Isolation of a Pseudomonas monteilli strain with a novel phosphotriesterase.
A Pseudomonas monteilli strain (designated C11) that uses the phosphotriester coroxon as its sole phosphorus source has been isolated. Native PAGE and activity staining identified a single isozyme with significant phosphotriesterase activity in the soluble fraction of the cell. This phosphotriesterase could hydrolyse both coumaphos and coroxon. The hydrolysis product of coroxon, diethylphosphate, and the thion analogue, coumaphos, could not serve as phosphorus sources when added to the growth medium. The majority of the phosphotriesterase and phosphatase activity was contained in the soluble fraction of the cell. Phosphatase activity was inhibited by vanadate as well as by dialysis against the metal chelator, EDTA. Phosphotriesterase activity was not affected by either vanadate or dialysis with EDTA or 1,10-phenanthroline. Phosphotriesterase activity was regulated by the amounts of both phosphate and coroxon in the medium, whereas total phosphatase activity was regulated by phosphate but not coroxon. A lack of hybridisation using a probe against the opd (organophosphate degradation) gene encoding a phosphotriesterase from Flavobacterium sp. ATCC27551 against bulk DNA from P. monteilli C11 suggested that this strain does not contain opd. The work presented here indicates the presence of a novel phosphotriesterase in P. monteilli C11.
Horne
Irene
I
CSIRO Entomology, G.P.O. Box 1700, ACT 2601, Canberra, Australia. irene.horne@ento.csiro.au
Harcourt
Rebecca L
RL
Sutherland
Tara D
TD
Russell
Robyn J
RJ
Oakeshott
John G
JG
D043303:Aryldialkylphosphatase; D001673:Biodegradation, Environmental; D003372:Coumaphos / Q000031:analogs & derivatives* / Q000378:metabolism; D003470:Culture Media; D004950:Esterases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D007306:Insecticides / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011549:Pseudomonas / Q000145:classification / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D012988:Soil Microbiology; D012989:Soil Pollutants / Q000378:metabolism
D003470:Culture Media; D007306:Insecticides; D010710:Phosphates; D012989:Soil Pollutants; D004950:Esterases; D043303:Aryldialkylphosphatase; C007130:coroxon; D003372:Coumaphos