Liquid chromatographic assay of microbially derived phloroglucinol antibiotics for establishing the biosynthetic route to production, and the factors affecting their regulation
✍ Scribed by P. Shanahan; J.D. Glennon; J.J. Crowley; D.F. Donnelly; F. O'Gara
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 656 KB
- Volume
- 272
- Category
- Article
- ISSN
- 0003-2670
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✦ Synopsis
A preparatwe chromatographlc Isolation method mvolvmg thm-layer and hqmd chromatography (LC) 1s described for the nucrotually denved antdxotic monoacetylphloroglucmol (MAPG) Isolation of MAPG along with the previously described 2,4_dlacetylphloroglucmol (DAPG) confIrms the productIon of two antmcrotual metabohtes from Pseudo~~nus sp stram F113 A gradient LC assay was developed for the determmation of these compounds m growth culture media A sample pretreatment procedure mvolvmg solid phase extraction on octadecylsdxa IS utrhsed poor to sample lqlection onto the LC column This assay 1s used to monitor production of the two antlblotic metabohtes from the strain of PseudomoMs A range of nitrogen sources m the growth culture medium was tested for their ablllty to at&t antibiotic production A carbon source, such as galactose promotes high yields of antlblotlcs when the growth medium contams a mtrogen source m the form of ammomum ions, and produces low yields when the mtrogen source LS added as nitrate ions The LC assay 1s further applied to monitor the enzymatic acetylation of MAPG to DAPG, through an enzymatic actMty named MAPG acetyltransferase which provides the final step m the elucidation of the biosynthesis of DAPG