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 language | English |
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Pages (from-to) | 954-962 |
Number of pages | 9 |
Journal | Applied Biochemistry and Biotechnology |
Volume | 171 |
Issue number | 4 |
DOIs | |
State | Published - Oct 2013 |
Externally published | Yes |
Keywords
- 3-O-Sulfotransferase
- Bacillus megaterium
- Bacillus subtilis
- Bioengineered heparin
- Endotoxin free
- Nonanimal sources