Simple Method for the Preparation of Teadenols A and B by a Combined Process of Submerged Culture with Aspergillus sp. and Chromatographic Separation

Abstract

Methods for the production and isolation of the tea components teadenols A and B were developed because of their potential anti-metabolic syndrome activity. Aspergillus sp. (FARM AP-21280) was inoculated to a modified Czapek-Dox medium containing (-)-epigallocatechin 3-O-gallate (EGCG) and (-)-gallocatechin 3-O-gallate (GCG) and incubated at 25°C. The maximum contents of the teadenols were observed at 19 days, and the contents increased depending on the initial EGCG concentration. The addition of green tea powder to the medium enhanced the production of teadenols. In the modified Czapek-Dox medium containing 5.0% EGCG and 1.0% green tea powder, teadenols A and B were efficiently produced at 3.22 ± 0.23 and 4.49 ± 0.38 mg/mL, respectively. The teadenols were purified by sequential chromatography on Sephadex LH-20 and InertSustain™ C18 columns. After purification by preparative HPLC, 0.27 g teadenol A (29.7% recovery) and 0.45 g teadenol B (42.9% recovery) were obtained from the 970 mL culture filtrate.