HPLC-fluorescence detection method for determination of key intermediates of the lincomycin biosynthesis in fermentation broth
Language English Country Germany Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Amides analysis MeSH
- Fermentation * MeSH
- Fluorescence * MeSH
- Lincomycin analogs & derivatives biosynthesis MeSH
- Molecular Structure MeSH
- Proline analogs & derivatives analysis MeSH
- Reproducibility of Results MeSH
- Streptomyces metabolism MeSH
- Sulfhydryl Compounds analysis MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 4-n-propyl-L-proline MeSH Browser
- Amides MeSH
- Lincomycin MeSH
- Proline MeSH
- Sulfhydryl Compounds MeSH
The biosynthetic pathway of the clinically important antibiotic lincomycin is not known in details. The precise knowledge of the lincomycin biosynthesis is a prerequisite for generation of improved derivatives by means of combinatorial genetics. Methods allowing determination of the key intermediates are very important tools of the pathway investigation. Two new high-performance liquid chromatography methods with fluorescence detection for determination of lincomycin precursors in fermentation broth of Streptomyces lincolnensis and its lincomycin nonproducing mutants were developed. The first one enables simultaneous analysis of methylthiolincosamide (MTL) and N-demethyllincomycin (NDL), whereas the second one is suitable for 4-propyl-L-proline (PPL) assay. Both methods are based on the pre-column derivatization: MTL and NDL with 4-chloro-7-nitrobenzofurazan; PPL with o-phthaldialdehyde. The methods were validated with lower limit of quantification values of 2.50, 3.75, and 3.75 microg ml(-1) for MTL, NDL, and PPL, respectively. The inter- and intra-day accuracies and precisions were all within 12%. Stability of oxidized and derivatized analytes was investigated.
References provided by Crossref.org
Mutasynthesis of lincomycin derivatives with activity against drug-resistant staphylococci