposase, and S-formylglutathione hydrolase were found in this plasmid. Among these, the α-β hydrolase enzyme is known for the degradation of organophosphate pesticides. The cloning and expression of the α-β hydrolase gene imply nonspecific cleavage of antibiotics through a cross-resistance phenomenon in the host. The docking of α-β hydrolase with a spectrum of antibiotics showed a high G-score against chloramphenicol (−3.793), streptomycin (−2.865), cefotaxime (−5.885), ampicillin (−4.316), and tetracycline (−3.972). This study concludes that continuous exposure to pesticide residues may lead to the emergence of multidrug-resistant strains among the wild microbial flora.

Efficacy of soil‐borne Enterobacter sp. for carbofuran degradation: HPLC quantitation of degradation rate

“…It was well known that these special characteristics like degradation generally derived from the plasmid system of a certain bacteria. Some of the previously reported studies suggested the presence of such degrading genes in the plasmid [38, 39]. However, even after using two different techniques we could not identify the existence of any plasmid in the isolated bacteria [30].…”

Cultivating antimicrobial resistance: how intensive agriculture ploughs the way for antibiotic resistance

“…The organophosphate degradation genes ( opd ) have been identified in a plethora of soil-dwelling bacteria such as Flavobacterium , Pseudomonas , Bacillus and Agrobacterium [114–116]. However, enzymatic modelling studies by Rangasamy et al [117–119] have suggested that organophosphorus hydrolase from Geobacillus could dock with and hydrolyse streptomycin, ampicillin, chloramphenicol and cefotaxime. Furthermore, a plasmid-bound α–β hydrolase known to degrade organophosphate-degrading α–β hydrolases has been shown to reside on plasmids and provide resistance to a range of antibiotics [119].…”

Discovery of Novel Pentacyclic Triterpene Acid Amide Derivatives as Excellent Antimicrobial Agents Dependent on Generation of Reactive Oxygen Species

“…Of note, chemical pesticides are currently the most common and effective method for controlling plant bacterial diseases, providing an essential guarantee for global food production in recent decades [ 8 , 9 ]. However, the use of such traditional chemicals also brings some intractable problems, such as bacterial resistance [ 10 ], environmental pollution [ 11 ], ecological problems, and human safety concerns [ 12 ]. Therefore, an increasing need to develop agricultural bactericides that are both environmentally friendly and highly effective in controlling plant bacterial diseases is present.…”