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

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

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 104-10 5-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.

Original languageEnglish
Pages (from-to)954-962
Number of pages9
JournalApplied Biochemistry and Biotechnology
Volume171
Issue number4
DOIs
StatePublished - Oct 2013
Externally publishedYes

Keywords

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

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