Regulation of glutamine synthetase. V. Partial purification and properties of glutamine synthetase from Bacillus licheniformis.
The glutamine synthetase of Bacillus licheniformis has been obtained at about 15% purity. Sucrose gradient centrifugation gave a molecular weight value of approximately 612,000. Both l- and d-glutamate can be utilized as substrates in the biosynthetic reaction, although the l isomer was five times more active. The requirement for adenosine triphosphate (ATP) can be partially replaced by guanosine or inosine triphosphates, but not by cytidine or uridine triphosphates. The Mn(++) was required for activity, and the requirement cannot be satisfied with Mg(++). Maximal activity of the biosynthetic reaction was observed when ATP and Mn(++) were present in equimolar amounts. An excess of either reactant gave less activity. However, other purine and pyrimidine nucleotides, when added in combination with ATP, can partially substitute for ATP in attaining the equimolar ratio of nucleotide to Mn(++). A complex of ATP and Mn(++) is the preferred form of substrate. The B. licheniformis enzyme catalyzes the glutamyl transfer reaction but at a much slower rate than the Escherichia coli glutamine synthetase. Either adenosine diphosphate (ADP) or ATP can activate the glutamotransferase, although ADP is more active.
D000217:Acyltransferases / Q000378:metabolism; D000227:Adenine Nucleotides / Q000494:pharmacology; D000643:Ammonium Chloride / Q000494:pharmacology; D001224:Aspartic Acid / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D003597:Cytosine Nucleotides / Q000494:pharmacology; D004586:Electrophoresis; D005971:Glutamates / Q000378:metabolism / Q000494:pharmacology; D005973:Glutamine; D006150:Guanine Nucleotides / Q000494:pharmacology; D006877:Hydroxamic Acids / Q000096:biosynthesis; D008025:Ligases / Q000032:analysis / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D009711:Nucleotides / Q000494:pharmacology; D011770:Pyruvate Kinase / Q000494:pharmacology; D011773:Pyruvates / Q000494:pharmacology; D014500:Uracil Nucleotides / Q000494:pharmacology
D000227:Adenine Nucleotides; D003597:Cytosine Nucleotides; D005971:Glutamates; D006150:Guanine Nucleotides; D006877:Hydroxamic Acids; D009711:Nucleotides; D011773:Pyruvates; D014500:Uracil Nucleotides; D000643:Ammonium Chloride; D005973:Glutamine; D001224:Aspartic Acid; D008345:Manganese; D000217:Acyltransferases; D011770:Pyruvate Kinase; D008025:Ligases; D008274:Magnesium