Applied Biochemistry and Biotechnology

, Volume 171, Issue 4, pp 954–962

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

DOI: 10.1007/s12010-013-0415-8

Cite this article as:
Wang, W., Englaender, J.A., Xu, P. et al. Appl Biochem Biotechnol (2013) 171: 954. doi:10.1007/s12010-013-0415-8

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-SulfotransferaseBioengineered heparinEndotoxin freeBacillus subtilisBacillus megateriumNonanimal sources

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