Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Bioluminescent assay of Phospholipase C Using A Luminescent Marine Mutant Bacterium Vibrio harveyi M-17

  • Cho, Ki-Woong (Marine nature products Chemistry Laboratory, Korea Ocean Research and Development Institute) ;
  • Mo, Sang-Jun (Marine nature products Chemistry Laboratory, Korea Ocean Research and Development Institute) ;
  • Lee, Hyi-Seung (Marine nature products Chemistry Laboratory, Korea Ocean Research and Development Institute) ;
  • Park, Jung-Rae (Marine nature products Chemistry Laboratory, Korea Ocean Research and Development Institute) ;
  • Jongheon Shin (Marine nature products Chemistry Laboratory, Korea Ocean Research and Development Institute)
  • Published : 2000.09.01
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Abstract

A bioluminescent assay method for detecting the activity of phospholipase C(PLC; phosphatidyl choline cholinephosphohydrolase, EC 3.1.4.3) was developed using bioluminescent marine bacteria. Phospholipase C from Bacillus cereus and sn-1,2- dimyristoyl glycerol was further hydrolyzed with lipase from Candida ecylidracea. The hydrolyzed myristic acid was quantified using a dark mutant of Vibrio harveyi (designated as M-17). The in vivo light intensity of which was stimulated specifically up to one thousand fold in the presence of myristic acid. The rates of the hdrolysis of the DMPC substrate by the phospholipase measured by the luminescence method were linear with time and the were estalished to detect as little as 0.1 mUnit of phospholipase C and 5 nM of myristic acid production.

Keywords

References

  1. Annu. Rev. Biochem v.56 Inositol triphosphate and diacylglycerol;two interacting secondary messengers Berridge M.J.
  2. Kor. Biochem. J. (presently J. Biochem. Mol. Biol) v.26 A new assay method for bovine serum monoamine oxidase;bacterial luciferase coupled reaction Cho K.W.;S.C. Shim
  3. Kor. J. Microbiol (presently J. Microbiol) v.31 Bioluminescent assay of bovine liver cholesterol esterase using bacterial lucifease coupled reaction Cho K.W.
  4. Kor. Biochem. J. (presently J. Biochem. Mol. Biol) v.27 Bioluminescent assay of phospholipase A₂using luminescent marine bacterium Vibrio harveyi mutant M-17` Cho K.W.
  5. J. Microbiol v.38 Bioluminescent assay of bovine liver ribo-flavin kinase using bacterial luciferase coupled reaction Cho K.W.
  6. J. Biochem. Mol. Biol v.33 Bioluminescent assay of α-oxidase from Cucumins sativas using bacterial luciferase coupled reaction Cho K.W.
  7. Proc. Natl. Acad. Sci. USA v.68 Temperature sensitive mutants in the bacterial bioluminescence system Cline T.W.;J.W. Hastings
  8. The Enzymes Phospholipases Dennis E.A.;P.D. Boyer(ed.)
  9. Enzy. Microb. Technol v.26 Crit-ical evaluation of ρ-nitrophenylphosphorylcholine (ρ-NPPC) as artificial substrate for the detection of phospholipase C. Fliegera A.;S. Gong;M. Faigle;B. Neumeister
  10. Bio-chim. Biophys. Acta v.530 A spectrophoto-metric method for the determination of sphingomyelinase Gatt S.;T. Dinur;Y. Barenholz
  11. Methods Enzymol v.197 Phosphati-dylinositol-specific phosphlipase C from Bacillus cereus and Bacillus thuringiensis Methods Enzymol
  12. Methods Enzymol v.57 Bacterial luciferase;assay, purification and properties Hastings J.W.;T.O. Baldwin;M.Z. Nicoli
  13. J. Opt. Soc. Am v.53 Total quatum flux of iso-topic source Hastings J.W.;G. Weber
  14. Anal. Biochem v.229 Chromogenic assay for phospholipase C from Bacillus cereus Hergenrother P.J.;M.R. Spaller;M.K. Haas;S.F. Martin
  15. Lab-oratory technique in Biochemistry and molecular biology v.3 Techniques of lipidology, Isolation, analysis and identification of lipids Kates M.;T.S. Work(ed.);E. Work(ed.)
  16. Anal. Bio-chem v.75 Phospholipase assay using p-nitrophenylphosphoryl choline and its applications to studying the metal and detergent requirement of the enzyme Kurloka S.;M. Matsuda
  17. Lipase: Structure, mechanism and genetic engineering Lipase and water Legoy M.D.;M. Goldberg;J.N. Barbotin;L. Alberghina(ed.);R.D. Schmid(ed.);R. Verger(ed.)
  18. Anal. Biochem v.39 A stable inexpensive solid state photomultiplier photometer Mitchell G.;J.W. Hastings
  19. Thin-layer chromatography with flame ion-ization detection Ranny M.
  20. Methods Enzymol v.197 Assays of phosphoinositide-specific phospolipase C and purification isozymes from bovine brains Rhee S.G.;S.H. Ryu;K.Y. Lee;K.S. Cho
  21. Biophys. Acta v.1125 Synthetic phos-pholipids as substrates for phospholipase C from Bacillus cereus Ries U.;E.A. Fleer;C. Unger;H. Eibl
  22. Bioorg. Med. Chem. Lett v.9 Syntheis of a new fluorogenic sub-strate for the continous assay of mamalian phosphoinositide-specific phospholipase C Rukavishnikov A.V.;T.O. Zaikova;G.B. Birrell;J.F.W. Keana;O.H. Griffith
  23. Anal. Bio-chem v.214 A chemiluminescent for the detection of phosphatidyl inositol-specific phospholipase C. Ryan M.;J.-H. Huang;O.H. Griffith;J.F. W. Keana;J.J. Volwerk
  24. Anal. Biochem v.198 A fluorescent substrate for the continuous assay of phos-phatidylinositol-specific phospholipase C;synthesis and appli-cation of 2-naphthyl myo-inositol-1-phosphate Shashidhar M.S. J.J. Volwerk;J.F. Keana;O.H. Griffith
  25. Anal. Biochem v.164 A continuous spectrophotometric assay for the Bacillus cereus phospholipase C using a thiophosphate ana-log;evaluation of assay conditions and chrmogenic agent Snyder W.R.
  26. Microbiol. Rev v.57 Bacterial phospholipase C. Titball R.W.
  27. Proc. Natl. Acad. Sci. USA v.75 Myristic acid stimulation of bacterial bioluminescene in aldehyde mutants Ulizer S.;J.W. Hastings