Chemoenzymatic quantification for monitoring unpurified polysaccharide in rich medium.

摘要: The heparosan polysaccharide serves as the starting carbon backbone for chemoenzymatic synthesis of heparin, a widely used clinical anticoagulant drug. previous quantification methods rely on time-consuming purification or expensive instruments not readily available many labs. Here, approach is developed to monitor production in rich medium without purification. After removing interfering small molecules by ultrafiltration, was decomposed into oligosaccharides using heparin lyase III. were separated from large ultrafiltration and quantitatively determined anthrone-sulfuric acid assay spectrophotometer. Based different substrate specificity lyases, study showed that concentration mixture discriminatively two-step assay. Furthermore, observed be more reliable than phenol-sulfuric under these conditions. Besides may adapted quantify other types polysaccharides if specific lyases available.