Recovery and purification of thuringiensin from the fermentation broth of Bacillus thuringiensis

Abstract

Thuringiensin is a heat stable β-exotoxin from Bacillus thuringiensis with a great potential for replacing the traditional chemical pesticides. A process using micellar-enhanced ultrafitration method to recover thuringiensin was significantly improved by the use of a spiral-wound membrane, which could be operated at a low transmembrane pressure drop. This method was performed by adding a surfactant cetylpyridinium chloride (CPC) into the fermentation broth. After the surfactant-thuringiensin conjugates were formed, the broth then passed through the ultrafiltration membrane and the retentate was collected. The results indicated the optimal concentration of CPC for producing a maximal recovery up to 99.3% is 4%. For purification, the centrifuged broth was further filtered by a membrane filter. The filtered solution then was mixed with 50% of activated carbon. The supernatant then was injected into a preparative HPLC. The eluate was collected during thuringiensin peak formation. This eluate was then concentrated by vacuum evaporation and dialysis using an electrodialyzer to remove the excess salts. The dialyzed solution was then crystallized by lyophilization. The purity of the thuringiensin crystal was identified by HPLC, capillary electrophoresis, and mass spectrometry.