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Biosynthesis of formycin. Formation of formycin from formycin B.

1. In replacement culture with a formycin-producing strain. Steptomyces sp. MA406-A-1, exogenously added formycin B was quantitatively converted to formycin and the conversion was inhibited by adding the chromophore moiety of formycin. 2. The in vitro experiments revealed that the novel enzyme(s) catalyzing the formation and formycin from asparate and formycin B, but not from formycin B monophosphate, was present in this organism. The action of the partially purified enzyme(s) was also inhibited by the chromophore moiety of formycin, whereas the moiety showed no inhibitory effect on the actions of adenylosuccinate synthetase and adenylosuccinate lyase. 3. Adenine auxotrophs lacking either adenylosuccinate synthetase or adrenylosuccinate lyase were derived from strain MA406-A-1 and these auxotrophs were found to readily convert formycin B to formycin in replacement culture. From these results, it was estimated that, under the conditions of replacement culture, formycin B would be converted to formycin in vivo by the action of novel enzyme(s) rather than by the action of enzyme system including adenylosuccinate synthetase and adenylosuccinate lyase.

Adenine

Biosynthesis of formycin. Incorporation and distribution of labeled compounds into formycin.

1. Two carbons, carbon-2 and one of carbons-3 to -5, of lysine seemed likely to be incorporated into one of carbon of the chromophore moiety of formycin. 2. From the results of radioisotopic studies with glutamate, gamma-amino-n-butyrate or organic acids related to tricarboxylic acid cycle, the important role of glutamate in the biosynthesis of formycin was strongly suggested. 3. The incorporation of nitrogen(s) of lysine into three nitrogens, including two nitrogens of pyrazole ring, of formycin was suggested by mass spectroscopy. 4. Ribose was estimated as a direct precursor moiety of formycin, whereas the biosynthesis of ribose was shown to occur via the pathway other than hexose monophosphate shunt or glucuronate pathway. 5. In replacement culture, the salvage synthesis of formycin from its chromophore moiety was not observed and it was also evident that the chromophore moiety or pyrazofurin (pyrazomycin) inhibited the biosynthesis of formycin.

Adenine