Preparation of highly purified timosaponin AIII from rhizoma anemarrhenae through an enzymatic method combined with preparative liquid chromatography

Abstract

Timosaponin AIII (TAIII) exhibits extensive pharmacological activities and has been reported as a potent antitumour agent for various human cancers. In the present study, a potential industrial process for producing TAIII that involves biotransformation directly in the crude extract liquid of rhizoma anemarrhenae (RA) was developed. β-D-glycosidase was used to transform timosaponin BII (TBII) into TAIII, and monofactor experiments were conducted to optimise the enzymolysis conditions. In addition, AB-8 macroporous resin column chromatography, preparative liquid chromatography, and crystallisation technique were applied for yielding TAIII crystals with a purity > 97%. Approximately, 7 g of TAIII with a high purity of > 97% was obtained from 1 kg of RA through this five-step preparation method, which can be used to produce TAIII on a large scale.

Keywords:

• β-D-glycosidase
• enzymolysis
• preparative liquid chromatography
• timosaponin AIII

Funding

This work was supported by the programmes of the National Natural Science Foundation of China [grant number 81173561], [grant number 81274200], [grant number 81403175]; Youth Talent Sail Plan from Shanghai Committee of Science and Technology [grant number 14YF1411300]; Project from Shanghai Municipal Commission of Health and Family planning [grant number 20134Y053]; Project from Shanghai Committee of Science and Technology [grant number 13401900301].