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

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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.

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Acknowledgments

This work was supported by grants funded by the National Institutes of Health HL101721 and HL096972 (RJL), the Bioengineered Heparin Consortium, and 863 Hi-Tech Research and Development Program of the People’s Republic of China (project no. 2012AA022300). The authors would also like to thank Dr. Sui-Lam Wong (University of Calgary), Dr. Xiaozhou Zhang (Virginia Tech), and Dr. Cynthia Collins (Rensselaer Polytechnic Institute) for supplying bacterial strains and plasmids.

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Authors and Affiliations

  1. College of Life Science and Technology, Beijing University of Chemical Technology, Box 75, Beijing, 100029, China

    Wenya Wang & Qipeng Yuan

  2. Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA

    Wenya Wang, Jiraporn Suwan & Robert J. Linhardt

  3. Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA

    Peng Xu, Krunal K. Mehta, Jonathan S. Dordick, Fuming Zhang, Robert J. Linhardt & Mattheos Koffas

  4. Department of Biology, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA

    Jacob A. Englaender, Jonathan S. Dordick & Robert J. Linhardt

  5. Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA

    Jonathan S. Dordick & Robert J. Linhardt

  6. Department of Materials Science and Engineering Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA

    Jonathan S. Dordick

  7. Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Biotech 4005, 110 8th Street, Troy, NY, 12180, USA

    Robert J. Linhardt & Mattheos Koffas

Authors
  1. Wenya Wang
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  2. Jacob A. Englaender
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  3. Peng Xu
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  4. Krunal K. Mehta
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  5. Jiraporn Suwan
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  6. Jonathan S. Dordick
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  7. Fuming Zhang
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  8. Qipeng Yuan
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  9. Robert J. Linhardt
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  10. Mattheos Koffas
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Corresponding authors

Correspondence to Robert J. Linhardt or Mattheos Koffas.

Additional information

Wenya Wang and Jacob A. Englaender equally contributed to this work.

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Wang, W., Englaender, J.A., Xu, P. et al. Expression of Low Endotoxin 3-O-Sulfotransferase in Bacillus subtilis and Bacillus megaterium . Appl Biochem Biotechnol 171, 954–962 (2013). https://doi.org/10.1007/s12010-013-0415-8

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  • DOI: https://doi.org/10.1007/s12010-013-0415-8

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