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Analysis of E. coli K5 capsular polysaccharide heparosan
- Ly, Mellisa, Wang, Zhenyu, Laremore, Tatiana N., Zhang, Fuming, Zhong, Weihong, Pu, Dennis, Zagorevski, Dmitri V., Dordick, Jonathan S., Linhardt, Robert J.
- Analytical and bioanalytical chemistry 2011 v.399 no.2 pp. 737-745
- Escherichia coli, anticoagulants, fermentation, fractionation, heparin, ionization, mass spectrometry, molecular weight, polyacrylamide gel electrophoresis, swine
- Heparosan is the key precursor for the preparation of bioengineered heparin, a potential replacement for porcine intestinal heparin, an important anticoagulant drug. The molecular weight (MW) distribution of heparosan produced by the fermentation of E. coli K5 was investigated. Large-slab isocratic and mini-slab gradient polyacrylamide gel electrophoresis (PAGE) were used to analyze the MW and polydispersity of heparosan. A preparative method that allowed fractionation by continuous-elution PAGE was used to obtain heparosan MW standards. The MWs of the heparosan standards were determined by electrospray ionization Fourier-transform mass spectrometry (ESI-FT-MS). A ladder of the standards was then used to determine the MW properties of polydisperse heparosan samples. Unbleached and bleached heparosan produced by fermentation of E. coli K5 had similar number-averaged MWs (MN), weight-averaged MWs (MW), and MW ranges of 3,000 to 150,000 Da.