Chorismate mutase from Streptomyces aureofaciens: a heat-stable enzyme.
Chorismate mutase from Streptomyces aureofaciens was purified 12-fold. This enzyme preparation did not show any activity when tested for anthranilate synthetase, prephenate dehydrogenase, or prephenate dehydratase. The catalytic activity of chorismate mutase has a broad optimum between pH 7 and 8. The initial velocity data followed regular Michaelis-Menten kinetics with a K(m) of 5.3 x 10(-4) M, and the molecular weight of the enzyme was determined by sucrose gradient centrifugation to be 50,000. Heat inactivation of chorismate mutase, which occurs above temperatures of 60 C, is reversible. The enzyme activity can be restored even when chorismate mutase is treated at the temperature of a boiling-water bath for 15 min. Heat-denatured and renatured enzymes showed the same Michaelis constant and the same molecular weight as the native enzyme. l-Phenylalanine, l-tyrosine, l-tryptophan, and metabolites of the aromatic amino acid pathway were tested as potential modifiers of chorismate mutase activity. The activity of the enzyme was inhibited by none of these substances. Chorismate mutase of S. aureofaciens was not repressed in cells grown in minimal medium supplemented with l-phenylalanine, l-tyrosine, or l-tryptophan.
D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D003509:Cyclohexanecarboxylic Acids; D003510:Cyclohexanes; D005246:Feedback; D006358:Hot Temperature; D006836:Hydro-Lyases / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007535:Isomerases / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000302:isolation & purification* / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D010649:Phenylalanine / Q000494:pharmacology; D011489:Protein Denaturation; D011773:Pyruvates; D013237:Stereoisomerism; D013302:Streptomyces / Q000201:enzymology*; D013304:Streptomyces aureofaciens / Q000201:enzymology / Q000254:growth & development; D000637:Transaminases / Q000378:metabolism; D014364:Tryptophan / Q000494:pharmacology; D014443:Tyrosine / Q000494:pharmacology; D014753:Vinyl Compounds; D062367:ortho-Aminobenzoates
D003509:Cyclohexanecarboxylic Acids; D003510:Cyclohexanes; D009097:Multienzyme Complexes; D011773:Pyruvates; D014753:Vinyl Compounds; D062367:ortho-Aminobenzoates; D014443:Tyrosine; D010649:Phenylalanine; D014364:Tryptophan; D010088:Oxidoreductases; D000637:Transaminases; D006836:Hydro-Lyases; D007535:Isomerases