Pseudomonas sp. CB-33이 생산하는 $\beta$-Xylosidase의 특성

  • 유진환 (충북대학교 자연과학대학 미생물학과) ;
  • 김현구 (영동전문대학 호텔조리과) ;
  • 김치경 (충북대학교 자연과학대학 미생물학과) ;
  • 임재윤 (충북대학교 자연과학대학 미생물학과)
  • Published : 1996.04.01

Abstract

The $\beta$-xylosidase was purified 99- fold from the culture supernatant of Pseudo onas sp. CB-33 by ammonium sulfate precipitation, PEI precipita- tion, DEAE-Sephadex column chromatography, Sephadex G-75 gel filtration chromatography and preparative disc gel electrophoresis. Molecular weight of the enzyme was estimated to be 44,000 by SDS polyacrylamide gel electrophoresis. The enzyme has a pH optimum for activity at 7.0 and is stable over pH 6.5-9.0. The optimal temperature of the enzyme was 45$\circ$C, and its enzymatic activity was completely inactivated at 55$\circ$C for 30 min. Km value of the enzyme for p-nitrophenyl-$\beta$-D-xylopyranoside was calculated to be 4.6 mM. The effect of various reagents on the $\beta$-xylosidase activity was investigated. The enzyme activity was completely inhibited by Hg$^{2+}$, Cu$^{2+}$ and Zn$^{2+}$. The $\beta$-xylosidase was inactivated by tryptophan-specific reagent, N-bromosuccinimide and tyrosine-specific reagent, iodine. The enzyme could degrade xylo-oligosaccharides to xylose and the enzyme was competitively inhibited by xylose. The $\beta$-xylosidase and endoxylanase from Psedomonas sp. CB-33 hydrolized xylan synergically. The purified enzyme also showed $\alpha$-L-arabinofuranosidase activity.

Keywords

References

  1. Enzyme Microb. Technol. v.9 Pachysolen tannophilus : Properties and process considerations for ethanol production from D-xyloase Sliniger,P.J.;M.P.L.Bolen;C.P.Kurtzman
  2. Biotechnol. Lett. v.6 Fermentation of D-xylose to ethanol by a strain of Candida shehatae Du Preez,J.C.;J.P.van der Walt
  3. Biotechnol. Bioeng. v.35 Optimum pH and temperature conditions for xylose fermentation by Pichia stipitis Sliniger,P.J.;R.J.Bothast;M.R.Ladish;M.R.Okos
  4. Appl. Environ. Microbiol. v.55 ³¹P nucler magnetic resonance study of the effect of azide on xylose fermentation by Candida tropicalis Lohmeiere-Vogel,E.;K.Skoog;H.Vogel;B.Hahn-H gerdal
  5. Appl. Microbiol. Biotechnol. v.19 The combined enzymatic hydrolysis and fermentation of hemicellulose to 2,3-butanediol Ernest,K.;C.Yu;L.Deschatelets;J.N.Saddler
  6. Biotechnol. Lett. v.13 Xylitol production from D-xylose by Candida guillermond II: Fermentation behaviour Meyrial,V.;J.P.Delgenes;R.Moletta;J.M.Navarro
  7. J. Ferment. Bioeng. v.67 Conversion of D-xylose into xylitol by immobilized cells of Candida pelliculosa and Methanobacterium sp. HU Nishio,N.;K.Sugawa;N.Hayase;S.Nagai
  8. Kor. J. Appl. Microbiol. Bioeng. v.17 Production of xylanase by Bacillus stearothermophilus Song,H.S.;Y.J.Choi
  9. Adv. Biochem. Eng. v.20 Conversion of hemicellulose carbohydrates Gong,C.S.;L.F.Chen;M.C.Flickinger;G.T.Tsao
  10. J. Appl. Biochem. v.5 β-xylosidase from Aspergillus niger 15. Purification and properties Rodionova,N.A.;I.Tavobilov;A.Bezborodov
  11. Agric. Biol. Chem. v.50 Purification and characterization of extracellular β-xylosidase and β-glucosidase from Aspergillus fumigatus Kitpreechavanch,V.;M.Hayashi;S.Nagai
  12. Meth. Enzymol. v.83 β-D-xylosidase from Penicillium wortmanni Deleyn,F.;M.De Bruyne Ck
  13. Agric. Biol. Chem. v.48 Purification and some properties of β-xylosidase from Trichoderma viride Matsuo,M.;T.Yasui
  14. Agric. Biol. Chem. v.49 Purification and some properties of Chaetomium trilaterale β-xylosidase Uziie,M.;M.Matsuo;T.Yasui
  15. J. Biol. Chem. v.153 A photometric adaptation of the Somogyi method for the determination Nelson,N.
  16. J. Biol. Chem. v.153 Protein measurement with folin phenol reagent Lowry,O.H.;Rosebrough,N.J.;Farr,A.L.;Randall,R.J.
  17. Ann. N.Y. Acad. Sci. v.121 Disc electrophoresis II: Method and application to human serum proteins Davis,B.J.
  18. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T₄ Laemmli,U.K.
  19. Carbohydrate analysis: a practical approach v.2 Monosaccharides Chaplin,M.F.;Chaplin,M.F.(ed.);J.F.Kennedy(ed.)
  20. Biotechnol. Appl. Biochem. v.8 Purification and characterization of xylanolytic enzymes from Trichoderma reesei Lappalainen,A.
  21. Kor. J. Appl. Microbiol. Bioeng. v.11 Studies of hemicellulase system in Aspergillus niger(Purification and characterization of β-xylosidase) Lee,H.J.;Y.D.Choi;M.H.Han
  22. Kor. J. Appl. Microbiol. Bioeng. v.15 Purification and properties of extracellular β-xylosidase from thermophilic alkalophilic Bacillus K-17 Sung,N.K.;I.S.Kang;H.K.Chun;T.Akiba;K.Horikoshi
  23. Biochem. J. v.273 Purification and properties of an aryl β-xylosidase from a cellulolytic extreme thermophile expressed in Escherichia coli Hudson,R.C.;L.R.Schofield;T.Coolbear;R.M.Daniel;H.W.Morgan
  24. Biochem. Biophys. Res. Comm. v.170 Xylan degradation by the thermophile Clostridium stercorarium: cloning and expression of xylanase, and α-arabinofuranosidase genes in Escherichia coli Schwarz,W.H.;H.Adelsberger;S.Jauris;C.Hertel;B.Funk;W.L.Staudenbauer
  25. Appl. Microbiol. Biotechnol. v.28 Characteristics of Trichoderma reesei. β-xylosidase and its use in the hydrolysis of solubilized xylans Poutanen,K.;J.Puls
  26. Appl. Environ. Microbiol. v.57 Sequencing and expression of the Butyrivibrio fibrisolvens xylB gene encoding a novel bifunctional protein with β-d-xylosidase and α-L-arabinofuranosidase activities Utt,E.A.;C.K.Eddy;K.F.Keshav;L.O.Ingram
  27. Kor. J. Appl. Microbiol. Biotechnol. v.20 Molecular cloning and expression of Bacillus stearo-thermophilus β-D-xylosidase gene in E. coli Oh,S.W.;S.S.Park;Y.I.Park;Y.J.Choi