Relationship between nitrogen assimilation and cephalosporin synthesis inStreptomyces clavuligerus |
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Authors: | Alfredo F. Brana Saul Wolfe Arnold L. Demain |
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Affiliation: | (1) Fermentation Microbiology Laboratory, Department of Applied Biological Sciences, Massachusetts Institute of Technology, 02139 Cambridge, MA, USA;(2) Department of Chemistry, Queens University, K7L 3N6 Kingston, Ontario, Canada;(3) Present address: Departamento de Microbiología, Universidad de Oviedo, Oviedo, Spain |
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Abstract: | The levels of three enzymes of the -lactam antibiotic pathway and overall cephalosporin production were subject to nitrogen source repression inStreptomyces clavuligerus. The specific activities of isopenicillin N synthetase (cyclase) and deacetoxycephalosporin C synthetase (expandase) measured during the exponential phase depended on the nitrogen source employed, following a pattern that roughly correlated with the corresponding antibiotic production. The effects on isopenicillin N epimerase (epimerase) activities were less marked than those on the cyclase and expandase.Production of cephalosporins and enzymatic activities were not related to the growth rate of the cultures. Glutamate, glutamine and alanine inhibited production when added to resting cell systems, while lysine and -aminoadipate were stimulatory. No clear relationship could be drawn between cephalosporin production or -lactam synthetase activities and the activities of enzymes of ammonium assimilation (glutamine synthetase, glutamate synthase and alanine dehydrogenase).The intracellular pools of free glutamine, alanine and ammonium were the only ones markedly affected by the nitrogen source in the wild type and mutants, but these amino acids did not seem to play an obvious role as intracellular mediators of nitrogen control.Abbreviations DCW dry cell weight - GS glutamine synthetase - GOGAT glutamate synthase - ADH alanine dehydrogenase - HPLC high performance liquid chromatography |
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Keywords: | Nitrogen assimilation Cephalosporin Nitrogen regulation Ammonium repression Streptomyces clavuligerus |
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