Toward a biotechnological heparin through combined chemical and enzymatic modification of the Escherichia coli K5 polysaccharide.
A process to generate glycosaminoglycans with heparin- and heparan sulfate-like sequences from the Escherichia coli K5 capsular polysaccharide is described. This polymer has the same structure as N-acetylheparosan, the precursor in heparin/ heparan sulfate biosynthesis. The process involves chemical N-deacetylation and N-sulfation, enzymatic conversion of up to 60% of the D-glucuronic acid to L-iduronic acid residues, and chemical O-sulfation. Because direct sulfation afforded unwanted 3-O-sulfated (instead of 2-O-sulfated) iduronic acid residues, a strategy involving graded solvolytic desulfation of chemically oversulfated C5-epimerized sulfaminoheparosans was assessed using persulfated heparin and heparan sulfate as model compounds. The O-desulfation process was shown to increase the anti-factor Xa activity of oversulfated heparin.
Naggi
A
A
G. Ronzoni Institute for Chemical and Biochemical Research, Milan, Italy.
Torri
G
G
Casu
B
B
Oreste
P
P
Zoppetti
G
G
Li
J P
JP
Lindahl
U
U
D000818:Animals; D016667:Bacterial Capsules; D001709:Biotechnology; D004926:Escherichia coli / Q000737:chemistry*; D015951:Factor Xa / Q000378:metabolism; D065427:Factor Xa Inhibitors; D006493:Heparin / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D013431:Sulfates / Q000737:chemistry
D065427:Factor Xa Inhibitors; D011135:Polysaccharides, Bacterial; D013431:Sulfates; C050673:capsular polysaccharide K5; D006493:Heparin; D015951:Factor Xa