Applied Biochemistry and Biotechnology

, Volume 171, Issue 4, pp 954–962

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

Authors

  • Wenya Wang
    • College of Life Science and TechnologyBeijing University of Chemical Technology
    • Department of Chemistry and Chemical BiologyRensselaer Polytechnic Institute
  • Jacob A. Englaender
    • Department of BiologyRensselaer Polytechnic Institute
  • Peng Xu
    • Department of Chemical and Biological EngineeringRensselaer Polytechnic Institute
  • Krunal K. Mehta
    • Department of Chemical and Biological EngineeringRensselaer Polytechnic Institute
  • Jiraporn Suwan
    • Department of Chemistry and Chemical BiologyRensselaer Polytechnic Institute
  • Jonathan S. Dordick
    • Department of Chemical and Biological EngineeringRensselaer Polytechnic Institute
    • Department of BiologyRensselaer Polytechnic Institute
    • Department of Biomedical EngineeringRensselaer Polytechnic Institute
    • Department of Materials Science and Engineering Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic Institute
  • Fuming Zhang
    • Department of Chemical and Biological EngineeringRensselaer Polytechnic Institute
  • Qipeng Yuan
    • College of Life Science and TechnologyBeijing University of Chemical Technology
    • Department of Chemistry and Chemical BiologyRensselaer Polytechnic Institute
    • Department of Chemical and Biological EngineeringRensselaer Polytechnic Institute
    • Department of BiologyRensselaer Polytechnic Institute
    • Department of Biomedical EngineeringRensselaer Polytechnic Institute
    • Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic Institute
    • Department of Chemical and Biological EngineeringRensselaer Polytechnic Institute
    • Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic Institute
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