Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Characterization of the ${\beta}-Cyclodextrin$ Glucanotransferase Bacillus firmus var. alkalophilus and Its Expression in E. coli

  • Park, Tae-Hyung (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Shin, Hyun-Dong (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Lee, Yong-Hyun (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University)
  • Published : 1999.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

The ${\beta}-CGTase$ gene of alkalophilic Bacillus firmus var. alkalophilus was cloned into E. coli using $pZErO^{TM}-2$ as a vector. The cloned gene encoded a total of 710 amino acid residues consisting of 674 amino acids of the matured protein and 36 amino acids of the signal peptide, including 20 amino acids from the lacZ gene in the vector. Although the cloned ${\beta}-CGTase$ gene did not contain the promoter and start codons, it was expressed by the lac promoter and lacZ start codon in the $pZErO^{TM}$ vector. A comparison was made with the amino acid sequence and ten other CGTases from Bacillus sp. Also, ten highly conserved regions, which are important amino acid residues in catalysis of CGTase, were identified. The lac promoter used for expression of the ${\beta}-CGTase$ gene was induced constitutively in recombinant E. coli even without IPTG possibly because of a lack of the lacI gene in both host and vector, repressing the lacZ gene in the lac operon. Its expression was catabolically repressed by glucose, however, its repression was reduced by soluble starch, mainly because of the extremely high increase of the cAMP level. ${\beta}-CGTase$ can be overproduced in the recombinant E. coli by maintaining intracellular cAMP levels mostly through the intermittent feeding of glucose during cultivation.

Keywords

References

  1. Gene v.47 Cycrodextrin glycosyltransferase from Klebsiella pneumoniae M5al: Cloning nucleotide sequence and expression Binder, F.;O. Huber;A. Bock
  2. Nucleic Acids Res. v.7 A rapid alkaline extraction procedure for screening recombinant plasmid DNA Birnboim, H. C.;J. Doly
  3. Kor. J. Appl. Microbiol. Biotechnol. v.21 Screening of new alkalophilic strain for overproduction of cyclodextrin glucanotransferase Do, E. J.;J. B. Park;Y. H. Lee
  4. J. Microbiol. Biotechnol. v.3 Selection and characterization of catabolite repression resistant mutant of Bacillus firmus var. alkalophilus producing cyclodextrin glucanotransferase Do, E. J.;H. D. Shin;C. Kim;Y. H. Lee
  5. J. Bacteriol. v.174 Analysis of mutations in cyclodextrin glucanotransferase from Bacillus stearothermophilus which affect cyclization characteristics and thermostability Fujiwara, S.;H. Kakihara;K. Sakaguchi;T. Imanaka
  6. PCR Technology-Current Innovations Griffin,H.G.;A.M.Griffin
  7. PCR Technology-Current Innovations Griffin, H. G.;A. M. Griffin
  8. Antibodies-A Laboratory Manual Halow, E.;D. Lane
  9. Nucleic Acids Res. v.18 Nucleotide sequence of cyclodextrin glycosyltransferase gene, cgtA. from Bacillus licheniformis Hill, D. E.;R. Aldape;J. D. Rozzell
  10. J. Gen. Microbiol. v.134 Molecular cloning and nucleotide sequence of the cyclomaltodextrin glucanotransferase gene from the alkalophilic Bacillus sp. strain no. 38-2 Kaneko,T.;T.Hamamoto;K.Horikoshi
  11. J. Gen. Microbiol. v.135 Construction of chimeric series of Bacillus cyclodextrin glucanotransferase and analysis of the thermal stabilities and pH optima of the enzymes Kaneko, T.;K. B. Song;T. Hamamoto;T. Kudo;K. Horikoshi
  12. J. Gen. Microbiol. v.134 Molecular cloning and nucleotide sequence of the cyclomaltodextrin glucanotransferase gene from the alkalophilic Bacillus sp. strain no. 38-2 Kaneko, T.;T. Hamamoto;K. Horikoshi
  13. J. Jpn. Soc. Starch Sci. v.34 Spectrophotometric determination of cyclization activity of β-cyclodextrin-forming cyclodextrin glucanotransferase Kaneko, T.;T. Kato;N. Nakamura;K. Horikoshi
  14. J. Microbiol. Biotechnol. v.5 Selection of the constitutive mutant of Bacillus firmus var. alkalophilus and its characteristics of cylodextrin glucanotransferase production Kim, C.;H. D. Shin;Y. H. Lee
  15. Appl. Microbiol. Biotechnol. v.26 Molecular cloning of β-cyclodextrin synthetase gene from an alkalophilic Bacillus and its expression in Escherichia coli and Bacillus subtilis Kimura, K.;T. Takano;K. Yamane
  16. J. Ferment. Bioeng. v.74 Cloning and sequencing of the gene encoding cyclodextrin glucanotrans ferase from Bacillus sp. KC201 Kitamoto, N.;T. Kimura;Y. Kito;K. Omiya
  17. Enzymes for Carbohydyrate Engineering Cyclodextrin producing enzyme (CGTase) Kobayashi, S.;K. H. Park(ed.);J. F. Robyt(ed.);Y. D. Choi(ed.)
  18. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Lammli, U. K.
  19. J. Mol. Biol. v.236 Nucleotide sequence and X-ray structure of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 in a maltose-dependent crystal form Lawson, C. L.;R. van Montfort;B. Strokopytov;H. J. Rozeboom;K. H. Kalk;G. E. de Vries;D. Penninga;L. Dijkhuizen;B. W. Dijkstra
  20. Microbiology v.143 The relationship between external glucose concentration and cAMP levels inside Escherichia coli: Implications for models of phosphotransferase-mediated regulation of adenylate cyclase Notley-McRobb, L.;A. Death;T. Ferenci
  21. FEBS Lett. v.296 Functional relationships between cyclodextrin glucanotransferase from an alkalophilic Bacillus and alpha-amylases: Site-directed mutagenesis of the conserved two Asp and one Glu residues Nakamura, A.;K. Haga;S. Ogawa;K. Kuwano;K. Kimura;K. Yamane
  22. Biochemistry v.33 Four aromatic residues in the active center of cyclodextrin glucanotransferase from an alkalophilic Bacillus sp. 1011: Effect of the replacement on substrate binding and cyclization characteristics Nakamura, A.;K. Yamane
  23. Kor. J. Appl. Microbiol. Biotechnol. v.20 Purification and characterization of cyclodextrin glucanotransferase excreted from Bacillus sp. E1. Park, C. S.;E. J. Woo;S. U. Kuk;B. C. Seo;K. H. Park;H. Lim
  24. Annu. Rev. Genet. v.13 Regulatory sequences involved in promotion and termination of RNA transcription Rosenberg, M.;D. Court
  25. Biochim. Biophys. Acta. v.72 Preparation of transforming DNA by phenol treatment Saito, H.;K. Miura
  26. Molecular Cloning: A Laboratory Manual (2nd ed.) Sambrook, J.;E. F. Fritsch;T. Maniatis
  27. Proc. Natl. Acad. Sci. USA v.74 DNA sequencing with chain terminating inhibitor Sanger, F.;S. Nickelen;A. R. Coulson
  28. Kor. J. Appl. Microbiol. Biotechnol. v.26 Purification and characterization of β-cyclodextrin glucanotransferase excreted by Bacillus firmus var. alkalophilus Shin, H. D.;C. Kim;Y. H. Lee
  29. J. Microbiol. Biotechnol. v.9 The roles of tryptophan and histidine residues in the catalytic activities of β-cyclodextrin glucanotransferase from Bacillus firmus var. alkalophilus Shin, H. D.;C. Kim;Y. H. Lee
  30. Appl. Microbiol. Biotechnol. v.35 Cloning and sequencing of a cyclodextrin glucanotransferase gene from Bacillus ohbensis and its expression in Escherichia coli Sin, K. A.;A. Nakamura;K. Kobayashi;H. Masaki;T. Uozumi
  31. J. Bacteriol. v.166 Molecular cloning, DNA nucleotide sequencing, expression in Bacillus subtilis cells of the Bacillus macerans cyclodexrin glucanotransferase gene Takano, T.;M. Fukuda;M. Monma;S. Kobayashi;K. Kainuma;K. Yamane
  32. Enzyme Microb. Tech. v.22 Bacterial cycldextrin glucanotransferase Tonkova, A.
  33. Biotechnol. Lett. v.18 Secretion of heterologous cyclodextrin glycosyltransferase of Bacillus sp. E1 from Escherichia coli Yong, J. S.;J. N. Choi;S. S. Park;C. S. Park;K. H. Park;Y. D. Choi