BORDERS DB, FRANCIS ND, SIMON RJ, FANTINI AA; Interscience Conference on Antimicrobial Agents and Chemotherapy (41st : 2001 : Chicago, Ill.).

Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 2001 Dec 16-19; 41: abstract no. F-1156.

BioSource Pharm, Spring Valley, NY

BACKGROUND: The original procedures developed for the isolation and purification of the laspartomycin complex from fermentation broths involved a number of extraction and chromatography steps to obtain material free of pigments and other impurities. Purification of other lipopeptide antibiotic complexes such as amphomycin and aspartocin, and the lipodepsipeptide daptomycin are complicated by similar problems. METHODS: Fermentations of Streptomyces viridochromogenes ssp. komabensis gave the laspartomycin complex at approximately 500 microg/ml. Laspartomycin and other related lipopeptide antibiotics were found to form calcium chelates. The chelates are stable under basic conditions and can be extracted into organic solvents such as 1-butanol. Under acidic conditions the chelates are degraded. They can then be back-extracted into water at near neutral pH conditions. RESULTS: By a combination of chelate and standard extraction procedures with subsequent freeze drying of product, relatively pure samples of the partial sodium salt form of the laspartomycin complex were obtained as white powders. HPLC analysis showed that this complex contained 70-80% of a major component bearing a C[15] side chain, C[57]H[89]N[12]O[19]Na, HRFABMS m/z (M+Na)[+] calc.1269.6343, obsd. 1269.6289. The salt form of the complex was converted to the free acid, a white powder with the major component C[57]H[90]N[12]O[19], FABMS m/z (M+H)[+] calc. 1248, obsd. (M+H)[+] 1248, (M+Na)[+] 1270, (M+K)[+] 1286.The amphomycin, aspartocin and daptomycin complexes were also successfully purified by the chelate procedure to obtain the corresponding purified complexes. CONCLUSION: The new chelate purification process for lipopeptide antibiotics provides a rapid and direct method of purification of acidic lipopeptide antibiotics from fermentation broths.

Publication Types:

• Meeting Abstracts

Keywords:

• Anti-Bacterial Agents
• Chromatography, High Pressure Liquid
• Daptomycin
• Fermentation
• Hydrogen-Ion Concentration
• Oligopeptides
• Spectrometry, Mass, Fast Atom Bombardment
• Streptomyces
• amphomycin
• methods

Other ID:

• GWAIDS0030344

UI: 102269981

From Meeting Abstracts