PREPARATIVE SEPARATION OF COMPONENTS OF ANTIFUNGAL ANTIBIOTIC ANTIMYCIN BY USING OF HIGH-SPEED COUNTER-CURRENT CHROMATOGRAPHY

Valery V. Belakhov, Schulich Faculty of Chemistry, Technion – Israel Institute of Technology, Haifa, Israel (chvalery@technion.ac.il)
Irina V. Boikova, Microbiological Plant Protection Laboratory, All-russian Institute Of Plant Protection, Saint -petersburg, Russia
Vera A. Kolodyaznaya, Department Of Biotechnology, Saint-petersburg State Chemical Pharmaceutical University, Saint -petersburg, Russia

Antimycin A components belong to a family of antifungal antibiotics, which containing a common nine-membered dilactone ring in their structure. At present time more than 20 antimycin A components have been prepared as secondary metabolites from various strains of Streptomyces, and antimycin A_1b is a major component of the antimycin A mixture. Antimycin A is used in agriculture due to its powerful antifungal action in the relation to various species of fungi, low toxicity and high stability. In view of this, the search of novel high effective derivatives on a base of antimycin A_1b with more wide specrtrum of biological action is a very urgent target.

 

It is known that various chromatographic methods have been applied for the separation of antimycin A components during last decades. The high-speed counter-current chromatography (HSCCC) is now accepted as an efficient preparative technique, and widely used for separation and purification of various natural and synthetic products.

 

In this communication, we report the results of an application of HSCCC for the separation of antimycin A components. The mixture of antimycin A components (sample from Sigma A8674) were separated by HSCCC with a two-phase solvent system composed of n-heptan-ethyl acetate-ethanol-water (7:4:3:0.5). The fractions were analyzed by high-performance liquid chromatography and electrospray mass spectrometry. Four components of antimycin A, and in particular antimycin A_1b, were successfully prepared, while the purity of the components of antimycin A was in range of 95.7-97.5%. The structure and molecular weights of the components of antimycin A were confimed  by ¹H and ¹³C NMR-, IR- and UV-spectroscopy and mass-spectroscopy (electrospray ionization), correspondingly.