Preparative isolation and purification of anti-tumor agent ansamitocin P-3 from fermentation broth of Actinosynnema pretiosum using high-performance counter-current chromatography.
Ansamitocin P-3 is a potent anti-tumor maytansinoid found in Actinosynnema pretiosum. However, due to the complexity of the fermentation broth of Actinomycete, how to effectively separate ansamitocin P-3 is still a challenge. In this study, both analytical and preparative high-performance counter-current chromatography were successfully used to separate and purify ansamitocin P-3 from fermentation broth. A total of 28.8 mg ansamitocin P-3 with purity of 98.4% was separated from 160 mg crude sample of fermentation broth in less than 80 min with the two-phase solvent system of hexane-ethyl acetate-methanol-water (0.6:1:0.6:1, v/v/v/v). The purity and structural identification were determined by HPLC, (1)H NMR, (13)C NMR and mass spectroscopy.
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan Province, PR China.
Yuqin
Y
Yao
Zhihui
Z
Cheng
Haoyu
H
Ye
Yongmei
Y
Xie
Jing
J
He
Minghai
M
Tang
Tao
T
Shen
Jiangman
J
Wang
Yan
Y
Zhou
Zejun
Z
Lu
Feng
F
Luo
Lijuan
L
Chen
Luoting
L
Yu
Jin-Liang
JL
Yang
Aihua
A
Peng
Yuquan
Y
Wei
D000970:Antineoplastic Agents / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution / Q000379:methods*; D003470:Culture Media; D005285:Fermentation; D006094:Gram-Positive Bacteria / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008453:Maytansine / Q000031:analogs & derivatives* / Q000302:isolation & purification
D000970:Antineoplastic Agents; D003470:Culture Media; D008453:Maytansine; C015682:ansamitocins