THE DEVELOPMENT OF RESISTANCE TO TERRAMYCIN BY BACT. LACTIS AEROGENES (AEROBACTER AEROGENES).
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Aerobacter aerogenes strain PRL-R3 possesses inducible enzyme pathways for the catabolism of d-xylose and d-arabitol. d-Xylose is the apparent inducer for d-xylose isomerase and d-xylulokinase. d-Arabitol is the apparent inducer for d-arabitol dehydrogenase and a separate d-xylulokinase. Both kinases had similar K(m) values and substrate specificities, and could not be separated by sucrose gradient centrifugation or polyacrylamide gel electrophoresis. They could be differentiated, however, by their separate regulation, their inhibition by antisera, and by the cold sensitivity of the kinase of the d-arabitol catabolic pathway.
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Mutants of Aerobacter aerogenes devoid of acetate kinase and phosphotransacetylase activities were isolated by selection for resistance to fluoroacetate on lactate medium. The mutants were used to study the role of the acetate kinase-phosphotransacetylase system in growth on acetate and glucose. Acetate kinase-negative and phosphotransacetylase-negative mutants were unable to grow on acetate minimal medium. Their growth rates on glucose minimal medium were identical with that of the parent strain under aerobic conditions, but lower growth rates were observed in the mutant strains during anaerobic growth on glucose medium. The mutants were unable to incorporate [2-(14)C]-acetate rapidly while growing on glycerol. Variations in acetate kinase and phosphotransacetylase levels during growth on glucose were studied. The specific activities of the enzymes increased approximately fivefold during aerobic growth on glucose in batch culture. The enzyme levels were also studied during anaerobic growth on glucose at constant pH (pH 5.8 and 7.0). Smaller increases in specific activities were found under these conditions. The role of acetate in the induction of the diacetyl (acetoin) reductase was investigated using a mutant deficient in both acetate kinase and phosphotransacetylase. The effect of pH on the induction of this enzyme during growth on glucose under anaerobic conditions was tested. The data support the idea that free acetic acid is the inducer for the enzymes of the butanediol-forming pathway in A. aerogenes.
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The data presented in this paper indicate operation of different mechanisms for acetate oxidation by A. aerogenes, depending on the carbon source used for growth. The mechanism for citrate-grown cells appears to involve a conventional citric acid cycle, whereas acetate-grown cells appear to incorporate acetate carbon more readily via a dicarboxylic acid cycle.
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