Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Preparation of Enzyme Source for Screening of Enzyme Inhibitor of $\beta$-1,3-glucan Synthase

베타-1,3-글루칸 합성효소 저해제의 스크리닝을 위한 효소원 제조법

  • 박희문 (충남대학교 미생물학과) ;
  • 이동원 (충남대학교 미생물학과) ;
  • 송미령 (충남대학교 미생물학과) ;
  • 김정윤 (충남대학교 미생물학과) ;
  • 김성욱 (KIST 유전공학연구소 생물소재그룹) ;
  • 복성해 (KIST 유전공학연구소 생물소재그룹)
  • Published : 1995.06.01
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Abstract

Assay conditions for screening of $\beta$-1,3-glucan synthase inhibitor were evaluated. Cells in the beginning of mid-log phase showed the highest activity of the $\beta$-1,3-glucan synthase. Cells permeabilized with 1% digitonin treatment could be used as a good crude enzyme source for convenient screening of the $\beta$-1,3-glucan synthase inhibitors. Calcofluor white (0.125% in final) and papulacandin B (25 $\mu$g/ml) inhibit 90% and more than 50% of the $\beta$-1,3-glucan synthase activity, respectively. Cells grown at 37$\circ$C showed higher enzyme activity than those of 25$\circ$C. Catalytic factor of the $\beta$-1,3-glucan synthase was solubilized from particulated membrane preparations, holoenzyme, by extracting with 0.00938% CHAPS.

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References

  1. Science v.264 Human mycoses: Drugs and targets for emerging pathogens Georgopapadakou,N.H.;T.J.Walsh
  2. Trends in Biotechnology v.10 Antifungal drug development: the identification of new targets Tuite,M.F.
  3. Pure & Appl. Chem. v.63 Carbohydrates as structural constituents of yeast cell wall and septum Cabib,E.;S.J.Silverman;J.A.Shaw;S.D.Gupta;H.M.Park;J.T.Mullins;P.C.Mol;B.Bower
  4. J. Bacteriol. v.119 Effect of polyoxin D on chitin synthesis and septum formation in Saccharomyces cerevisiae Bowers,B.;G.Levin;E.Cabib
  5. Ann. Rev. Microbiol. v.47 Genetics and molecular biology of chitin synthesis in fungi Bulawa,C.E.
  6. Antimicrob. Agents Chemother v.35 Differential inhibition of chitin synthethases 1 and 2 from Saccharomyces cerevisiae by polyoxin D and nikkomycins Cabib,E.
  7. J. Bacteriol. v.176 Nikkomycin Z is a specific inhibitor of Saccharomyces cerevisiae chitin synthase isozyme Chs3 in vivo and in vitro Gaughran,J.P.;M.H.Lai;D.R.Kirsch;S.J.Silverman
  8. Kor. J. Appl. Microbiol. Biotechnol. v.23 no.3 Properties of β-1.3-glucan synthase system in Saccharomyces cerevisiae Park,H.M.;J.Y.Kim;S.W.Kim;S.H.Bok
  9. J. Bacteriol. v.152 Activation of chitin synthetase in permeabilized cells of a Saccharomyces cerevisiae mutant lacking proteinase B Fernandez,M.P.;J.U.Correa;E.Cabib
  10. J. Bacteriol. v.163 Effect of Calcofluor White and Congo red on fungal cell wall morphogenesis: in vivo activation of chitin polymerization Roncero,C.;A.Duran
  11. Exp. Mycol. v.8 Calcofluor white inhibits Neurospora chitin synthase activity Selitrennikoff,C.P.
  12. Eur. J. Biochem. v.97 Papulacandin B: an inhibitor of glucan synthesis in yeast spheroplasts Baguley,B.C.;G.Rommele;J.Gruner;W.Wehrli
  13. Proc. Natl. Acad. Sci. USA v.83 Regulation of fungal cell wall growth: a guanidine nucleotide-binding, proteinaceous component required for activity of (1-3)-β-D-glucan synthase Kang,M.S.;E.Cabib
  14. Exp. Mycol. v.12 β(1-3) glucan synthase of Neurospora crassa: solubilization and partial characterization Quigly,D.R.;M.Hrmova;C.P.Selitrenikoff