, Volume 39, Issue 1–2, pp 15–22 | Cite as

Ligand-exchange chromatography of amino acid enantiomers and its application to the biotransformation of DL-aspartic acid byPseudomonas dacunhae

  • C. Cheng
  • F. -Y. Lin


The separation of the D and L enantiomers of eighteen essential α amino acids has been investigated by ligand-exchange chromatography (LEC). The effect of column temperature on the retention times and resolution of individual amino acid enantiomers has been studied by varying the temperature from 25 to 50 °C for a mobile phase containing Cu2+ ions. By use of a temperature of 50 °C and Zn2+ in the mobile phase, eight of the eighteen amino acid enantiomers can be resolved sufficiently well for practical application. Only phenylalamine, tyrosine, and tryptophan can be separated by use of Ni2+ as complexation metal at 50 °C. LEC has been used to monitor the decarboxylation of racemic DL-aspartic acid byPseudomonas dacunhae. Analysis of DL amino acid enantiomers in different media was performed at column temperatures of 30 and 50°C by addition of 0.125 mM Cu2+ to the aqueous mobile phase. It was found that the analytical performance is most dependent on the identity of the metal used for complexation; the concentration of the metal was of secondary importance and the column temperature less important still.

Key Words

Column liquid chromatography Ligand exchange Amino acid enantiomers DL-aspartic acid Pseudomonas dacunhae 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    G. Zubay, „Biochemistry”, 2nd Edition, MacMillan, New York, 1988, Chapter 1.Google Scholar
  2. [2]
    M. Bodanszky, D. Perlman, Science163, 352 (1969).Google Scholar
  3. [3]
    W. A. Nugent, T. V. Rajan Babu, M. J. Burk, Science259, 479 (1993).Google Scholar
  4. [4]
    M. Ajioka, S. Yano, K. Matsuda, S. Yoshikawa, J. Am. Chem. Soc.103, 2549 (1981).CrossRefGoogle Scholar
  5. [5]
    Y. N. Belokon', A. G. Bulychev, S. V. Vitt, Y. J. Struchkov, A. S. Batsanov, T. V. Timofeeva, V. A. Tsyryapkin, M. G. Ryzhov, L. A. Lysova, V. I. Bakhmutov, V. M. Belikov, J. Am. Chem. Soc.107, 4252 (1985).Google Scholar
  6. [6]
    N. Esaki, H. Shimoi, H. Tanaka, K. Soda, Biotechnol. Bioeng.34, 1231 (1989).CrossRefGoogle Scholar
  7. [7]
    R. D. Kiss, G. Stephanopoulos, Biotechnol. Bioeng.39, 565 (1992).CrossRefGoogle Scholar
  8. [8]
    A. Dyremark, M. Ericsson, Chromatographia29, 51 (1990).CrossRefGoogle Scholar
  9. [9]
    T. Fukuhara, S. Yuasa, J. Chromatogr. Sci.28, 114 (1990).Google Scholar
  10. [10]
    H. Katoh, T. Ishida, Y. Baba, H. Kiniwa, J. Chromatogr.473, 241 (1989).CrossRefGoogle Scholar
  11. [11]
    G. Jeanneret-Gris, J. Porret, K. Bernauer, Chromatographia20, 449 (1990).Google Scholar
  12. [12]
    V. A. Davankov, S. V. Rogozhin, J. Chromatogr.60, 280 (1971).CrossRefGoogle Scholar
  13. [13]
    V. A. Davankov, S. V. Rogozhin, A. V. Semechkin, T. P. Sachkova, J. Chromatogr.82, 359 (1973).CrossRefGoogle Scholar
  14. [14]
    R. Kuhn, S. Hoffstetter-Kuhn, Chromatographia34, 505 (1992).CrossRefGoogle Scholar
  15. [15]
    R. Kuhn, F. Stoecklin, F. Erni, Chromatographia33, 32 (1992).CrossRefGoogle Scholar
  16. [16]
    P. Gozel, E. Gassmann, H. Michelsen, R. N. Zare, Anal. Chem.59, 44 (1987).CrossRefGoogle Scholar
  17. [17]
    T. Shibatani, T. Kakimoto, J. Kato, N. Nishimura, I. Chibata, J. Ferment. Technol.52, 886 (1974).Google Scholar
  18. [18]
    C. Cheng, L.-H. Huang, J. Chromatogr.555, 272 (1991).CrossRefGoogle Scholar
  19. [19]
    A. F. Cotton, G. Wilkinson, P. L. Gaus, “Basic Inorganic Chemistry”, 2nd Edition, John Wiley & Sons, New York, 1987, Chapter 31.Google Scholar
  20. [20]
    K. Yamamoto, T. Tosa, I. Chibata, Biotechnol. Bioeng.22, 2045 (1980).CrossRefGoogle Scholar
  21. [21]
    M. Salo, H. Vuorela, J. Halmekoski, J. Chromatogr.592, 127 (1992).CrossRefGoogle Scholar
  22. [22]
    R. K. Murray, D. K. Granner, P. A. Mayes, V. W. Rodwell, “Haper's Biochemistry”, 21 st Edition, Appleton & Lange, Connecticut, 1988, Chapter 10.Google Scholar
  23. [23]
    C. Cheng, Proc. Natl. Sci. Counc. ROC(A)15, 446 (1991).Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1994

Authors and Affiliations

  • C. Cheng
    • 1
  • F. -Y. Lin
    • 1
  1. 1.Department of ChemistryChung Yuan Christian UniversityChungliTaiwan, ROC

Personalised recommendations