N-acetyl-heparosan lyase of Escherichia coli K5: gene cloning and expression.
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.
Legoux
R
R
Department of Microbiology, Sanofi-Recherche, Labege, France. richard.legoux@tls1.elfsanofi.fr
Lelong
P
P
Jourde
C
C
Feuillerat
C
C
Capdevielle
J
J
Sure
V
V
Ferran
E
E
Kaghad
M
M
Delpech
B
B
Shire
D
D
Ferrara
P
P
Loison
G
G
Salomé
M
M
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
D003470:Culture Media; D004269:DNA, Bacterial; D011135:Polysaccharides, Bacterial; D011133:Polysaccharide-Lyases