Chiral Separation of Tryptophan by Immobilized BSA(bovine serum albumin) Membrane

BSA고정막에 의한 Tryptophan 이성질체의 분리

  • Kim Min (Department of safety environmental system engineering, Dongguk university) ;
  • Kim Jae-Hun (Department of safety environmental system engineering, Dongguk university) ;
  • Na Won-Jae (Department of safety environmental system engineering, Dongguk university) ;
  • Kim Byoung-Sik (Department of chemical engineering, Dongguk university)
  • 김민 (동국대학교 안전환경시스템공학과) ;
  • 김재훈 (동국대학교 안전환경시스템공학과) ;
  • 나원재 (동국대학교 안전환경시스템공학과) ;
  • 김병식 (동국대학교 생명 화학공학과)
  • Published : 2006.06.01

Abstract

A hollow fiber membrane containing BSA as ligand was Prepared by radiation-induced grafting GMA onto a porous polyethylene hollow fiber and subsequent reacting with DEA and TEA. The density of the DEA and TEA of the membrane were 3.4 mmol/g, 1.7r mmol/g, respectively. The DEA membrane exhibited a higher amount of than the TEA membrane. BSA was immobilized by the graft chains during the permeation of BSA solution throught the DEA and TEA membrane. The BSA was adsorbed in multilayer binding of 8 onto the DEA membrane whereas adsorption onto the TEA membrane remained constant. A two-stage stepwise BTC was observed due to independent chiral recognition for L, D-Trp solution by DEA-BSA membrane.

방사선 그라프트 중합법을 적용하여, 폴리에틸렌 다공성 중공사막에 전자선을 조사시킨 후, glycidyl methacrylate(GMA)를 그라프트 중합하였다. 그 후, 음이온 교환기로서 diethylamine (DEA), triethylamine (TEA)를 도입시켜 2종류의 음이온 교환막을 합성하였다. DEA막과 TEA막의 이온교환 밀도는 3.4 mmol/g, 1.74 mmol/g으로 DEA막이 TEA막보다 높은 이온교환기를 얻을 수 있었다. 이 2종류의 음이온교환막에 단백질(bovine serum albumin, BSA)을 투과법에 의해 고정시켜 BSA 고정막을 만들었다. DEA-BSA막의 경우, 그라프트 체인에 BSA가 8층 이상으로 다층 흡착하였으나, TEA-BSA막의 경우, 강한 음이온에 의해 다층 흡착이 이루어지지 않았다. DEA-BSA막의 경우, BSA 다층 흡착성 고정을 나타내기 때문에 L-Trp가 D-Trp보다 더 강한 흡착 특성을 나타내었다. L, D-Trp 이성질체 혼합물을 투과시킨 BTC에 있어서, DEA-BSA 막의 경우, BSA에 대한 L-Trp와 D-Trp의 키랄 인식이 다르기 때문에 2단계의 BTC곡선을 얻을 수 있었다.

Keywords

References

  1. Nihon Kagaku Kai ed.: Kikan Kagaku Souetus. 6th ed. p2 Gakkai Shuppan Center, Tokyo, Japan (1989)
  2. H. Nakamura, 'Chral Separations by HPLC', Hirokawa Publishing Co., Tokyo, Japan (1997)
  3. J. Haginaka, and N. Kanasugi, 'Enantioselectivity of bovine serum albumin-bonded columns produced with isolated protein fragments', J. Chromatogr., A 694, 71 (1995)
  4. J. Yang and D. S. Haga, 'Role of binding capacity versus binding strength in the separation of chiral compounds on protein-based high-performance liquid chromatography columns Interactions of -and -tryptophan with human serum albumin', J. Chromatogr., A 725, 273 (1996)
  5. H. Hofstetter, O. Hofstetter, and V. Schurig, 'Rapid separation of enantiomers in perfusion chromatography using a protein chiral stationary phase', J. Chromatogr. A 764, 35(1997)
  6. N. Kubota, S. Miura, K. Saito, K. Sugita, K. Watanabe, and T. Sugo, 'Comparison of protein adsorption by anion-exchange interaction onto porous hollow-fiber membrane and gel bead-packed bed', J. Membrane Sci., 117, 135 (1996) https://doi.org/10.1016/0376-7388(96)00054-3
  7. H. Iwata, K. Saito, S. Furusaki, T. Sugo, and J. Okamoto, 'Adsorption characteristics of an immobilized metal affinity membrane', Biotechno. Prog., 7, 412 (1991) https://doi.org/10.1021/bp00011a005
  8. S. Matoba, S. Tsuneda, and K. Saito, and T. Sugo, 'Highly efficient enzyme recovery using a porous membrane with immobilized tentacle polymer chains', Bio/Technology, 13, 795 (1995) https://doi.org/10.1038/nbt0895-795
  9. M. Nakamura, S. Kiyohara, K. Saito, K. Sugita, and T. Sugo, 'High resolution of DL-tryptophan at high flow rates using a bovine serum albumin-multilayered porous hollow-fiber membrane', J. Chromatogr., A 822, 53 (1998)
  10. S. Kiyohara, M. Nakamura, K. Saito, K. Sugita, and T. Sugo, 'Binding of DL-tryptophan to BSA adsorbed in multilayers by polymer chains grafted onto a porous hollow-fiber membrane in a permeation mode', J. Membrane Sci., 152, 143 (1999) https://doi.org/10.1016/S0376-7388(98)00215-4
  11. S. Tsuneda, K. Saito, T. Sugo, and K. Makuuchi, 'Protein adsorption characteristics of porous and tentacle anion-exchange membrane prepared by radiation-induced graft polymerization', Radiat. Phys. Chem., 46, 239-245 (1995) https://doi.org/10.1016/0969-806X(95)00019-T
  12. S. Tsuneda, K. Saito, S. Furusaki, T. Sugo, and I. Ishigaki, 'Water/acetone permeablity of porous hollow-fiber membrane containing diethylamino groups on the grafted polymer branches', J. Membr. Sci., 71, 1-12 (1992) https://doi.org/10.1016/0376-7388(92)85001-Y
  13. M.Nakamura, S. Kiyohara, K. Saito, K. Sugita, and T. Sugo 'Chiral separation of DL-tryptophan using porous membranes containing multilayered bovine serum albumin crosslinked with glutaraldehyde', J. chromatogr. A, 822, 53-58 (1998) https://doi.org/10.1016/S0021-9673(98)00501-9