Plasmid-Mediated Biodegradation of Chlorpyrifos and Analysis of Its Metabolic By-Products
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Plasmid‑Mediated Biodegradation of Chlorpyrifos and Analysis of Its Metabolic By‑Products Elizabeth M. John1 · Edna M. Varghese1 · Jisha M. Shaike1 Received: 30 April 2019 / Accepted: 6 July 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Organophosphate pesticide persistence is an emerging menace to the environment and despite this fact, its use has been enhanced due to its high efficiency. Bioremediation using microorganisms would be the only means by which these hazardous compounds could be wiped out without disturbing the environmental harmony. The current work studied the molecular mechanism of degradation of Chlorpyrifos (CP) by a bacterial consortium C5 comprising of three soil isolates Staphylococcus warneri (CPI 2), Pseudomonas putida (CPI 9) and Stenotrophomonas maltophilia (CPI 15), which unveiled that the property is plasmid borne. All the isolates were found to possess a 4 kb plasmid which could be cured only by using sodium azide. The Escherichia coli JM109 cells when transformed individually with the plasmid of the isolates showed CP degradation in mineral salts medium (MSM) that contained CP as the sole carbon source. The degradative enzyme organophosphorus hydrolase (~ 60 KDa) of the isolates was extracted and purified to 31.85, 26 and 37.74 fold, respectively. The possible metabolic by-products of CP degradation by the consortium C5, were also analysed. The LC-Q-Tof MS analysis revealed the presence of the major metabolite 3, 5, 6 -trichloropyridine (TCP) with the formation of chlorpyrifos oxon as the intermediate. The isolates also showed trichloropyridine degradation (> 80%) individually in MSM-TCP medium proving its efficiency to remediate both CP and TCP.
Introduction Bioremediation as an effective biotechnological approach has received increased attention to clean up environments polluted with Chlorpyrifos (CP), a commonly used organophosphate pesticide (OP), that persists in the environment for long time. However, the use of microorganisms for bioremediation requires an understanding of all physiological, microbiological, ecological, biochemical and molecular aspects involved in pollutant transformation [15, 18]. Several studies have given special emphasis on the role of catabolic genes in pesticide degradation and application of recombinant DNA technology to develop ‘superbugs’ that can simultaneously degrade several xenobiotics [21]. Currently, the
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00284-020-02115-y) contains supplementary material, which is available to authorized users. * Jisha M. Shaike [email protected] 1
School of Biosciences, Mahatma Gandhi University, Kottayam, Kerala 686560, India
opd (organophosphate degradation) and mpd (methyl parathion hydrolase) genes encoding the enzyme system involved in degradation of certain OPs have been identified and some of these catabolic genes have been shown to hydrolyze CP in microbes. Most of the opd genes were found to be plasmid based [31
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