A consortium comprised of two engineered microorganisms was assembled for biodegradation of the organophosphate insecticide parathion. Escherichia coli SD2 harbored two plasmids, one encoding a gene for parathion hydrolase and a second carrying a green fluorescent protein marker. Pseudomonas putida KT2440 pSB337 contained a p-nitrophenol-inducible plasmid-borne operon encoding the genes for p-nitrophenol mineralization. The co-culture effectively hydrolyzed 500 microM parathion (146 mg l(-1)) and prevented the accumulation of p-nitrophenol in suspended culture. Kinetic analyses were conducted to characterize the growth and substrate utilization of the consortium members. Parathion hydrolysis by E. coli SD2 followed Michaelis-Menten kinetics. p-Nitrophenol mineralization by P. putida KT2440 pSB337 exhibited substrate-inhibition kinetics. The growth of both strains was inhibited by increasing concentrations of p-nitrophenol, with E. coli SD2 completely inhibited by 600 microM p-nitrophenol (83 mg l(-1)) and P. putida KT2440 pSB337 inhibited by 1,000 microM p-nitrophenol (139 mg l(-1)). Cultivation of the consortium as a biofilm indicated that the two species could cohabit as a population of attached cells. Analysis by confocal microscopy showed that the biofilm was predominantly comprised of P. putida KT2440 pSB337 and that the distribution of E. coli SD2 within the biofilm was heterogeneous. The use of biofilms for the construction of degradative consortia may prove beneficial.

Pseudomonas putida KT2442 was engineered to use the organophosphate pesticide parathion, a compound similar to other organophosphate pesticides and chemical warfare agents, as a source of carbon and energy. The initial step in the engineered degradation pathway was parathion hydrolysis by organophosphate hydrolase (OPH) to p-nitrophenol (PNP) and diethyl thiophosphate, compounds that cannot be metabolized by P. putida KT2442. The gene encoding the native OPH (opd), with and without the secretory leader sequence, was cloned into broad-host-range plasmids under the control of tac and taclac promoters. Expression of opd from the tac promoter resulted in high OPH activity, whereas expression from the taclac promoter resulted in low activity. A plasmid-harboring operons encoding enzymes for p-nitrophenol transformation to beta-ketoadipate was transformed into P. putida allowing the organism to use 0.5 mM PNP as a carbon and energy source. Transformation of P. putida with the plasmids harboring opd and the PNP operons allowed the organism to utilize 0.8 mM parathion as a source of carbon and energy. Degradation studies showed that parathion formed a separate dense, non-aqueous phase liquid phase but was still bioavailable.

Seven bacterial isolates which degraded the herbicide metamitron (3-methyl-4-amino-6-phenyl-1,2,4-triazin-5-one) were obtained from field-enhanced soil by liquid enrichment culture. All isolates appeared to be identical and a representative, 0246b, was identified as a Rhodococcus sp. by cell wall and fatty acid analyses. This isolate degraded metamitron as the sole source of carbon within 24 h at 25 degrees C and this is the first report of a bacterium capable of growing with metamitron as the sole source of carbon. Metamitron was degraded less rapidly when it was the sole source of both carbon and nitrogen. The rate and extent of degradation was affected by the presence and type of additional sources of carbon and nitrogen in the culture medium. In studies with [14C]-phenyl-labelled metamitron Rhodococcus sp. 0246b partly mineralized the phenyl ring.

No abstract

This sixth book in the series concerned with pesticide chemistry maintains the high standards set by its predecessors both in terms of the content of the individual contributions and the quality of presentation. It concentrates on the single theme of herbicides and comprises nine chapters each written by one or more internationally recognized experts in their field of review. The book begins with an up-to-date classification of the biological properties and uses of the main families of herbicides and plant growth regulators, followed by an excellent discussion of the role of metabolism in determining herbicide selectivity. The following three chapters cover aspects of the chemistry and biochemistry of some specific herbicide groups, and they are followed by a review of an important topic not often covered in books of this type-the assessment of and implications of occupational exposure to herbicides. A chapter on herbicide behaviour in soil concentrates on the topical problems of movement to ground water and enhanced microbial degradation. There is then further discussion of microbial decomposition and a final chapter dealing with their fate and behaviour in surface waters. All in all this is a first class compilation of topical and relevant reviews and makes essential reading for anyone involved in herbicide chemistry. A. Walker