High Cell Density Cultivation of Recombinant Escherichia coli Strains Expressing 2-O-Sulfotransferase and C5-Epimerase for the Production of Bioengineered Heparin

Jianhua Zhang, Matt Suflita, Guoyun Li, Weihong Zhong, Lingyun Li, Jonathan S. Dordick, Robert J. Linhardt, Fuming Zhang

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Bioengineered heparin is being investigated as a potential substitute for the animal-sourced anticoagulant drug. One step in the current process to prepare bioengineered heparin involves the conversion of N-sulfo heparosan, rich in → 4)GlcNS(1 → 4) GlcA(1 → sequences (where S is sulfo, GlcN is α-d-glucosamine, and GlcA is β-d-glucuronic acid), to a critical intermediate, rich in → 4)GlcNS(1 → 4) IdoA2S(1 → sequences (where S is sulfo and IdoA is α-l-iduronic acid), using 2-O-sulfotransferase (2-OST) and C5 epimerase (C5-epi). Until now, these heparan sulfate biosynthetic enzymes have been expressed in Escherichia coli grown in shake flask culture as fusion proteins. The current study is focused on the high cell density fed-batch cultivation of recombinant E. coli strains expressing both enzymes. We report the high productivity expression of active 2-OST and C5-epi enzymes of 6.0 and 2.2 mg/g dry cell weight, respectively.

Original languageEnglish
Pages (from-to)2986-2995
Number of pages10
JournalApplied Biochemistry and Biotechnology
Volume175
Issue number6
DOIs
StatePublished - Mar 2015
Externally publishedYes

Keywords

  • Epimerase
  • Fermentation
  • Heparin
  • Recombinant enzymes
  • Sulfotransferase

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