Sulfonation and anticoagulant activity of botryosphaeran from Botryosphaeria rhodina MAMB-05 grown on fructose

Botryosphaeran (EPSFRU), an exopolysaccharide of the β-(1→3,1→6)-d-glucan type with 31% branching at C-6, is produced by the fungus Botryosphaeria rhodina MAMB-05 when grown on fructose as carbon source. Botryosphaeran was derivatized by sulfonation to induce anticoagulant activity. The effectiveness of the sulfonation reaction by chlorosulfonic acid in pyridine was monitored by the degree of substitution and FT-IR analysis of the sulfonated EPSFRU (once sulfonated, EPSFRU SULF; and re-sulfonated, EPSFRU RESULF). Activated partial thromboplastin time (APTT) and thrombin time (TT) tests of EPSFRU RESULF indicated significant in vitro anticoagulant activity that was dose-dependent. EPSFRU did not inhibit any of the coagulation tests.