Isolation and purification of macrocyclic components from Penicillium fermentation broth by high-speed counter-current chromatography.
In this paper, high-speed counter-current chromatography (HSCCC), assisted with ESI-MS, was first successfully applied to the preparative separation of three macrolide antibiotics, brefeldin A (12.6 mg, 99.0%), 7'-O-formylbrefeldin A (6.5 mg, 95.0%) and 7'-O-acetylbrefeldin A (5.0 mg, 92.3%) from the crude extract of the microbe Penicillium SHZK-15. Considering the chemical nature and partition coefficient (K) values of the three target compounds, a two-step HSCCC isolation protocol was developed in order to obtain products with high purity. In the two-step method, the crude ethyl acetate extract was first fractionated and resulted in two peak fractions by HSCCC using solvent system n-hexane/ethyl acetate/methanol/water (HEMWat) (3:7:5:5 v/v/v/v), then purified using solvent systems HEMWat (3:5:3:5 v/v/v/v) and HEMWat (7:3:5:5 v/v/v/v) for each fraction. The purities and structures of the isolated compounds were determined by HPLC, X-ray crystallography, ESI-MS and NMR. The results demonstrated that HSCCC is a fast and efficient technique for systematic isolation of bioactive compounds from the microbes.
Gao
Xiang
X
Department of Chemistry and The Key Laboratory for Chemical Biology of Fujian Province, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, PR China.
Zhuang
Rongqiang
R
Guo
Jiannan
J
Bao
Jian
J
Fang
Meijuan
M
Liu
Yan
Y
Xu
Pengxiang
P
Zhao
Yufen
Y
D000085:Acetates / Q000737:chemistry; D020126:Brefeldin A / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution / Q000379:methods*; D018360:Crystallography, X-Ray; D017077:Culture Media, Conditioned / Q000737:chemistry; D005285:Fermentation; D047028:Macrocyclic Compounds / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D010407:Penicillium / Q000378:metabolism*; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet
D000085:Acetates; D017077:Culture Media, Conditioned; D047028:Macrocyclic Compounds; D012997:Solvents; C539298:brefeldin A formylate; D020126:Brefeldin A; C007650:ethyl acetate