Abstract
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.
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The authors are grateful to the National Institutes of Health for supporting this research with grants GM38060 and HL096972.
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Ly, M., Wang, Z., Laremore, T.N. et al. Analysis of E. coli K5 capsular polysaccharide heparosan. Anal Bioanal Chem 399, 737–745 (2011). https://doi.org/10.1007/s00216-010-3679-7
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DOI: https://doi.org/10.1007/s00216-010-3679-7