| Autor: |
Pan, C. H., Chan, G., Nisbet, L. J., Polansky, M. J., Vogt-Speth, S., Sitrin, R. D. |
| Zdroj: |
Journal of Industrial Microbiology and Biotechnology; February 1987, Vol. 1 Issue: 5 p303-310, 8p |
| Abstrakt: |
Summary A gradient analytical HPLC system was developed to assay titers of the three major components of the aridicin (Ardacin) complex produced byKibdelosporangium aridum (SK&F AAD-216). The separation was performed on a Beckman Ultrasphere column using a gradient of acetonitrile (26–43%) in 0.1 M pH 3.2 phosphate buffer with UV detection at 220 nm. The gradient system was necessary to analyze all three major factors within a reasonable recycle time (14 min) without interference by front eluting impurities. The assay was linear from 12 to 200 µg/ml (multipleR2=0.998), with a standard deviation for retention time of 1.4%. A SepPAK isolation scheme was developed to assay samples in complex matrices such as fermentation broths. Using this assay as a monitor, fermentation medium optimization increased the total titers of the three factors from approximately 5 µg/ml to over 200 µg/ml. The optimal medium contained glucose, beet molasses and methyl oleate. The latter substrate was particularly effective in enhancingproducation 10-fold, presumably by enhancing the supply of acetly-CoA. This is a biosynthetic precursor of both dihydroxyphenylglycine, present in the nucleus, and the acyl side chains present on the amino-glucuronic acids. |
| Databáze: |
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