Applied Biochemistry and Biotechnology

, Volume 171, Issue 4, pp 954–962 | Cite as

Expression of Low Endotoxin 3-O-Sulfotransferase in Bacillus subtilis and Bacillus megaterium

  • Wenya Wang
  • Jacob A. Englaender
  • Peng Xu
  • Krunal K. Mehta
  • Jiraporn Suwan
  • Jonathan S. Dordick
  • Fuming Zhang
  • Qipeng Yuan
  • Robert J. Linhardt
  • Mattheos Koffas
Article

Abstract

A key enzyme for the biosynthesis and bioengineering of heparin, 3-O-sulfotransferase-1 (3-OST-1), was expressed and purified in Gram-positive Bacillus subtilis and Bacillus megaterium. Western blotting, protein sequence analysis, and enzyme activity measurement confirmed the expression. The enzymatic activity of 3-OST-1 expressed in Bacillus species were found to be similar to those found when expressed in Escherichia coli. The endotoxin level in 3-OST-1 from B. subtilis and B. megaterium were 104–105-fold lower than that of the E. coli-expressed 3-OST-1, which makes the Bacillus expression system of particular interest for the generation of pharmaceutical grade raw heparin from nonanimal sources.

Keywords

3-O-Sulfotransferase Bioengineered heparin Endotoxin free Bacillus subtilis Bacillus megaterium Nonanimal sources 

References

  1. 1.
    Bhaskar, U., Sterner, E., Hickey, A. M., Onishi, A., Zhang, F., Dordick, J. S., et al. (2012). Applied Microbiology and Biotechnology, 93, 1–16.CrossRefGoogle Scholar
  2. 2.
    Liu, H. Y., Zhang, Z. Q., & Linhardt, R. J. (2009). Natural Product Reports, 26, 313–321.CrossRefGoogle Scholar
  3. 3.
    Zhang, Z., McCallum, S. A., Xie, J., Nieto, L., Corzana, F., Jimenez-Barbero, J., et al. (2008). Journal of the American Chemical Society, 130, 12998–13007.CrossRefGoogle Scholar
  4. 4.
    Xu, Y. M., Masuko, S., Takieddin, M., Xu, H. M., Liu, R. P., Jing, J., et al. (2011). Science, 334, 498–501.CrossRefGoogle Scholar
  5. 5.
    Anagnostopoulos, C., & Spizizen, J. (1961). Journal of Bacteriology, 81, 741–746.Google Scholar
  6. 6.
    Phan, T. T., Nguyen, H. D., & Schumann, W. (2006). Protein Expression and Purification, 46, 189–195.CrossRefGoogle Scholar
  7. 7.
    Vary, P. S., Biedendieck, R., Fuerch, T., Meinhardt, F., Rohde, M., Deckwer, W. D., et al. (2007). Applied Microbiology and Biotechnology, 76, 957–967.CrossRefGoogle Scholar
  8. 8.
    Edavettal, S. C., Lee, K. A., Negishi, M., Linhardt, R. J., Liu, J., & Pedersen, L. C. (2004). Journal of Biological Chemistry, 279, 25789–25797.CrossRefGoogle Scholar
  9. 9.
    Puyet, A., Sandoval, H., Lopez, P., Aguilar, A., Martin, J. F., & Espinosa, M. (1987). FEMS Microbiology Letters, 40, 1–5.CrossRefGoogle Scholar
  10. 10.
    Paul, P., Liu, J., Dordick, J. S., & Linhardt, R. J. (2012). Analytical and Bioanalytical Chemistry, 403, 1491–1500.CrossRefGoogle Scholar
  11. 11.
    Golovanov, A. P., Hautbergue, G. M., Wilson, S. A., & Lian, L. Y. (2004). Journal of the American Chemical Society, 126, 8933–8939.CrossRefGoogle Scholar
  12. 12.
    Yang, Y. S., Marshall, A. D., McPhie, P., Guo, W. X., Xie, X., Chen, X., et al. (1996). Protein Expression and Purification, 8, 423–429.CrossRefGoogle Scholar
  13. 13.
    Wu, X. C., Lee, W., Tran, L., & Wong, S. L. (1991). Journal of Bacteriology, 173, 4952–4958.Google Scholar
  14. 14.
    Fu, L. L., Xu, Z. R., Li, W. F., Shuai, J. B., Lu, P., & Hu, C. X. (2007). Biotechnology Advances, 25, 1–12.Google Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Wenya Wang
    • 1
    • 2
  • Jacob A. Englaender
    • 4
  • Peng Xu
    • 3
  • Krunal K. Mehta
    • 3
  • Jiraporn Suwan
    • 2
  • Jonathan S. Dordick
    • 3
    • 4
    • 5
    • 6
  • Fuming Zhang
    • 3
  • Qipeng Yuan
    • 1
  • Robert J. Linhardt
    • 2
    • 3
    • 4
    • 5
    • 7
  • Mattheos Koffas
    • 3
    • 7
  1. 1.College of Life Science and TechnologyBeijing University of Chemical TechnologyBeijingChina
  2. 2.Department of Chemistry and Chemical BiologyRensselaer Polytechnic InstituteTroyUSA
  3. 3.Department of Chemical and Biological EngineeringRensselaer Polytechnic InstituteTroyUSA
  4. 4.Department of BiologyRensselaer Polytechnic InstituteTroyUSA
  5. 5.Department of Biomedical EngineeringRensselaer Polytechnic InstituteTroyUSA
  6. 6.Department of Materials Science and Engineering Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic InstituteTroyUSA
  7. 7.Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic InstituteTroyUSA

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