Quantitation of heparosan with heparin lyase III and spectrophotometry |
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| Authors: | Haichan Huang Yingying Zhao Shencong Lv Weihong Zhong Fuming Zhang Robert J. Linhardt |
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| Affiliation: | 1. College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, People’s Republic of China;2. Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA;3. Department of Biology, Rensselaer Polytechnic Institute, Troy, NY 12180, USA;4. Departments of Chemistry and Chemical Biology and Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA |
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| Abstract: | Heparosan is Escherichia coli K5 capsule polysaccharide, which is the key precursor for preparing bioengineered heparin. A rapid and effective quantitative method for detecting heparosan is important in the large-scale production of heparosan. Heparin lyase III (Hep III) effectively catalyzes the heparosan depolymerization, forming unsaturated disaccharides that are measurable using a spectrophotometer at 232 nm. We report a new method for the quantitative detection of heparosan with heparin lyase III and spectrophotometry that is safer and more specific than the traditional carbazole assay. In an optimized detection system, heparosan at a minimum concentration of 0.60 g/L in fermentation broth can be detected. |
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| Keywords: | Heparosan Heparin lyase III Quantitative method Enzymatic method Escherichia coli K5 |
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