Evidence for the de novo synthesis of the alpha-amylase of Pseudomonas saccharophila.
Eisenstadt, Jerome M. (Brandeis University, Waltham, Mass.) and Harold P. Klein. Evidence for the de novo synthesis of the alpha-amylase of Pseudomonas saccharophila. J. Bacteriol. 82:798-807. 1961.-Chloramphenicol at a concentration of 20 mug per ml inhibited the appearance of the inducible alpha-amylase of Pseudomonas saccharophila. This inhibition was observed when induction was attempted in buffer or in a complete medium. Preinduced cells were also prevented from forming this enzyme under similar conditions. Under all the conditions tested, there was no lag in chloramphenicol inhibition, thus suggesting an absence of any protein precursor in amylase formation. Cells suspended in a complete medium without a nitrogen source lost their capacity to form this enzyme when subsequently induced in buffer. When cells were grown in the presence of radioactive sulfate and then subjected to starvation, the radioactivity of the amino acid pool diminished only slightly. However, examination of the free amino acid pool by paper chromatography showed that the loss of enzyme inducibility was accompanied by the disappearance of glutamine, aspartic acid, and a third, unidentified, compound. Enzyme-forming ability was restored by the addition, to starved cells of casein hydrolysate, glutamate, glutamine, or aspartate. Other amino acids tested were ineffective in this regard. When cells were induced in buffer in the presence of labeled methionine, amylase was formed at a linear rate over a 3-hr period. Furthermore, both the cellular proteins and the extracellular amylase became labeled at a linear rate. These observations are discussed in relation to the problem of protein turnover, and are interpreted as evidence for the de novo synthesis of alpha-amylase in this organism.
EISENSTADT
J M
JM
KLEIN
H P
HP
D000596:Amino Acids; D000681:Amylases / Q000378:metabolism*; D001224:Aspartic Acid; D042621:Comamonadaceae; D008970:Molecular Weight; D009584:Nitrogen; D011506:Proteins; D011549:Pseudomonas / Q000378:metabolism*; D000516:alpha-Amylases
D000596:Amino Acids; D011506:Proteins; D001224:Aspartic Acid; D000681:Amylases; D000516:alpha-Amylases; D009584:Nitrogen
AMYLASES/metabolism; PSEUDOMONAS/metabolism