Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Nucleotide Sequence of Cellulolytic Xylanase Gene (bglBC2) from Bacillus circulans

Bacillus circulans 유래 cellulolytic xylanase 유전자(bglBC2)의 염기서열 결정 및 분석

  • Published : 2006.03.01
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Abstract

The nucleotide sequence of the cloned cellulolytic xylanase gene (bglBC2) from B. circulans ATCC21367 was determined. bglBC2 consists of an 1,224 bp open reading frame (ORF) coding for a polypeptide of 407 amino acids with a deduced molecular weight of 45 kDa. The Shine-Dalgarno (SD) sequence (5'-AAAGGAG-3') was found 9 bp upstream of the initiation codon, ATG. A promoter region corresponding closely to the B. subtilis consensus sequence (-35: TTGACA,-10: TATAAT) was detected, the putative -35 and -10 sequences of which were TTTACA and TATACT, respectively. The deduced amino acid sequence of the cellulolytic xylanase showed 97% homology with that of the alkaline $endo-\beta-1,4-glucanase$ from B. circulans KSM-N257, 75% homology with that of the $endo-\beta-1,3-1,4-glucanase$ from B. circulans WL-12, and 45% homology with that of the $endo-\beta-1,4-glucanase$ (cellulase) from Bacillus sp. KSM-330. The bglBC2 sequence was deposited in Gen-Bank under the accession number AY269256.

클로닝된 Bacillus circulans ATCC21367 유래 cellulolytic xylanase 유전자(bglBC2)의 염기서열을 결정 분석하였다. 본 유전자는 1,224 bp의 407개 아미노산을 암호하는 open reading frame (ORF)으로 구성되어 있었으며 염기서열로부터 산출된 유전자의 분자량은 45 kDa으로 효소의 SDS-PAGE로부터 측정된 분자량과 일치하였다. ATG 개시 코돈의 9bp 위쪽에 Shine-Dalgarno (SD) 서열로 추정되는 5'-AAAGGAG-3' 서열이 확인되었고 그 상단에 promoter로 추정되는 -35 서열(TTTACA)과 -10 서열(TATACT)이 위치하고 있었으며, 이는 B. subtilis promoter consensus sequence와 유사하였다. 한편, 이 효소의 아미노산 서열은 이미 보고된 B. circulans KSM-N257의 alkaline $endo-\beta-1,4-glucanase$와는 97%, B. circulans WL-12의 $endo-\beta-1,3-1,4-glucanase$와는 75%, Bacillus sp. KSM-330의 $endo-\beta-1,4-glucanase$ (cellulase)와는 45%의 유사성을 나타내었다. 또한 bglBCS 염기서 열의 정보를 GenBank에 등록하였으며 등록번호는 Ar269256이다.

Keywords

References

  1. Ashita, D., J.K. Gupta, and S. Khanna. 2000. Enhanced production, purification and characterization of a novel cellulase-poor thermostable, alkalitolerant xylanase from Bacillus circulans AB16. Process Biochem. 35, 849-856 https://doi.org/10.1016/S0032-9592(99)00152-1
  2. Bernier, R.Jr., H. Driguez, and M. Desrochers. 1983. Molecular cloning of a Bacillus subtilis xylanase gene in Escherichia coli. Gene 26, 59-65 https://doi.org/10.1016/0378-1119(83)90036-7
  3. Bhat, M.K. and S. Bhat. 1997. Cellulose degrading enzymes and their potential industrial applications. Biotechnol. Adv. 15, 583-620 https://doi.org/10.1016/S0734-9750(97)00006-2
  4. Brice, R.E. and I.M. Morrison. 1982. The degradation of isolated hemicellulose and lignin-hemi-cellulose complex by cell-free, rumen hemicellulose. Carbohydr. Res. 101, 93-100 https://doi.org/10.1016/S0008-6215(00)80797-1
  5. Bueno, A., C.R. Vazguez de Aldana, J. Correa, T.G. Villa, and F. del Rey. 1990. Synthesis and secretion of a Bacillus circulans WL-12 1,3-1,4- $beta$ -D-glucanase in Escherichia coli. J. Bacteriol. 172, 2160-2167 https://doi.org/10.1128/jb.172.4.2160-2167.1990
  6. Courtin, C.M., A. Roelants, and J.A. Delcour. 1999. Fractionationreconstitution experiments provide insight into the role of endoxylanases in bread-making. J. Agric. Food Chem. 47, 1870-1877 https://doi.org/10.1021/jf981178w
  7. Gat, O., A. Lapidot, T.I. Alchana, C. Regueros, and Y. Shoham. 1994. Cloning and DNA sequence of the gene coding for Bacillus stearothermophilus T-6 xylanase. Appl. Environ. Microbiol. 60, 1889-1896
  8. Hakamada, Y., K. Endo, S. Takizawa, T. Kobayashi, T. Shirai, T. Yamane, and S. Ito. 2002. Enzymatic properties, crystallization, and deduced amino acid sequence of an alkaline endoglucanase from Bacillus circulans. Biochim. Biophys. Acta. 1570, 174-180 https://doi.org/10.1016/S0304-4165(02)00194-0
  9. Hazlewood, G.P., M.P.M. Romaniec, K. Davidson, O. Grepinet, P. Beguin, J. Millet, O. Raynaud, and J.P. Aubert. 1988. A catalogue of Clostridium thermocellum endoglucanase, $\beta$-glucosidase and xylanase genes cloned in Escherichia coli. FEMS Microbiol. Lett. 51, 231-236 https://doi.org/10.1111/j.1574-6968.1988.tb03002.x
  10. Kim, J.Y., H.B. Kim, and D.S. Lee. 2000. Molecular cloning and expression of a cellulolytic xylanase gene from Bacillus circulans in Escherichia coli. Kor. J. Microbiol. 36, 196-202
  11. Kulkarni, N., A. Shendye, and M. Rao. 1999. Molecular and biotechnological aspects of xylanases. FEMS Microbiol. Rev. 23, 411- 456 https://doi.org/10.1111/j.1574-6976.1999.tb00407.x
  12. McCarthy, A.A., D.D. Morris, P.L. Bergquist, and E.N. Baker. 2000. Structure of XynB, a highly thermostable beta-1,4-xylanase from Dictyoglomus thermophilum Rt46B.1, at 1.8 $\AA$ resolution. Acta Crystallogr. D. Biol. Crystallogr. 11, 1367-1375
  13. McCracken, K.J., M.R. Bedford, and R.A. Stewart. 2001. Effect of variety, the 1B/1R translocation and xylanase supplementation on nutritive value of wheat for broilers. Br. Poult. Sci. 42, 638-642 https://doi.org/10.1080/00071660120088452
  14. McIntosh, L.P., G. Hand, P.E. Johnson, M.D. Joshi, M. Korner, L.A. Plesniak, L. Ziser, W.W. Wakarchuk, and S.G. Withers. 1996. The pKa of the general acid/base carboxyl group of a glycosidase cycles during catalysis: A 13C-NMR study of Bacillus circulans xylanase. Biochemistry 35, 9958-9966 https://doi.org/10.1021/bi9613234
  15. Moran Jr, C.P., N. Lang, S.F.J. LeGrice, G. Lee, M. Stephens, A.L. Sonenshein, J. Pero, and R. Losick. 1982. Nucleotide sequence that signal the initiation of transcription and translation in Bacillus subtilis. Mol. Gen. Genet. 186, 339-346 https://doi.org/10.1007/BF00729452
  16. Ozaki, K., N. Sumitomo, and S. Ito. 1991. Molecular cloning and nucleotide sequence of the gene encoding an endo-1,4-$\beta$-glucanase from Bacillus sp. KSM-330. J. Gen. Microbiol. 137, 2299-2305 https://doi.org/10.1099/00221287-137-10-2299
  17. Siedenberg, D., S.R. Gerlach, K. Schugerl, M.L.F. Giuseppin, and J. Hunik. 1998. Production of xylanase by Aspergillus awamori on synthetic medium in shake flask cultures. Process Biochem. 33, 429-433 https://doi.org/10.1016/S0032-9592(97)00090-3
  18. Sunna, A., and G. Antranikian. 1997. Xylanolytic enzymes from fungi and bacteria. Crit. Rev. Biotechnol. 17, 39-67 https://doi.org/10.3109/07388559709146606
  19. Tang, J.C.T., P. McGray, P. Chen, B. Ferguson, G. Carson, and W. S. Kelley. 1983. Properties of xylanase from Clostridium thermocellum using a trinitrophenyl-xylan assay, p. 287-301. In E.J. Soltes (ed.), Wood and agricultural residues. Academic Press, Inc., Orlando, Fla
  20. Tenkanen, H., J. Plus, and K. Poutanen. 1992. Two major xylanases of Trichoderma reesei. Enzyme Microb. Technol. 14, 566-574 https://doi.org/10.1016/0141-0229(92)90128-B
  21. Velikodvorskaya, T.V., I.Y. Volkov, V.T. Vasilevko, V.V. Zverlov, and E.S. Piruzian. 1997. Purification and some properties of Thermotoga neapolitana thermostable xylanase B expressed in E. coli cells. Biochemistry 62, 66-70
  22. Wakarchuk, W.W., R.L. Campbell, W.L. Sung, J. Davoodi, and M. Yaguchi. 1994. Mutational and crystallographic analyses of the active site residues of the Bacillus circulans xylanase. Protein Sci. 3, 467-475 https://doi.org/10.1002/pro.5560030312
  23. Wong, K.K.Y., and J.N. Saddler. 1992. Application of hemicelluloses in the food, feed, and pulp and paper industries, p. 127-143. In: Coughlen, P.P. and G.P. Hazlewood (eds.), Hemicellulose and hemicellulases. Portland press, London
  24. Wong, K.K.Y., L.U.L. Tan, and J.N. Saddler. 1988. Multiplicity of $\beta$-1,4-xylanase in microorganisms; function and applications. Microbiol. Rev. 52, 305-317
  25. Xie, H., H.J. Gilbert, S.J. Charnock, G.J. Davies, M.P. Williamson, P.J. Simpson, S. Raghothama, C.M. Fontes, F.M. Dias, L.M. Ferreira, and D.N. Bolam. 2001. Clostridium thermocellum Xyn10B carbohydrate-binding module 22-2: the role of conserved amino acids in ligand binding. Biochemistry 40, 9167-9176 https://doi.org/10.1021/bi0106742
  26. Yang, R.C.A., C.R. MacKenzie, and S.A. Narang. 1988. Nucleotide sequence of a Bacillus circulans xylanase gene. Nucleic Acids Res. 16, 7187 https://doi.org/10.1093/nar/16.14.7187