Biotechnology and Bioprocess Engineering:BBE

  • 제10권6호
  • /
  • Pages.500-504
  • /
  • 2005
  • /
  • 1226-8372(pISSN)
  • /
  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Refolding of Fusion Ferritin by Gel Filtration Chromatography(GFC)

  • Kim, Hyung-Won (Department of Chemical Engineering, Chungnam National University) ;
  • Kim, In-Ho (Department of Chemical Engineering, Chungnam National University)
  • 발행 : 2005.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Fusion ferritin (heavy chain ferritin, $F_H+$ light chain ferritin, $F_L$), an iron-binding protein, was primarily purified from recombinant Escherichia coli by two-step sonications with urea [1]. Unfolded ferritin was refolded by gel filtration chromatography (GFC) with refolding enhancer, where 50 mM Na-phosphate (pH 7.4) buffer containing additives such as Tween 20, PEG, and L-arginine was used. Ferritin is a multimeric protein that contains approximately 20 monomeric units for full activity. Fusion ferritin was expressed in the form of inclusion bodies (IBs). The IBs were initially solubilized in 4 M urea denaturant. The refolding process was then performed by decreasing the urea concentration on the GFC column to form protein multimers. The combination of the buffer-exchange effect of GFC and the refolding enhancers in refolding buffer resulted in an efficient route for producing properly folded fusion ferritin.

키워드

참고문헌

  1. Huh, Y. S. and I. H. Kim (2003) Purification fusion ferritin from recombinant E. coli using two-step sonications. Biotechnol. Lett. 25: 993-996 https://doi.org/10.1023/A:1024005408387
  2. Shin, H. C. (2001) Protein folding, misfolding, and refolding of therapeutic proteins. Biotechnol. Bioprocess Eng. 6: 237-243 https://doi.org/10.1007/BF02931984
  3. Lilie, H., E. Schwarz, and R. Rudolph (1998) Advances in refolding of proteins produced in E. coli. Curr. Opin. Biotechnol. 9: 497-501 https://doi.org/10.1016/S0958-1669(98)80035-9
  4. Fahey, E. M., J. B. Chaudhuri, and P. Binding (2000) Refolding and purification of a urokinase plasminogen activator fragment by chromatography. J. Chromatogr. B 737: 225-235 https://doi.org/10.1016/S0378-4347(99)00360-6
  5. Guise A. D. and J. B. Chaudhuri (2001) Initial protein concentration and residual denaturant concentration strongly affect the batch refolding of hen egg white lysozyme. Biotechnol. Bioprocess Eng. 6: 410-418 https://doi.org/10.1007/BF02932322
  6. Geng, X. D. and X. Q. Chang (1992) High-performance hydrophobic interaction chromatography as a tool for protein refolding. J. Chromatogr. A 599: 185-194 https://doi.org/10.1016/0021-9673(92)85472-6
  7. Guan Y.-X., H.-X. Pan, Y.-G. Gao, S.-J. Yao, and M. G. Cho (2005) Refolding and purification of recombinant human interferon-$\gamma$ expressed as inclusion bodies in E. coli using size-exclusion chromatography. Biotechnol. Bioprocess Eng. 10: 122-127 https://doi.org/10.1007/BF02932581
  8. Lee, G., Y. H. Y. Song, H. T. Chung, M. J. Youn, and S. I. Jang (1991) Differential effects of ferritin on the humoral and cellular immune responses in the mouse. Molecular Cells. 1: 169-175
  9. Kim, K. S., S. R. Chang, and Y. T. Kim (1995) Purification and characterization of recombinant tadpole H-chain ferritin in E. coli. J. Biochem. Mol. Biol. 28: 238-242
  10. Lee, J., H. Y. Seo, E. S. Jeon, O. S. Park, K. M. Lee, C. U. Park, and K. S. Kim (2001) Cooperative activity of subunits of human ferritin heteropolymers in. E. coli. J. Biochem. Mol. Biol. 34: 365-370
  11. Kim, K. S. (1998) Influence of site directed mutagenesis on protein assembly and solubility of tadpole H-chain ferritin. Biotechnol. ioprocess Eng. 3: 67-70 https://doi.org/10.1007/BF02932504
  12. Bradford, M. M. (1976) A rapid and sensitive method for the quantization of protein utilizing the principles of protein- dye binding. Anal. Biochem. 72: 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  13. Laemmli, U. K. (1970) Detection of structural proteins during the assembly of the head of Bacteriophage T4. Nature 227: 680-685 https://doi.org/10.1038/227680a0
  14. Kim, S. W., Y. H. Kim, and J. W. Lee (2001) Thermal stability of human ferritin: concentration dependence and enhanced stability of an N-terminal fusion mutant. Biochem. Biophys. Res. Commun. 289: 125-129 https://doi.org/10.1006/bbrc.2001.5931
  15. Gao, Y. G., Y. X. Guan, S. J. Yao, and M. G. Cho (2002) Refolding of lysozyme at high concentration in batch and fed-batch operation. Korean J. Chem. Eng. 19: 871-875 https://doi.org/10.1007/BF02706982
  16. Lee, J.-H., N.-G, Loc, T.-H. Kwon, and M.-S. Yang (2004) Partitioning of recombinant human granulocyte macrophage colony stimulating factor from plant suspension culture in PEG/sodium phosphate aqueous two-phase systems, Biotechnol. Bioprocess Eng. 9: 12-16 https://doi.org/10.1007/BF02949316
  17. Hong, S. P., C. H. Lee, S. K. Kim, H. S. Yun, J. H. Lee, and K. H. Row (2004) Mobile phase compositions for ceramide III by normal phase high performance liquid chromatography. Biotechnol. Bioprocess Eng. 9: 47-51 https://doi.org/10.1007/BF02949321