ABSTRACT
A new cepham metabolite has been isolated from the filtered broth of Cephalosporium acremonium by high performance liquid chromatography (HPLC) and identified as 7 beta-(5-D-amino-adipamido)-3 beta-hydroxy-3 alpha-methyl-cepham-4 alpha-carboxylic acid (I). Pure penicillin N was prepared using HPLC in the analytical mode. When I was added in place of penicillin N as substrate for the cell-free biosynthetic of cephalosporin, no formation of deacetoxycephalosporin C (II) was observed. A synthetic cepham derivative, 7 beta-(5-D-aminoadipamido)-3-exomethylene-cepham-4 alpha-carboxylic acid (III) was also tested in the cell-free system as a possible intermediate. The compound III was shown to be an inhibitor of the ring expansion enzyme that converts penicillin N to deacetoxycephalosporin C.
Subject(s)
Acremonium/metabolism , Anti-Bacterial Agents/biosynthesis , Cephalosporins/biosynthesis , Chromatography, High Pressure Liquid , Cell-Free System , Penicillins/isolation & purificationABSTRACT
A portion of the eluent, during liquid chromatography of a fermentation broth, is applied directly to a paper strip at the same rate as the recording of the U. V. absorption. The paper is then placed on the agar inoculated with an appropriate test organism. This procedure permits direct correlation of biological activity with corresponding peaks in the U. V.
Subject(s)
Anti-Bacterial Agents/analysis , Chromatography, High Pressure Liquid/methods , Culture Media/analysis , FermentationABSTRACT
Microbonded propylamine silica with a solvent system containing acetic acid, methanol, acetonitrile and water (2:4:7.5:86.5) is suitable for an efficient separation of mixtures containing several closely related cephem derivatives. The same system with preparative columns was used for the isolation of cephalosporin C directly from the filtered broth of C. acremonium fermentation.
