title: Analysis of E. coli K5 capsular polysaccharide heparosan.
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 (M(N)), weight-averaged MWs (M(W)), and MW ranges of 3,000 to 150,000 Da.
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, 110 Eight Street, Troy, NY 12180, USA.
authors.forename: Mellisa
authors.initials: M
authors.lastname: Ly
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Wang
authors.forename: Tatiana N
authors.initials: TN
authors.lastname: Laremore
authors.forename: Fuming
authors.initials: F
authors.lastname: Zhang
authors.forename: Weihong
authors.initials: W
authors.lastname: Zhong
authors.forename: Dennis
authors.initials: D
authors.lastname: Pu
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Zagorevski
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Dordick
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
mesh_terms: D000925:Anticoagulants / Q000737:chemistry* / Q000378:metabolism; D016667:Bacterial Capsules / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D005285:Fermentation; D005583:Fourier Analysis; D008970:Molecular Weight; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000925:Anticoagulants; C050673:capsular polysaccharide K5
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