E. coli K5 fermentation and the preparation of heparosan,a bioengineered heparin precursor |
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| Authors: | Zhenyu Wang Mellisa Ly Fuming Zhang Weihong Zhong Amy Suen Anne Marie Hickey Jonathan S Dordick Robert J Linhardt |
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| Institution: | 1. Department of Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York 12180;2. telephone: 518‐355‐4062;3. fax: 518‐276‐2207;4. Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York 12180;5. Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York 12180;6. Department of Biotechnology, College of Biological & Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, PR China |
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| Abstract: | Heparosan is an acidic polysaccharide natural product, which serves as the critical precursor in heparin biosynthesis and in the chemoenzymatic synthesis of bioengineered heparin. Heparosan is also the capsular polysaccharide of Escherichia coli K5 strain. The current study was focused on the examination of the fermentation of E. coli K5 with the goal of producing heparosan in high yield and volumetric productivity. The structure and molecular weight properties of this bacterial heparosan were determined using polyacrylamide gel electrophoresis (PAGE) and Fourier transform mass spectrometry. Fermentation of E. coli K5 in a defined medium using exponential fed‐batch glucose addition with oxygen enrichment afforded heparosan at 15 g/L having a number average molecular weight of 58,000 Da and a weight average molecular weight of 84,000 Da. Biotechnol. Bioeng. 2010;107: 964–973. © 2010 Wiley Periodicals, Inc. |
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| Keywords: | fermentation E coli capsule polysaccharide heparosan heparin |
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