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

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 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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