Journal of Microbiology and Biotechnology

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

Cloning and Characterization of Cycloinulooligosaccharide Fructanotransferase (CFTase) from Bacillus polymyxa MGL21

  • Jeon, Sung-Jong (The special division for Huma Life Technology, National Institute of Advanced Industrial Science and Technology) ;
  • You, Dong-Ju (Department of Microbiology, Graduate School, Dongeui University) ;
  • Kwon, Hyun-Ju (Department of Material and Life Science, Graduate School of Engineering, Osaka University) ;
  • Shigenori Kanaya (Department of Material and Life Science, Graduate School of Engineering, Osaka University) ;
  • Namio Kunihiro (Department of Material and Life Science, Graduate School of Engineering, Osaka University) ;
  • Kim, Kwang-Hyeon (Department of Microbiology, Graduate School, Dongeui University) ;
  • Kim, Young-Hee (Department of Microbiology, Graduate School, Dongeui University) ;
  • Kim, Byung-Woo (Department of Microbiology, Graduate School, Dongeui University)
  • Published : 2002.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

Microorganism producing extracellular CFTase was isolated from soil and designated as Bacillus polymyxa MGL21. The gene encoding the CFTase (cft) from B. polymyxa MGL21 was cloned and sequenced. The ORF of the cf gene was composed of 3,999 nucleotides, encoding a protein (1,333 amino acids) with a predicted molecular mass of 149,375 Da. Sequence analysis indicated that CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase ($50\%$ identity, 259 amino acids). Furthermore, CFTase possessed a highly conserved core region, which is considered to be functional for the hydrolysis reaction of inulin. The cft gene was expressed in a His-tagged form in Escherichia coli cells, and the His-tagged CFTase was purified to homogeneity. The optimal temperature and pH for CFTase activity were found to be $50^{\circ}C$ and 9.0, respectively. The enzyme activity was completely inhibited by 10 mM $Ag^+\;and\;Cu^2+$. Thin-layer chromatography analyses indicated that CFTase catalyzed not only the cyclization reaction ut also disproportionation and hydrolysis reactions as well.

Keywords

References

  1. J. Bacteriol. v.47 Cloning of the genes for penicillinase, penP and penl, of Bacillus licheniformis in some vector plasmids and their expression in Escherichia coli, Bacillus subtilis and Bacillus licheniformis Imanaka,T.;T.Tanaka;H.Tsunekawa;S.Aiba
  2. J. Microbiol. Biotechnol. v.11 Cloning and sequencing of the β-amylase gene from Paenibacillus sp. and its expression in Saccharomyces cerevisiae Jeong,T.H.;H.O.Kim;J.N.Park;H.J.Lee;D.J.Shin;H.B.Lee;S.B.Chun;S.Bai
  3. Appl. Environ. Microbiol. v.63 Recombinant thermostable cycloinulo-oligosaccharide fructanotransferase produced by Saccharomyces cerevisiae Kanai,T.;N.Ueki;T.Kawaguchi;Y.Teranishi;H.Atomi;C.Tomorbaatar;M.Ueda;A.Tanaka
  4. Biosci. Biotechnol. Biochem. v.57 Reaction catalyzed by cycloinulo-oligosaccharide fructanotransferase Kawamura,M.;T.Uchiyama https://doi.org/10.1271/bbb.57.343
  5. Carbohydr. Res. v.260 Purification and some properties of cycloinulo-oligosaccharide fructanotransferase from Bacillus circulans OKUMZ 31B. Kawamura,M.;T.Uchiyama https://doi.org/10.1016/0008-6215(94)84047-4
  6. Carbohydr. Res. v.192 Formation of a cycloinulo-oligosaccharide from inulin by an extracellular enzyme of Bacillus circulans OKUMZ 31B Kawamura,M.;T.Uchiyama;T.Kuramoto;Y.Tamura;K.Mizutani https://doi.org/10.1016/0008-6215(89)85167-5
  7. J. Ferment. Bioeng. v.79 Gene cloning and characterization of thermostable peptidyl prolyl cis-trans isomerase (PPIase) from Bacillus stearothermophilus Kim,D.J.;M.Morikawa;M.Takagi;T.Imanaka https://doi.org/10.1016/0922-338X(95)94073-Z
  8. J. Microbiol. Biotechnol. v.8 Purification and characterization of cycloinulooligosaccharide fructanotransferase from Bacillus macerans CFC1 Kim,H.Y.;Y.J.Choi
  9. J. Microbiol. Biotechnol. v.10 Regulation of cycloinulooligosaccharide fructanotransferase synthesis in Bacillus macerans and Bacillus subtilis Kim,H.Y.;Y.J.Choi
  10. Appl. Environ. Microbiol. v.67 Molecular characterization of cycloinulooligosaccharide fructanotransferase from Bacillus macerans Kim,H.Y.;Y.J.Choi https://doi.org/10.1128/AEM.67.2.995-1000.2001
  11. J. Microbiol. Biotechnol. v.6 Production of cycloinulooligosaccharide fructanotransferase (CFTase) from Bacillus sp. CFC1. Kim,H.Y.;J.B.Park;Y.M.Kwon;Y.J.Choi
  12. Biosci. Biotechnol. Biochem. v.58 Production of cyclofructan from inulin by Bacillus circulans MCI-2554 Kushibe,S.;R.Sashida;Y.Morimoto https://doi.org/10.1271/bbb.58.1136
  13. J. Microbiol. Biotechnol. v.11 The phylogenetic relationship of several oscillatorian Cyanobacteria, forming blooms at Daecheong reservoirs, based on partial 16S rRNA gene sequences Lee,W.J.;K.S.Bae
  14. J. Biol. Chem. v.153 A photometric adaptation of the Somogyi method for the determination of glucose Nelson,N.
  15. Molecular Cloning: A Laboratory Manual (2nd Ed.) Sambrook,J.;E.F.Fritsch;T.Maniatis
  16. Proc. Natl. Acad. Sci. USA v.74 DNA sequencing with chain-terminating inhibitors Sanger,F.;S.Nicklen;A.R.Couslon https://doi.org/10.1073/pnas.74.12.5463
  17. Bergey's Manual of Systematic Bacteriolology v.2 Smeath,P.H.A.;N.S.Mair;M.E.Sharpe;J.G.Holt
  18. J. Org. Chem. v.59 Binding characteristics of a new host family of cyclic oligosaccharides from inulin: Permethylated cycloinulo-hexose and cycloinulo-heptaose Takai,Y.;Y.Okumura;T.Tanaka;M.Sawada;S.Takahashi;M.Shiro;M.Kawamura;T.Uchiyama https://doi.org/10.1021/jo00090a013
  19. J. Microbiol. Biotechnol. v.10 Purification and characterization of a thermostable β-glycosidase from Thermus caldophilus GK24 Yoo,J.;K.W.Han;H.K.Kim;M.H.Kim;S.T.Kwon
  20. J. Microbiol. Biotechnol. v.11 Co-production of dextran and mannitol by Leuconostoc mesenteroides Yoo,S.K.;D.Kim;D.F.Day
  21. J. Microbiol. Biotechnol. v.11 Effect of His192 mutation on the activity of alginate lyase A1 from Sphingomonas species A1 Yoon,H.J.;Y.J.Choi;O.Miyake;W.Hashimoto;K.Murata;B.Mikami