The addition of short-chain fatty acids stimulates the production of spiramycin by Streptomyces ambofaciens cultivated on dextrins and ammonium chloride. The fatty acids were activated by two enzymatic systems. The first system (acyl-CoA synthetases) was present only during the exponential phase. The second system (acylkinases coupled with acylphosphotransferases) was synthesized during the growth phase and during the stationary phase, in which spiramycin production started. Short-chain fatty acids induced the synthesis of acylkinases and acylphosphotransferases. Added at the beginning of cultures, they increased the specific activity of these enzymes during the exponential growth phase. Added at the early stationary phase, the specific activity of these enzymes and of the spiramycin production increased. Excess ammonium in the culture considerably lowered the specific activity of acylkinases synthesized in the stationary phase, when spiramycin production started. This ammonium effect can be reduced by the addition of short-chain fatty acids.
Spiramycin biosynthesis in Streptomyces ambofaciens was stimulated in the presence of valine or by sequential addition of some short-chain fatty acids to a culture medium containing an ammonium salt as source of nitrogen. Acetate kinase and acetyl-CoA carboxylase, enzymes that catalysed the formation of precursors of spiramycin biosynthesis (acetyl-CoA and malonyl-CoA), were detected during the active growth and antibiotic production phases. In this latter phase a higher level of acetyl-CoA carboxylase activity was observed with valine (1.02 mumol.min-1.mg protein-1) than with ammonium (0.05 mumol.min-1.mg protein-1) as nitrogen source, while the evolution and the level of acetate kinase activity were the same in both media. Successive addition of acetate and isobutyrate stimulated highly and weakly the acetyl-CoA carboxylase and acetate kinase activity, respectively.
Growth of Streptomyces ambofaciens in the presence of valine led to a greater production of spiramycin and excretion of short-chain fatty acids compared with growth on ammonium ions as nitrogen source. We determined the activities of enzymes that lead to the formation of malonyl-CoA, a key intermediate in the biosynthesis of spiramycin. In valine-grown cultures, the activities of acetyl-CoA carboxylase and pyruvate dehydrogenase were increased during the antibiotic production phase. During this idiOphaSe, oxaloacetate dehydrogenase activity was much higher in cultures growing on ammonium compared with valine. Two pathways are proposed concerning the mode of malonyl-CoA formation : a route via oxaloacetate dehydrogenase following growth on ammonium, and a route via acetyl-CoA carboxylase after growth on valine.
The production of cephamycin C by Streptomyces cattleya varies with the use of asparagine, glutamine or ammonium as nitrogen sources. Hydroxylase and expandase activities were demonstrated for the first time with this species. A study of the biosynthetic regulation of these enzymes by two different nitrogen sources, glutamine and asparagine, was carried out. Asparagine proved to be a better nitrogen source, both for enzymatic biosynthesis and production of cephamycin C. Moreover, an excess of asparagine in the culture environment provokes, simultaneously, a reduction in cephamycin C production and a decrease in the biosynthesis of expandase and hydroxylase.
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