Article

Applied Biochemistry and Biotechnology

, Volume 171, Issue 4, pp 954-962

First online:

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

  • Wenya WangAffiliated withCollege of Life Science and Technology, Beijing University of Chemical TechnologyDepartment of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute
  • , Jacob A. EnglaenderAffiliated withDepartment of Biology, Rensselaer Polytechnic Institute
  • , Peng XuAffiliated withDepartment of Chemical and Biological Engineering, Rensselaer Polytechnic Institute
  • , Krunal K. MehtaAffiliated withDepartment of Chemical and Biological Engineering, Rensselaer Polytechnic Institute
  • , Jiraporn SuwanAffiliated withDepartment of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute
  • , Jonathan S. DordickAffiliated withDepartment of Chemical and Biological Engineering, Rensselaer Polytechnic InstituteDepartment of Biology, Rensselaer Polytechnic InstituteDepartment of Biomedical Engineering, Rensselaer Polytechnic InstituteDepartment of Materials Science and Engineering Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute
  • , Fuming ZhangAffiliated withDepartment of Chemical and Biological Engineering, Rensselaer Polytechnic Institute
  • , Qipeng YuanAffiliated withCollege of Life Science and Technology, Beijing University of Chemical Technology
  • , Robert J. LinhardtAffiliated withDepartment of Chemistry and Chemical Biology, Rensselaer Polytechnic InstituteDepartment of Chemical and Biological Engineering, Rensselaer Polytechnic InstituteDepartment of Biology, Rensselaer Polytechnic InstituteDepartment of Biomedical Engineering, Rensselaer Polytechnic InstituteCenter for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute Email author 
    • , Mattheos KoffasAffiliated withDepartment of Chemical and Biological Engineering, Rensselaer Polytechnic InstituteCenter for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute Email author 

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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.

Keywords

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