Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Characterization of C-P Lyase gene cluster by in vivo $^{31}$ P-NMR spectroscopy

  • Lee, Ki-Sung (Department of Biology, Pai-Chai University) ;
  • Kwak, In-Young (Department of Genetic Engineering, Pai-Chai University)
  • Published : 1995.12.01
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Abstract

$\^$31/ P-NMR experiment was performed to detect phophonates (Pn) utilization and degradation in the several different C-P lyase mutants of E. coli and in E. aerogenes and the recombinants. The relative peak intensity (RPI) for the standard samples of 0.5 mM methylphosphonate (MPn) and 1.0 mM aminoethylphosphonate in glucose-MOPS medium showed 0.5 : 1.0 ratio. In the case of BW14329 (.DELTA.phnC-P, .delta.phoA), RPI did not change significantly after 24 hrs culturing, which means it nearly could not utilize Pn. In vivo $\^$31/ P-NMR spectrum of E. aerogens (BWKL 16627) during 3 hrs starvation showed two intense peaks at 0-2 ppm and at near-10 ppm which indicate intracellular orthophosphate (Pi) and pyrophosphate (PPi), respectively. Both of them might be released by degradation of inorganic polyphosphate pool. When MPn is supplied to the medium as an unique P source, Pi content in the cell has the constant, but PPi seems to be slightly decreased. Recombinants (BWKL 16954) grew slower than E. aerogenes in the glucose-MOPS media with various P sources. In vivo $\^$31/ P-NMR spectrum of recombinant did not show any intense signal in the cell. Surprisingly, under the cultivation adding with MPn, a few intense peaks in the region of Pi AND phospate monoester were detected.

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References

  1. Biochim. Biophys. Acta. v.1055 A ³¹P-NMR study of phosphate transport and compartmentation in Candida utilis Bourne,R.M.
  2. J. Bacteriol. v.128 Fosfomycin resistance: Selection method for internal and extended deletion of the phosphoenolpyruvate: Sugar phosphotransferase genes of Salmonella typhimurium Cordaro,J.C.;T.Melton;J.P.Stratis;M.Atagun;C.Gladding;P.E.Hartman;S.Roseman
  3. J. Biol. Chem. v.265 Molecular biology of carbon-phosphorus bond cleavage: Cloning and seqencing of the phn(psiD) genes involved in alkylophosphonate uptake and C-P lyase activity in Escherichia coli B Chen,C.M.;Q.Ye;Z.Zhu;B.L.Wanner;C.T.Walsh
  4. J. Am. Chem. Soc. v.108 Degradation and detoxification of organophosphonates: Cleavage of the carbon to phosphorus bond Cordeiro,M.L.;D.L.Pompliano;J.W.Frost
  5. FEMS Microbiol. Lett. v.5 Biodegradation of phosphonate toxicants yields methane or tehane on cleavege of the C-P bond Daughton,C.G.;A.M.Cook;M.Alexander
  6. Eur. J. Biochem. v.133 Purification and properties of 2-aminoethy;phosphonate: Pyruvate aminotransferase form Pseudomonas aeruginosa Dumora,C.;A.-M.Lacoste;A.Cassaigne
  7. Biochim. Biophys. Acta. v.997 Phosphonacetaldehyde hydrolase from Pseudomonas aeruginosa: Purification properties and comparison with Bacillus cereus enzyme Dumora,C.;A.-M.Lacoste;A.Cassaigne
  8. Biochemistry of natural C-P compounds Horiguchi,M.;T.Hori(eds.);M.Horiguchi(eds.);A.Hayashi(eds.)
  9. J. Bacteriol. v.177 Molecular cloning, mapping, and regulation of Phoregulon gene for phosphonate break down by the phosphonatase pathway of Salmonella typhimurium LT2 Jiang,W.;W.W.Metcalf;K.-S.Lee;B.L.Wanner
  10. Science v.168 Phosphonolipids: Localization in surface membranes of Tetrahymena Kennedy,K.E.;G.A.Thompson
  11. J. Bacteriol. v.174 Evidence for two phosphonate degradative pathways in Enterobacter aerogenes Lee,K.-S.;W.W.Metcalf;B.L.Wanner
  12. J. Bacteriol. v.173 Involvement of the Escherichia coli phn(psiD) gene cluster in assimilation of phosphorus in the form of phosphonate, Pi esters, and Pi Metcalf,W.W.;B.L.Wanner
  13. Gene v.129 Evidence for a fourteen-gene, phnC to P, locus for phosphonate matabolism in Escherichia coli Metcalf,W.W.;B.L.Wanner
  14. J. Bacteriol. v.175 Mutational analysis of an Escherichia coli fourteen-gene operon for phosphonate degradation using TnphoA elements Metcalf,W.W.;B.L.Wanner
  15. J. Bacteriol. v.169 Bacterial carbon-phosphorus lyase: Products, rate, and regulation of phosphonic acid metabolism Wackett,L.P.;S.L.Shames;C.P.Venditti;C.T.Walsh
  16. J. Bacteriol. v.172 Mapping and molecular cloning of the phn(psiD) locus for phosphonate utilization in Escherichia coli Waner,B.L.;J.A.Boline
  17. J. Bacteriol. v.169 The phoBR operon in Escherichia coli K-12 Waner,B.L.;B.-D.Chang