Method for removing bacterial endotoxin from Gram-negative polysaccharides

摘要

PURPOSE: To remove the bacterial endotoxin from a Gram-negative polysaccharide without incurring a substantial loss of the polysaccharide by pre-purifying a Gram-negative bacteria fermentation product and then carrying out selective alcohol fractionation and resin removal treatment. ;CONSTITUTION: An endotoxin is removed from a Gram-negative bacteria fermentation product by the following steps (a)-(e). (a) Gram-negative bacteria are propagated in a fermentation broth to release polysaccharide in the broth and an alcohol is added to the broth to remove impurities by precipitation; (b) the remaining high mol.wt. substances are isolated and re-dissolved is phenol to extract other impurities; (c) the remaining high mol.wt. substances are centrifuged and re-dissolved in a divalent counter ion solution; (d) an alcohol is added to the solution and the solution is cooled and then, an alcohol is gradually increasingly added to precipitate a lipopolysaccharide/polysaccharide; and (e) the lipopolysaccharide/polysaccharide-containing substance formed by the addition of the alcohol is mixed with a nonionic resin, a surfactant and a chelating agent to remove the lipopolysaccharide. As the polysaccharide, polyribosylribitol phosphate can be mentioned.;COPYRIGHT: (C)1991,JPO