Expression of low endotoxin 3-O-sulfotransferase in Bacillus subtilis

Wang, Wenya
Englaender, Jacob A.
Xu, Peng
Mehta, Krunal K.
Suwan, Jiraporn
Dordick, Jonathan S.
Zhang, Fuming
Yuan, Qipeng
Linhardt, Robert J.
Koffas, Mattheos
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Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Expression of low endotoxin 3-O-sulfotransferase in Bacillus subtilis, W. Wang, P. Xu, J. Dordick, F. Zhang, Q. Yuan, M. Koffas, R.J. Linhardt, Applied Biochemistry and Biotechnology, 171, 954–962, 2013.
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 10(4)-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.
Applied Biochemistry and Biotechnology, 171, 954–962
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The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
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Rensselaer Polytechnic Institute, Troy, NY
Applied Biochemistry and Biotechnology