Analysis of E. coli K5 capsular polysaccharide heparosan.

Analysis of E. coli K5 capsular polysaccharide heparosan.
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DOI:
10.1007/s00216-010-3679-7
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发表时间:
2011-01
影响因子:
4.3
通讯作者:
Linhardt, Robert J.
Linhardt, Robert J.
中科院分区:
化学2区
文献类型:
--
作者:
Ly, Mellisa;Wang, Zhenyu;Laremore, Tatiana N.;Zhang, Fuming;Zhong, Weihong;Pu, Dennis;Zagorevski, Dmitri V.;Dordick, Jonathan S.;Linhardt, Robert J.

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肝素原是制备生物工程肝素的关键前体,是重要抗凝药物猪肠肝素的潜在替代品。对E. coli K5中进行了初步研究。采用大平板等度和小平板梯度聚丙烯酰胺凝胶电泳(PAGE)分析肝素原的分子量和多分散性。使用允许通过连续洗脱PAGE分级分离的制备方法来获得肝素原MW标准品。通过电喷雾电离傅里叶变换质谱法(ESI-FT-MS)测定肝素原标准品的MW。然后使用标准品的阶梯来确定多分散肝素原样品的MW性质。通过E. coli K5具有相似的数均分子量(MN)、重均分子量(MW)和3,000至150,000 Da的MW范围。
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.
DOI: 10.1021/ja8026345
发表时间: 2008-10-01
影响因子: 15
作者:
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