Production and Purification of Acetylcholinesterase Inhibitor from Pseudomonas sp960903

Pseudomonase sp. 960903에 의한 acetylcholinesterase 억제제의 생산 및 정제

  • 김경자 (순천향대학교 생명과학부)
  • Published : 2000.12.01


To screen agent for the treat-ment of Alzhimers Disease several strains of bacteria producing acetylcholinesterase inhibitor ware isolated from soil. Strain 960903 showed strong acetylcholinesteras inhibitory activity and low butyrylcholinesterse inhibitory activity. The strain 960903 was identified as Pseudomonas sp. Acetylcholinesterase inhibitor ws highly achieved in fermentation medium containing soluble starch 3.0%, glycerol 1.0%, pharmamedia 0.5%, KCI 0.3%, $CaCO_3$ 0.2%, MgS $O_4$..$7H_2$O 0.05%, $KH_2$$PO_4$ 0.05%(pH6.5) at $30^{\circ}C$ for 4 days. Acetylcholinesterase inhibitor was purified by Diaion WA-30($OH^{-}$) column charomatography and cellulose column chromatography. Acetylcholinesterase inhibi-tor showd the maximum wavelength at 205 nm and was soluble in water, acetic acid, ethanol, methanol and dime-thyl sulfoxide. The concentration of 50% inhibition($IC_{50}$) of inhibitor against acetylcholinesterase was 25$\mu\textrm{g}$/ml. The inhibitor was inactivated on heating ar $100^{\circ}C$ fro 15 min and more stable in acidic region than alkaline region.n.


Psendomonas sp;Acetylcholinesterase inhibitor;Alzheimer's disease


  1. Science v.219 Alzheimer's disease; A disorder of cortical cholinergic innervation Coyel, J.T.;D.L. Price;M. R. Long
  2. Helicopverpazea. Entomol. Exp. Appl. v.65 Toxicity and anticholinesterase activity of the fungal metabolites territrems to the corn earworm Dowd, P. F.;F. C. Peng;J. W. Chen,;K. H. Ling
  3. Beygey's Manual of Systematic Bacteriology v.1 Krieg, N. R.;J. C. Holt
  4. Science v.201 Human serial learning : Enhanement with arecoline and impairment with scopolamine Sitaran, N.;H. Weingartner;J. C. Gillin.
  5. American Society for Microbiology Manual of Methods for General Bacteriology Gerhardt, P. G.;G. E. Murray;R. N. Costilow;E. W. Nester;W. A. Wood;N. R. Krieg;G. B. Philips
  6. Nature. v.331 Novel precurosor of Alzhimer's disease amyloid protein shows protease inhibitory activity Kitaguchi N, Y. Takahashi, Y. Tokushima, S. Shiojiri;H. Ito
  7. Arch. Neurol. v.44 Cholinesterase in cerebrosponal fluid. A longitudinal study in Alzheimer disease Elble, R.;E. Giavobini,;G. F. Scarsella
  8. Lancet v.I Necropsy evidence of ecntral cholinergic deficits in senile dementia Perry, E. K., R. H. Perry, G. Blessed;B, E, Tomlinson.
  9. Science. v.235 Characterzation and chomosomal localization of a cDNA ending brain amyliod of Alzheimer's disease. Goldgaber, D.;M. I. Lerman;W. McBride;U. Saffiotti;D. C. Gajdusek.
  10. Arch. Pathol. Lab. Med. v.107 Alzheimer's disease: The commonest form of amyloidosis. Glenner, G.
  11. Biochem. Pharmacol v.41 In vitro effects of various cholinesterase inhibitors on acetyl and butyrylcholinesterase of healthy volunteers Thomsen, T.;B. Zendeh;J. P. Fisher;H. Kewitz
  12. Brain. v.99 Neurotransmitter related enzymes and indices of hypoxia in senile dementia and other abitrophies Bowen, D. M.;C. B. Smith;P. WhiteA. N. Davison
  13. J. Antibiot v.49 no.8 Arisugacins A and B, Novel and Selective Acetylcholinesterase Inhibitors from Penicillium sp. FO-4249 Kuno, F.;K. Otoguro;K. Shiomi;Y. Iwai;S. Omura
  14. J. Biol. Chem. v.266 The cholinesterases. Taylor, P.
  15. Arch. Neurol. v.42 Diagnosis of Alzheimer;s disease Khachaturian, Z. S.
  16. N. Engl. J. Med. v.324 Tacrine as a treatment for Alzheimer's dementia Relaman, A. S.
  17. J. Natural Products v.58 Acetylcholinesterase inhibition by territrem B derivatives Peng, F. C.
  18. Lancet. v.II Neurotansmitters and C.N.S. diease Dementia Rossor, M. N.
  19. Br. Med. J. v.288 Neurochemical charateristics of early and late onset types of Alzheimer's diease. Rossor, M. N.;L. L. Iversen;G. P. Reynolds;C. Q. Mountjoy;M. Roth
  20. Nature. v.331 A new A4 amyloid mRNA contains a domain homologous to serine protease inhibitors Ponte P;P. Gonzalez-DeWhitt;J. Schilling;J. Miller;D. Hsu;B. Greenberg;K. Davis;W. Wallace;I. Lieberburg;F. Fuller;B. Cordell
  21. Enzyme. v.42 A suitable methods to monitor inhibition of cholinesterase activities in tissues as induced by reversible enzyme inhibitors Thomsen, T.;H. Kewitz;O. Pleul.
  22. Dementia v.1 Galanthamine hydrobromide in a long-term treatment of Alzheimer's disease. Thomsen, T.;U. Bickel;J. P. Fischer;H. Kewitz
  23. Anal. Biochem. v.166 Microtiter assay for acetylcholinesterase Doctor, B. P.;L. Toker;E. Roth;I. Silman
  24. Nature. v.325 The precursor of Alzheimer's disease amyloid A4 protein resembles a cell surface receptor. Kangm J.;H. G. Lemaire;A. Unterbeck;M. J. Salbaum;C. L. Masters;K. H. Grezeschik;G. Multhaup;K. Beyreuther;B. Muller-Hill
  25. J. Antibitics. v.46 Cyclophostin acetylcholinesterase inhibitor from Streotomyces lavendulae Kurokawa, T.;K. Suzuki;T. Hayaoka;T. Nakagawa
  26. Neuroliol. Aging v.3 A signal detectability analysis of the effect of physistigmine on memory in patients with Alzhimer's disease. Mohs, R. C.;K. L. Davis
  27. Tetrahedron Lett. v.28 The structure of cyclopenin and cyclopenol, metabolic products from Penicillium yeloptium Westling and Penicillium viridicatum Westling Mohammed, Y. S.;M. Luckner
  28. Biochem. Pharmacol. v.7 A new and rapid colorimetric determination of acetylcholinesterace activity Ellman, G. L.;K. D. Coutrney;V. Anders;R. M. Featherstone
  29. J. Antibiot v.48 Arisugacin, a novel and selective inhibitor of acetylcholinesterase from Penicillium sp. FO-4259 Omura, S.;F. Kuno;K. Otoguro;T. Sunazuka;K. Shiorni;R. Masuma;Y. Iwai
  30. J. Neurol. Sci. v.62 Neuropeptides in Alzheimer type dementia Ferrier I. N;A. J. Cross;J. A. Johnson;G. W. Roberts;T. J. Crow;J. Corsellis, Y. C. Lee;T. E. Adrian;G. P. McGergor;A. J. Baracese-Hamilton;S. R. Bloom
  31. Science v.201 Physostigmine; Improvement of long term memory processes in normal humans Davis, K. L.;R. C. Mohs;J. R. Tinklenberg;G. A. Pfeferbaum;L. E, Hollister;B. S. Kopell
  32. Life. Sci. v.46 Selective inhibition of human acetylcholinesterase by galanthamine in vitro and in vivo Thomsen, T.;H. Kewitz
  33. N. Engl. J. Med. v.315 Oral tetrahydroaminoacridine in long term treatment of senile dementia, Alzheimer type Summers, W. K.;L. V. Majovski;G. M. Marsh;K. Tachiki;A. Kling
  34. Lancet. v.I Choline in Alzheimer's diease. Smith, E.
  35. Brain Res. Rev. v.4 The organisation of some projections of cholinergic neurons of the mammalian forebrain Fibiger, H. C.
  36. Neurosci. Lett. v.152 Effect of territrem-B on cholinergic responeses of snail neuron. Arvanov, V. L.;K. H. Ling;R. C. Chen;M. C. Tsai
  37. Nature. v.314 Location of neurnal tangles in somatostatin neurones in Alzheimer;s diease. Roberts, G. W.;T.J. Crow;J. M. Polak.
  38. Proc. Natl. Aced. Sci. v.88 An in vivo model for the neurodegenerative effects of amyloid and protection by substance P. Kowall, N. W.;M. F. Beal;J. Busciglio;L. K. Duffy;B. A. Yankner
  39. Anal. Biochem. v.196 A silica gel plate-based acetylcholinesterase activity: A mass method to screen for potential inhibitors Kiely, J. S.;W.H. Moss;M. R. Pavia;R. D. Schwarz;G. L. Woodard
  40. Br. Med. J. v.II Correlation of cholonergic abnormalities with senile plaques and mental test scroes in senile dementia Perry, E. K.;B.E. Tomlinson;G. Blessed;K. Bergmann;P.H. Gibson;R. H. Perry
  41. Appl. Environ. Microbiol. v.37 Territrems, trernotogenic mycotoxins of Aspergillus terreus Ling, K. H.;C. K. Yang;F. T. Peng
  42. Biol. Psych. v.17 Modest facilitation of memory in dementia with combiend lecitin and anticholinesterase treatment Kaye, W. H.;N. Sitaram;H. Weingartner;M. H. Ebert;S. Smallbetg;G. Gillin