Mineralization of Paraoxon and Its Use as a Sole C and P Source by a Rationally Designed Catabolic Pathway in Pseudomonas putida
- 1 October 2006
- journal article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 72 (10), 6699-6706
- https://doi.org/10.1128/aem.00907-06
Abstract
Organophosphate compounds, which are widely used as pesticides and chemical warfare agents, are cholinesterase inhibitors. These synthetic compounds are resistant to natural degradation and threaten the environment. We constructed a strain of Pseudomonas putida that can efficiently degrade a model organophosphate, paraoxon, and use it as a carbon, energy, and phosphorus source. This strain was engineered with the pnp operon from Pseudomonas sp. strain ENV2030, which encodes enzymes that transform p-nitrophenol into beta-ketoadipate, and with a synthetic operon encoding an organophosphate hydrolase (encoded by opd) from Flavobacterium sp. strain ATCC 27551, a phosphodiesterase (encoded by pde) from Delftia acidovorans, and an alkaline phosphatase (encoded by phoA) from Pseudomonas aeruginosa HN854 under control of a constitutive promoter. The engineered strain can efficiently mineralize up to 1 mM (275 mg/liter) paraoxon within 48 h, using paraoxon as the sole carbon and phosphorus source and an inoculum optical density at 600 nm of 0.03. Because the organism can utilize paraoxon as a sole carbon, energy, and phosphorus source and because one of the intermediates in the pathway (p-nitrophenol) is toxic at high concentrations, there is no need for selection pressure to maintain the heterologous pathway.Keywords
This publication has 38 references indexed in Scilit:
- Improved Degradation of Organophosphorus Nerve Agents and p-Nitrophenol by Pseudomonas putida JS444 with Surface-Expressed Organophosphorus HydrolaseBiotechnology Progress, 2008
- Laboratory evolution of catabolic enzymes and pathwaysCurrent Opinion in Biotechnology, 2005
- Effect of carbon starvation on p-nitrophenol degradation by a Moraxella strain in buffer and river waterFEMS Microbiology Ecology, 2005
- Growth of Escherichia coli Coexpressing Phosphotriesterase and Glycerophosphodiester Phosphodiesterase, Using Paraoxon as the Sole Phosphorus SourceApplied and Environmental Microbiology, 2004
- Gene Cloning, Purification, and Characterization of a Phosphodiesterase from Delftia acidovoransApplied and Environmental Microbiology, 2003
- Measuring β-Galactosidase Activity in Bacteria: Cell Growth, Permeabilization, and Enzyme Assays in 96-Well ArraysBiochemical and Biophysical Research Communications, 2002
- Mineralization of p-nitrophenol by pentachlorophenol-degrading Sphingomonas spp.FEMS Microbiology Letters, 1997
- Ratios of carbon, nitrogen and phosphorus in Pseudomonas fluorescens as a model for bacterial element ratios and nutrient regenerationAquatic Microbial Ecology, 1996
- Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettesGene, 1995
- Specific-purpose plasmid cloning vectors II. Broad host range, high copy number, RSF 1010-derived vectors, and a host-vector system for gene cloning in PseudomonasGene, 1981