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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.

SUBMITTER: Ly M 

PROVIDER: S-EPMC3390146 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature

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Analysis of E. coli K5 capsular polysaccharide heparosan.

Ly Mellisa M   Wang Zhenyu Z   Laremore Tatiana N TN   Zhang Fuming F   Zhong Weihong W   Pu Dennis D   Zagorevski Dmitri V DV   Dordick Jonathan S JS   Linhardt Robert J RJ  

Analytical and bioanalytical chemistry 20100421 2


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  ...[more]

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