Advertisement

Applied Biochemistry and Biotechnology

, Volume 110, Issue 1, pp 11–21 | Cite as

l-Asparaginase release from Escherichia coli cells with aqueous two-phase micellar systems

Article

Abstract

A method was proposed to release and separate l-asparaginase (EC 3.5.1.1) from Escherichia coli ATCC 11303 cells with aqueous two-phase micellar systems. The systems were composed of K2HPO4 and Triton X-100. The method combines enzyme release with enzyme purification. The influence of Triton X-100 concentration, K2HPO4 concentration, and pH on the release and partition of l-asparaginase was investigated. Experimental results showed that E. coli cells treated with 9.4% (w/v) K2HPO4 and 15% (w/v) Triton X-100 at 25°C for 15–20 h released nearly 80% of the enzyme. Most of the released enzyme was partitioned to the bottom phase (phosphate-rich phase). The effects of Triton X-100 concentration, K2HPO4 concentration, and pH on cloud point were also studied. Electron micrography indicated that the chemical treatment altered the inner structure of E. coli cells significantly.

Index Entries

l-Asparaginase enzyme release Escherichia coli aqueous two-phase micellar system dipotassium hydrogen phosphate Triton X-100 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Lee, S.-M., Ross, J. T., Gustafson, M. E., Wroble, M. H., and Muschik, G. M. (1986), Appl. Biochem. Biotechnol. 12, 229–247.Google Scholar
  2. 2.
    Lee, S.-M., Wroble, M. H., and Ross, J. T. (1989), Appl. Biochem. Biotechnol. 22, 1–11.Google Scholar
  3. 3.
    Hettwer, D. and Wang, H. (1989), Biotechnol. Bioeng. 33, 886–895.CrossRefGoogle Scholar
  4. 4.
    Naglak, T. J. and Wang, H. Y. (1990), Enzyme Microb. Technol. 12, 603–611.CrossRefGoogle Scholar
  5. 5.
    Ariga, O., Miyakawa, I., Aota, T., and Sano, Y. (1994), J. Ferment. Bioeng. 77, 71–74.CrossRefGoogle Scholar
  6. 6.
    Liu, C.-L., Nikas, Y. J., and Blankschtein, D. (1996), Biotechnol. Bioeng. 52, 185–192.CrossRefGoogle Scholar
  7. 7.
    Liu, C.-L., Kamei, D. T., King, J. A., Wang, D. I. C., and Blankschtein, D. (1998), J. Chromatogr. B 711, 127–138.CrossRefGoogle Scholar
  8. 8.
    Sivars, U. and Tjerneld, F. (2000), Biochim. Biophys. Acta 1474, 133–146.Google Scholar
  9. 9.
    Jones, M. N. (1999), Int. J. Pharm. 177, 137–159.CrossRefGoogle Scholar
  10. 10.
    Collén, A., Persson, J., Linder, M., Nakari-Setälä, T., Penttilä, M., Tjerneld, F., and Sivars, U. (2002), Biochim. Biophys. Acta 1569, 139–150.Google Scholar
  11. 11.
    Zhao, F. and Yu, J. (2001), Biotechnol. Prog. 17, 490–494.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  1. 1.School of PharmacyShanghai Jiao Tong UniversityShanghaiP. R. China

Personalised recommendations