L-Asparaginase of Klebsiella aerogenes. Activation of its synthesis by glutamine synthetase.
An L-asparaginase has been purified some 250-fold from extracts of Klebsiella aerogenes to near homogeneity. The enzyme has a molecular weight of 141,000 as measured by gel filtration and appears to consist of four subunits of molecular weight 37,000. The enzyme has high affinity for L-asparagine, with a Km below 10(-5) M, and hydrolyzes glutamine at a 20-fold lower rate, with a Km of 10(-3) M. Interestingly, the enzyme exhibits marked gamma-glutamyltransferase activity but comparatively little beta-aspartyl-transferase activity. A mutant strain lacking this asparaginase has been isolated and grows at 1/2 to 1/3 the rate of the parent strain when asparagine is provided in the medium as the sole source of nitrogen. This strain grows as well as the wild type when the medium is supplemented with histidine or ammonia. Glutamine synthetase activates the formation of L-asparaginase. Mutants lacking glutamine synthetase fail to produce the asparaginase, and mutants with a high constitutive level of glutamine synthetase also contain the asparaginase at a high level. Thus, the formation of asparaginase is regulated in parallel with that of other enzymes capable of supplying the cell with ammonia or glutamate, such as histidase and proline oxidase. Formation of the asparaginase does not require induction by asparaginase and is not subject to catabolite repression.
D000469:Alkaline Phosphatase / Q000378:metabolism; D001215:Asparaginase / Q000096:biosynthesis* / Q000302:isolation & purification; D002455:Cell Division; D004789:Enzyme Activation / Q000187:drug effects; D005696:Galactosidases / Q000378:metabolism; D005838:Genotype; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism* / Q000494:pharmacology; D006638:Histidine Ammonia-Lyase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007709:Klebsiella / Q000201:enzymology*; D009113:Muramidase; D009154:Mutation; D010641:Phenotype; D013045:Species Specificity
D000469:Alkaline Phosphatase; D005696:Galactosidases; D009113:Muramidase; D001215:Asparaginase; D006638:Histidine Ammonia-Lyase; D005974:Glutamate-Ammonia Ligase