Molecular Cloning and Expression of the Acetyl Xylan Esterase Gene(estII) of Bacillus Stearothermophilus in Escherichia coli

Bacillus stearothermophilus Acetyl Exterase 유전자(estII)의 클로닝과 Escherichia coli에서의 발현

  • Kim, Hee-Sun ;
  • Eom, Soo-Jung ;
  • Cho, Ssang-Goo ;
  • Choi, Yong-Jin
  • 김희선 (고려대학교 유전공학과) ;
  • 엄수정 (고려대학교 유전공학과) ;
  • 조쌍구 (고려대학교 유전공학과) ;
  • 최용진 (고려대학교 유전공학과)
  • Published : 1994.12.01


Bacillus stearothermomophilus, a strong xylan degrader, was confirmed to express multiple esterase activities in addition to the major xylanolytic enzymes. One of the genes encoding the esterases was isolated from the genomic library of B. stearothermophilus constructed with EcoRl restriction endonuclease and pBR322 plasmid. Three recombinant plasmids showing the tributyrin degrading activity were selected from approximately 7, 000 E. coli HB101 transformants, and were found to have the same insert of a 3.2 kb DNA fragment. Restriction mapping and hybridization studies revealed that the gene(estII) on the hybrid plasmid (pKMG7) had originated from the B. stearothermophilus chromosome, and was distinct from the estl, another esterase gene of B. stearothermophilus isolated in the previous work. The E. coli cells harboring pKMG7 produced an acetylxylan esterase that exibited similar substrate specificity to the esterase encoded by the estI gene.


Acetyl xylan esterase;Bacillus stearother-morphilus;molecular cloning


  1. Enzyme Microb. Technol. v.9 α-Glucuronidase in two microbial xylanolytic systems Puls,J.;O.Schmidt;C.Granzow
  2. Microbiol. Rev. v.52 Multiplicity of β-1, 4-xylanase in microorganisms: functions and application Ken,K.Y.Wong;U.L.Larry;J.N.Saddler
  3. Appl. Environ. Microbiol. v.53 Induction of cellulolytic and xylanolytic enzyme systems in Streptomyces spp. MacKenzie,C.R.;D.Bilous;H.Schneider;K.G.Johnson
  4. DNA Cloning v.1 Techniques for transformation of E. coli Hanahan,K.;D.M.Glover(ed.)
  5. Agric. Biol. Chem. v.47 Purification of four esterases from Aspergillus niger Iwai,M.;S.Okumura;E.L. de Leal;Y.Tsujisaka
  6. Recombinant DNA Techniques;An Introduction Rodriguez,R.L;R.C.Tait
  7. Kor. J. Appl. Microbial. Biotech. v.20 Molecular cloning and expression of an endo-xylanase from Bacillus stearothermophilus into Escherichia coli Cho,S.G.;S.S.Park;Y.I.Park;Y.J.Choi
  8. J. Am. Chem. Soc. v.76 The carbohydrates of GramineaseⅡ. The constitution of the hemicelluloses of wheat straw and corn cobs Ehrenthal,I.;R.Montgomery;F.Smith
  9. FEBS Lett. v.186 Acetyl xylan esterases in fungal celluloytic systems Biely,P.;J.Pulse;H.Schneider
  10. Bio/technology v.4 Cooperativity of esterases and xylanases in the enzymatic degradation of acetyl xylan Biely,P.;C.R.MacKenzie;J.Puls;H.Schneider
  11. Kor. J. Appl. Microbial. Biotech. v.21 Molecular cloning and Expression of the acetyl xylan esterase gene of Bacillus stearothermophilus in Escherichia coli Kim,I.S.;S.G.Cho;Y.J.Choi
  12. Biochem. Biophys. Res. Commun. v.141 Molecular cloning and nucleoide sequence of the lipase gene from Pseudomonas fragi Kugimiya,W.;Y.Otani;Y.Hashimoto;Y.Takagi
  13. Molecular Cloning, A Laboratory Manual Maniatis,T.;E.F.Fritsch;J.Sambrook
  14. Meth. Enzymol. v.69 Plasmids of Escherichia coli as cloning vectors Bolivar,F.;K.Backman
  15. Kor. J. Appl. Microbial. Biotech. v.20 Molecular cloning and expression of Bacillus stearothermophilus β-D-xylosidase gene in Escherichia coil Oh,S.W.;S.S.Park;Y.I.Park;Y.J.Choi
  16. J. Chem. Sco. v.12 Constitution of Xylan from Esparto Grass Chanda,S.K.;E.L.Hirst;J.K.N.Jones;E.G.V.Percival
  17. Agric. Environ. v.6 Deacetylation and enhancement of digestibility Bacon,J.S.D.;A.Chesson;A.H.Gordon
  18. Anal. Chem. v.31 Use of dinitrosalicylic acid reagent for determination of reducing sugar Miller,G.L.
  19. Trends Biotechnol. v.3 Microbial xylanolytic systems Biely,P.
  20. Appl. Microbiol. Biotechnol. v.23 Hydrolysis of steamed birchwood hemicellulose by enzymes produced by Trichoderma reesei and Aspergillus awamori Poutanen,K.;J.Puls;M.Linko
  21. Meth. Enzymol. v.160 Measurement of acetyl xylan esterase in Streptomyces Johnson,K.G.;J.D.Fontana;C.R.MacKenzie
  22. Kor. J. Appl. Microbiol. Biotechnol. v.17 Production of xylanase by Bacillus stearothermophilus Song,H.S.;Y.J.Choi
  23. J. Agric. Sci. v.89 The fate of acetyl groups and sugar components during the digestion of grass cell walls in sheep Morris,E.J.;J.S.D.Bacon
  24. Cold Spring Harbor Symp. Quant. Biol. v.43 Complete nucleotide sequence of the Escherichia coli plasmid pBR322 Sutcliffe,J.G.
  25. J. Biol. Chem. v.193 Protein measurement with Folin phenol reagent Lowry,O.H.;N.J.Rasebrough;A.L.Farr;R.J.Randall
  26. Svensk Papperstid v.76 Positions of the O-acetyl groups in pine glucomannan Lindberg,B.;K.G.Rosell;S.Sevensson
  27. Enxyme Microb. Technol. v.14 Enzymic hydrolysis of water-soluble lignin-carbohydrate complexes from Populus deltoides : Effects of combination of β-mannanase, xylanase, and acetyl xylan esterase Ross,N.W.;K.G.Johnson;C.Broun;C.R.MacKenzie;H.Schneider
  28. Carbohydr. Res. v.148 Linkage of ρ-coumaroyl and feruloyl groups to cell wall polysaccharides of barley straw Muller-Harvey,I.;R.D.Hartley;P.J.Harris;E.H.Curzon
  29. Appl. Biotechnol. v.20/21 The role of ester groups in resistance of plant cell wall polysaccharides to enzymatic hydrolysis Gormann,K.;D.J.Mitchell;M.E.Himmel;B.E.Dale;H.A.Schroeder
  30. In the Chemistry of Wood The Hemicelluoses Schuerch,C.
  31. Nucl. Acids Res. v.13 Nonradioactive hybridization probe prepared by the chemical labeling of DNA and RNA with a novel reagent, photobiotin Anthony,C.F.;L.M.James