Effect of Alanine, Leucine, and Fructose on Lysyl-Transfer Ribonucleic Acid Ligase Activity in a Mutant of Escherichia coli K-12

Abstract

A mutant of Escherichia coli K-12 was examined which has growth medium-dependent lysyl-transfer ribonucleic acid (tRNA) ligase activity. In minimal medium or 0.5% yeast extract, the activity of the enzyme in the mutant strain was 5 to 10% of wild type. However, when the mutant was grown in a highly enriched medium, such as AC broth (Difco), the activity of the mutant ligase increased 10- to 20-fold. We found that the supplementation of 0.5% yeast extract by l -alanine plus d -fructose replaces the need for the highly enriched medium. Fructose plus l -leucine and fructose plus l -α-amino- n -butyric acid were also stimulatory, but not as effective as fructose and alanine. With minimal medium, a combination of carbohydrate (fructose or glucose) plus alanine and leucine was required to replace the enriched medium. The most effective combination was fructose, glucose, alanine, and leucine. Lysyl-tRNA ligase was stimulated 1.5 to 2-fold in the wild-type strain or Hfr H (Hayes) by fructose plus alanine when these strains were cultured in 0.5% yeast extract. Experiments employing the combined technique of density labeling with D 2 O and isopycnic centrifugation in cesium chloride indicated that the increased activity of lysyl-tRNA ligase observed in AC broth or in the presence of fructose, glucose, alanine, and leucine is due to the synthesis of new enzyme.