Purification, properties, and regulation of glutamic dehydrogenase of Bacillus licheniformis.
Cell-free extracts of Bacillus licheniformis and B. cereus were found to contain high specific activities of nicotinamide adenine dinucleotide phosphate (NADP)-dependent-l-glutamate dehydrogenase [EC 1.4.1.4; l-glutamate: NADP oxidoreductase (deaminating)]. Maximum specific activities were found in extracts of cells during the late exponential phase of growth when ammonium ion served as the sole source of nitrogen. Extremely low specific activities were detected throughout the growth cycle when l-glutamate or Casamino Acids served as the source of carbon and nitrogen. The enzyme was purified 55-fold from crude extracts of B. licheniformis, and apparent kinetic constants were determined. Sigmoidal saturation kinetics were not observed, and various adenylates had no effect on the enzyme. Repression of enzyme synthesis during growth on l-glutamate or Casamino Acids was partially overcome by additions of glucose or pyruvate, and this apparent derepression was totally abolished by inhibitors of ribonucleic acid and protein synthesis. Similarly, additions of l-glutamate or Casamino Acids to cells growing on glucose-ammonium ion resulted in strong repression of enzyme synthesis. It is suggested that the enzyme serves an anabolic role in metabolism. Nicotinamide adenine dinucleotide-dependent glutamate dehydrogenase activity was not detected in five species of Bacillus, irrespective of nutritional conditions or of the physiological age of cells.
D000596:Amino Acids / Q000378:metabolism; D000643:Ammonium Chloride / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D002474:Cell-Free System; D011232:Chemical Precipitation; D002701:Chloramphenicol / Q000494:pharmacology; D002848:Chromatography, DEAE-Cellulose; D003124:Colorimetry; D003470:Culture Media; D004789:Enzyme Activation; D004794:Enzyme Repression; D005947:Glucose / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism; D008293:Malates / Q000378:metabolism; D008937:Mitomycins / Q000494:pharmacology; D009249:NADP / Q000378:metabolism; D000644:Quaternary Ammonium Compounds; D012293:Rifampin / Q000494:pharmacology; D012997:Solvents; D013045:Species Specificity; D013053:Spectrophotometry; D013431:Sulfates
D000596:Amino Acids; D003470:Culture Media; D005971:Glutamates; D008293:Malates; D008937:Mitomycins; D000644:Quaternary Ammonium Compounds; D012997:Solvents; D013431:Sulfates; D000643:Ammonium Chloride; D009249:NADP; D002701:Chloramphenicol; D005969:Glutamate Dehydrogenase; D005947:Glucose; D012293:Rifampin