|
|||||
|
|
Expression of Low Endotoxin 3-O-Sulfotransferase in Bacillus subtilis and Bacillus megaterium |
|
| Authors: | Wenya Wang Jacob A. Englaender Peng Xu Krunal K. Mehta Jiraporn Suwan Jonathan S. Dordick Fuming Zhang Qipeng Yuan Robert J. Linhardt Mattheos Koffas |
| Affiliation: | 1. College of Life Science and Technology, Beijing University of Chemical Technology, Box 75, Beijing, 100029, China 2. Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA 4. Department of Biology, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA 3. Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA 5. Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA 6. Department of Materials Science and Engineering Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, 12180, USA 7. Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Biotech 4005, 110 8th Street, Troy, NY, 12180, USA |
| 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: | |
| 本文献已被 SpringerLink 等数据库收录！ | |