J. Bacteriol., 12 1996, 7260-7264, Vol 178, No. 24
Copyright © 1996, American Society for Microbiology

N-acetyl-heparosan lyase of Escherichia coli K5: gene cloning and expression

R Legoux, P Lelong, C Jourde, C Feuillerat, J Capdevielle, V Sure, E Ferran, M Kaghad, B Delpech, D Shire, P Ferrara, G Loison and M Salome
Department of Microbiology, Sanofi-Recherche, Labege, France. richard.legoux@tls1.elfsanofi.fr

The structure of the capsular polysaccharide of Escherichia coli K5 is identical to that of N-acetyl-heparosan, a nonsulfated precursor of heparin, which makes this E. coli antigen an attractive starting point for the chemical synthesis of analogs of low-molecular-weight heparin. This polysaccharide is synthesized as a high-molecular-weight molecule that can be depolymerized by an enzyme displaying endo-beta-eliminase activity. The eliminase-encoding gene, designated elmA, has been cloned from E. coli K5 by expression in E. coli K-12. The K-12 genome is devoid of the elmA sequence. The elmA gene product is 820 amino acids long. Active recombinant eliminase is produced by K-12 cells in both cell-bound and secreted forms. Deletion analyses have shown that the C terminus and the N terminus are required for activity and secretion, respectively.

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