N-acetyl-heparosan lyase of Escherichia coli K5: gene cloning and expression. |
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| Authors: | 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 Salom |
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| Affiliation: | 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 Salomé |
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| Abstract: | 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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