미생물학회지 (Korean Journal of Microbiology)

  • 제36권3호
  • /
  • Pages.181-186
  • /
  • 2000
  • /
  • 0440-2413(pISSN)
  • /
  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Clostridium thermocellum JW20가 생성하는 섬유소분해 효소복합체(cellulosome) 구성단백질의 특성에 관한 연구

Characterization of Subunits Dissociated from Cellulosome of Clostridium thermocellum JW20

  • 최상기 (생명공학연구소 미생물공정연구실)
  • 발행 : 2000.09.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Clostridium thermocellum이 생성하는 섬유소분해 효소복합체인 cellulosome은 26개 의 서로 다른 단백질로 구성되어 있으며 그 구성물질로서 calcium을 포함하고 있다. 견고한 구조의 이 복합체에서 구성단백질을 분리하여 그 기능을 연구할 목적으로 이 복합체를 해체 (dissociation)하려 시도하였다. 이 복합체는 calcium을 제거하였을 대 해체되었다. 해체된 구성 단백질들은 MonoQ column chromatogrphy에 의해 구조단백질인 CipA를 포함한 분획, 91 kDa(CelK-tr), 60 kDa 과 57 kDa 단백질로 구성된 분획과 주로 46 kDa(CelA-tr), 또는 71 kDa(CelS-tr) 단백질을 포함하는 분획들로 크게 분리되었다. 대부분의 분획들은 crystalline cellulose 분해 활성을 보였다. 순수 분리된 71 kDa 단백질은 $60^{\circ}C$~$70^{\circ}C$에서 섬 유소 분해시 calcium에 의존적이었으나 46 kDa 단백질은 그렇지 않았다. 46 kDa 단백질은 cellodextrin을 celloviose 및 cellotriose 단위로 절단하며 cellotetraose 로부터 glucose가 생 성되는 것이 관찰되었다. Cellulosome의 섬유소분해 최성 산물이 cellobiose인 것을 고려할 때 개개 구성단백질의 활성이 이 효소 복합체내에서 조절되어 있음을 알 수 있다.

The cellulosome of Clo.~tr~rlil~m tl\ulcornererfnocellum consistmg of 26 dfferent polypeptides contains calcium. The polypeptides dissociated when calcium was removed. Most of dockerill region in the catalytic polypeptides cleavcd during dmociation. The dissociated polypeptides were well separated by MonoQ column chromatography into CipA containing fraction, a fraction still complexed wit11 91 kDa (CelK-a). 60 IiDa and 57 kDa polypeptides, and fractious contailling mainly single polypeptide of 46 kDa (CelA-a) or 71 1d)a polypeptide (CelS-trj Most or the fractions hydrolyzed c~ystalliue cellulose The purified 71 kDa polypeptide was strictly dependent on calcium for crystalline cellulose hydvolyzing activities a1 $60^{\circ}C$~$70^{\circ}C$ but 46 kDa polypeptide was not. 46 M)a polypeptide digested cellodextri~~ as cellobiose or cellotriose unit, and glucose was produced together with cellobiose and cellotriose froln cellotetraosc. It seems that cellulosome produces final product, cellobiose, through coordinated ~qulation of activities of vannus subunits.

키워드

참고문헌

  1. J. Bacteriol. v.156 Adherence of Clostridum thermocellum to cellulose Bayet,E.A.;R.Kenig;R.Lamed
  2. J. Bacteriol. v.162 Sequence of celluase gene of the thermophilic bacterium Clostridum thermocellum Beguin,P.;P.Cornet;J.P.Aubert
  3. Biochemistry v.35 Dissociation of the cellulosome of Clostridium thermocellum in the presence of ethlene diaminetelraacetic acid occurs with the formation of truncated polypeptides Choi,S.K.;L.G.Ljungdahl
  4. Biochemistry v.35 Structural role of calcium for the organization of the cellulosome of Clostridium thermocellum Choi,S.K.;L.G.Ljungdahl
  5. Microbial enzymes and biotechnology Cellulose degradation by fungi Coughlam,M.P.;W.M.Fogarty(ed.);C,T,Kelly(ed.)
  6. The prokaryotes, 2nd ed. Cellulose-decomposing bacteria and their enzyme systems. i Coughlan,M.P.;F.Mayer;A.B.Balows(ed.)
  7. Proc. Natl. Acad. Sci. USA v.91 Cl-Cx revisited: Intramolecular synergism in a celluase Din,N.;H.G.Damude;N.R.Gikes;R.C.Jr.Miller;R.A.Warren;D.J.Kiburn
  8. Ann. Rev. Microbiol. v.47 The cellulosome: the exocellular organelle of Clostridum Felix,C.R.;L.G.Ljungdahl
  9. FEMS Microbiol. Lett. v.94 Cloning of a Clostridium thermocellum DNA fragment encoding polypeptides that bind the catalytic components of the cellulosome Fujino,T.;P.Beguin;J.P.Aubert
  10. Mol. Microbiol. v.8 Sequencing of a Clostridum thermocellum gene(cipA)encoding the cellulosomal SI protein reveals an unusual degree of internal homology Gergross,V.T.;P.M.Romaniec;N.S.Huskisson;A.L.Deman
  11. FEMS Microbiol. Lett. v.51 A catalogue of Clostridum thermocellum endoglucanase. β-glucosidase and xylanase genes cloned in Eschericha coli Hazlewood,G.P.;M.P.N.Romamee;K.Davidson;O.Grepinet;P.Beguin;J.Millet;O.Raynaud;J.P.Aubert
  12. J. Biochem. Biophys. Meth. v.11 Determination of reducing sugars in the nanomole range with tetrazolum blue Jue,C.K.;P.N.Lipke
  13. J. Bacterol v.181 Cloning and sequence analysis of a new cellulase gene encoding CelK, a majo cellulosome component of Clostridium thermocellum evidence for gene duplication and recombination Kataeva,I.;X.L.Li;H.Chen;S.K.Choi;L.G.Ljungdahl
  14. Appl. Environ. Microbiol. v.56 Subunit composition and glycoside activities of the cellulase complex from Clostridium thermocellum JW20 Kohrung,S.;J.Wiegel;F.Mayer
  15. Proc. Natl. Acad. Sci. USA v.92 The anchorage function of CipA(Cell).a scaffolding protein of the Clostridum thermocellum cellulosome Kruus,K.;A.C.Lua;A.L.Demain;J.H.D.Wu
  16. J. Bacteriol. v.177 Exoglucanase activities of the recombinant Clostridium thermocellum CelS, a major cellulosome component Kruus,K.;W.K.Wang;J.Ching;J.H.D.Wu
  17. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmh,U.K.
  18. Lipids v.1 Chromatographic and electrophoertic technigucs Lake,B.D.;H.J.Goodwin;I.Smith(ed.);J.W.T.Seakins(ed.)
  19. Methods Enzymol v.160 Macromolecular complexes and yellow affinity substance from Clostridum thermocellum Ljungdahl,L.G.;M.P.Coughlan;F.Mayer;Y.Mori;K.Honami
  20. Appl. Environ. Microbiol. v.53 Macromolecular organization of the cellulolytic enzyme complex of Clostridium thermocellum as revealed by electron microscopy Mayer,F.;M.P.Coughlan;Y.Mori;L.G.Ljungdahl
  21. J. Bacteriol v.173 Isolation and properties of a major cellobiohydrolase from the celluosome of Clostridum thermocellum Morag,E.;I.Halevy;E.A.Bayer;R.Lamed
  22. Biochimie v.63 Purification and properties of an endo-β-1,4-glucanase from Clostridium thermocellum Petre,J.;R.Longin;J.Millet
  23. Appl. Environ. Microbiol. v.51 Properties of a Clostridium thermocellum endoglucanase produced in Eschericha coli Schwarz,W.H.;F.Grebrutz;W.L.Staudenbaucr
  24. FEBS Lett. v.291 Interaction of the duplicated segment carried by Clostridum thermocellum cellulase with cellulosome components Tokathds,K.;S.Salamitou;S.P.Beguin;P.Dhurjati;J.P.Aubert
  25. J. Bacteriol v.171 Cloning and DNA sequencing of the gene coding for Clostridium thermocellun cellulase Ss(CelS), a major cellulosome component Wang,W.K.;K.Kruus;J.H.D.Wu
  26. Biochem. J. v.121 The cellulase of Fusarium solani Purification and specificity of the β-1,4-D-glucanase and β-glucosidase components Wood,T.M.
  27. Biochemistry v.27 Two components of an extracellular protein aggregate of Clostridium thermocellum together degrade crystalline cellulose Wu,J.H.D.;W.H.D.Johnson;A.L.Demain