title: N-acetyl-heparosan lyase of Escherichia coli K5: gene cloning and expression.
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.
authors.affiliation: Department of Microbiology, Sanofi-Recherche, Labege, France. richard.legoux@tls1.elfsanofi.fr
authors.forename: R
authors.initials: R
authors.lastname: Legoux
authors.forename: P
authors.initials: P
authors.lastname: Lelong
authors.forename: C
authors.initials: C
authors.lastname: Jourde
authors.forename: C
authors.initials: C
authors.lastname: Feuillerat
authors.forename: J
authors.initials: J
authors.lastname: Capdevielle
authors.forename: V
authors.initials: V
authors.lastname: Sure
authors.forename: E
authors.initials: E
authors.lastname: Ferran
authors.forename: M
authors.initials: M
authors.lastname: Kaghad
authors.forename: B
authors.initials: B
authors.lastname: Delpech
authors.forename: D
authors.initials: D
authors.lastname: Shire
authors.forename: P
authors.initials: P
authors.lastname: Ferrara
authors.forename: G
authors.initials: G
authors.lastname: Loison
authors.forename: M
authors.initials: M
authors.lastname: Salomé
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003470:Culture Media; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000235:genetics* / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000378:metabolism*; D013045:Species Specificity; D014170:Transformation, Genetic
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D011135:Polysaccharides, Bacterial; D011133:Polysaccharide-Lyases