Journal of Microbiology and Biotechnology

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

Overproduction of Bacillus macerans Cyclodextrin Glucanotransferase in E. coli by Coexpression of GroEL/ES Chaperone

  • Kwon, Mi-Jung (Department of Microbiology, Dong-Eui University) ;
  • So-Lim Park (Department of Microbiology, Dong-Eui University) ;
  • Sung-Koo Kim (Department of Biotechnology & Bioengineering, Pukyong National University) ;
  • Soo-Wan Nam (Department of Biotechnology & Bioengineering, Pukyong National University)
  • Published : 2002.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

The effects of GroEL/ES chaperone on the production of soluble form of B. macerans cyclodextrin glucanotransferase (CGTase) in recombinant E. coli were investigated. The cgt gene and groEL/ES genes are under the control of T7 promoter and Pzt-1 promoter, respectively. The optimal concentrations of inducers, IPTG and tetracycline, were found to be 1.0 mM and 10 ng/ml, respectively. When tetracycline and IPTG were added at the early exponential phase (2h) and exponential phase (3h) of growth, respectively, about 1.5-fold increase of soluble CGTase activity and 1.6-fold increase of soluble CGTase protein were obtained. An SDS-PAGE analysis revealed that about $37.2\%$ of total CGTase protein was in the soluble fraction when GroEL/ES chaperone was overexpressed.

Keywords

References

  1. J. Microbiol. Biotechnol. v.11 Purification and properties of cyclondextring glucanotransferase synthesizing 2-O-α-D-glucopyranosyl L-ascorbic acid from Paenibacillus sp. JB-13 Baw,K.M.;S.K.Kim;I.S.Kong;H.K.Jun
  2. Proc. Natl. Acad. Sci. USA v.89 Cooperation of GroEL/GroES and DnaK/DnaJ heat shock proteins in preventing protein misfolding in Escherichia coli Gragerov,A.;E.Nudler;N.Komissarova;G.A.Gaitanaris;M.E.Gottesman;V.Nikiforov https://doi.org/10.1073/pnas.89.21.10341
  3. Food Sci. Biotechnol. v.8 Enhancement of solubility of Bacillus macerans cyclodextrin glucanotransferase by thioredoxin Han,N.S.;B.Y.Tao
  4. J. Microbiol. Biotechnol. v.11 Effects of environmental factors on in vivo folding of Bacillus macerans cyclodextrin glycosyltransferase in recombinant Escherichia coli Jin,H.H.;N.S.Han;D.K.Kweon;Y.C.Park;J.H.Seo
  5. J. Microbiol. Biotechnol. v.10 Stress responses of the Escherichia coli groE promoter Kwak,Y.H.;S.J.Kim;K.Y.Lee;H.B.Kim
  6. J. Microbiol. Biotechnol. v.10 Constitutive overexpression of the endoxylanase in Bacillus subtilis Kim,J.H.;J.H.Kim;S.C.Kim;S.W.Nam
  7. J. Microbiol. Biotechnol. v.10 Refolding of Bacillus macerans cyclodextrin glucanotransferase expressed as inclusion bodies in recombinant Escherichia coli Kim,C.I.;M.D.Kim;Y.C.Park;N.S.Han;J.H.Seo
  8. J. Biosci. Bioeng. v.90 Improvement of productivity of active horseradish peroxidase in Escherichia coli by coexpression of Dsb proteins Kondo,A.;J.Kohda;Y.Endo;T.Shiromiau;Y.Kurokawa;K.Nishigara;H.Yanagi;T.Yura;H.Fukuda https://doi.org/10.1263/jbb.90.600
  9. J. Biol. Chem. v.267 Effect of overproduction of heat shock chaperones GroESL and DnaK on human procollagenase production in Escherichia coli Lee,S.C.;P.O.Olins
  10. Anal. Biochem. v.181 A spectrophotometric assay for the cyclization activity of cyclomaltohexaose (α-cyclodextrin) glucanotransferase Lejeune,A.K.;Sakaguchi;T.Imanaka https://doi.org/10.1016/0003-2697(89)90385-0
  11. J. Microbiol. Biotechnol. v.11 Isolation of a promoter element that is functional in Bacillus subtills for heterologous gene expression Maeng,C.J.;H.K.Kim;S.Y.Park;B.T.Koo;T.K.Oh;J.K.Lee
  12. Appl. Environ. Microbiol. v.64 Chaperone coexpression plasmics: Differential and assisting folding of an allergen of Japanese cedar pollen, Cryj2, in Escherichia coli Nishihara,K.;M.Kanemori;M.Kitagawa;H.Yanagi;T.Yura
  13. Appl. Environ. Microbiol. v.66 Overexpression of trigger prevents aggregation of recombinant proteins in Escherichia coli Nishihara,K.;M.Kanemori;H.Yanagi;T.Yura https://doi.org/10.1128/AEM.66.3.884-889.2000
  14. J. Microbiol. Biotechnol. v.11 Purification and characterization of Streptococcus mutans cell wall hydrolase from Bacillus subtilis YL-1004 Ohk,S.H.;Y.J.Yoo;D.H.Bai
  15. Food Biotechnol. v.4 Expression of cyclodextrin glucanotransferase from Bacillus macerans in recombinant Escherichia coli Park,Y.C.;C.S.Kim;N.S.Han;J.H.Seo
  16. Starch. v.46 High-level expression of cyclodextrin glucanotransferase in E. coli using a T7 promoter expression system Lee,P.K.C.;B.Y.Tao
  17. Proc. Natl. Acad. Sci. USA v.91 The ATP hydrolysis-dependent reaction cycle of the Escherichia coli Hsp70 system-DnaK, DnaJ, and GrpE. Szabo,A.;T.Langer;H.Schroder;J.Flanagan,B.Bukau;F.U.Hartl https://doi.org/10.1073/pnas.91.22.10345
  18. Cell. v.97 Polypeptide flux through bacterial HsP70: DnaK cooperates with trigger factor in chaperoning nascent chains Teter,S.A.;W.A.Houry;D.Ang;T.Tradler;D.Rockabrand;G.Fischer;P.Blum;C.Georgopoulos;F.U.Hartl https://doi.org/10.1016/S0092-8674(00)80787-4
  19. Appl. Biochem. Biotechnol. v.66 Molecular chaperones, folding catalysts, and the recovery of active recombinant proteins from E. coli. Thomas,J.G.;A.Ayling;F.Baneyx https://doi.org/10.1007/BF02785589
  20. Curr. Opin. Biotechnol. v.6 Effects of overexpressing folding modulators on the in vivo folding of heterologous proteins in Escherichia coli Wall,J.G.;A.Pluckthun https://doi.org/10.1016/0958-1669(95)80084-0
  21. Cell. v.83 Mechanism of GroEL action: productive release of polypeptide from a sequestered position under GroES Weissman,J.S.;C.M.Hohl;O.Kovalenko;Y.Kashi;S.Chen;K.Braig;H.R.Saibil;W.A.Fenton;A.L.Horwich https://doi.org/10.1016/0092-8674(95)90098-5
  22. Cell. v.84 Characterization of the active intermediate of a GroEL-groES-mediated protein folding reaction Weissman,J.S.;H.S.Rye;W.A.Fenton;J.M.Beechem;A.L.Horwich https://doi.org/10.1016/S0092-8674(00)81293-3
  23. J. Biol. Chem. v.268 Both the Escherichia coli chaperone systems, GroEL/GroES and DnaK/DnaJ/GrpE, can reactivate heat-treated RNA polymerase Zimienowicz,A.;D.Skowyra;J.Zeilstra-Ryalls;O.Fayet;C.Georgopoulos;M.Zylicz