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A method for the partial purification of the pyruvate-uridine-diphospho-N-acetylglucosamine transferase from Staphylococcus epidermidis is presented. Some properties of the enzyme are discussed.

Wishnow, Rodney M. (Washington University School of Medicine, St. Louis, Mo.), Jack L. Strominger, Claire H. Birge, and Robert H. Threnn. Biochemical effects of novobiocin on Staphylococcus aureus. J. Bacteriol. 89:1117-1123. 1965.-Novobiocin induced accumulation in Staphylococcus aureus of uridine nucleotide precursors of the cell-wall glycopeptide, of a cytidine nucleotide precursor of the teichoic acid in the cell wall, and of a large number of other ultraviolet-absorbing materials. These substances accumulated promptly on addition of the antibiotic at levels near the growth-inhibitory concentration. Isotopic experiments showed that, in addition to inhibiting cell-wall synthesis, novobiocin also inhibited both protein and nucleic acid synthesis. The lack of selectivity in the action of novobiocin is similar to that previously observed with gentian violet, and is quite different from the selective inhibition of cell-wall synthesis observed with penicillin or with d-cycloserine. Although inhibition of cell-wall synthesis is not excluded as one of the primary lethal effects of novobiocin, it is also possible that all of the observed phenomena are secondary to some other primary metabolic lesion.

The staphylolytic enzyme recently isolated from cultures of a Chalaropsis species by Hash is shown to be an acetylmurainidase that cleaves all the glycosidic linkages of N-acetylmuramic acid and N,O-diacetylmuramic acid in the cell wall of Staphylococcus aureus strain Copenhagen. It is similar in specificity to the "32 enzyme" from Streptomyces albus but it differs from egg-white lysozyme whose activity is inhibited by the presence of O-acetyl groups.

Strominger, Jack L. (Washington University School of Medicine, St. Louis, Mo.), and Claire H. Birge. Nucleotide accumulation induced in Staphylococcus aureus by glycine. J. Bacteriol. 89:1124-1127. 1965.-High concentrations of glycine induce accumulation of four uridine nucleotides in Staphylococcus aureus. Investigations of their structure suggest that these compounds are uridine diphosphate (UDP)-acetylmuramic acid, UDP-acetylmuramyl-gly-d-glu-l-lys, UDP-acetylmuramyl-l-ala-d-glu-l-lys and UDP-acetylmuramyl-gly-d-glu-l-lys-d-ala-d-ala. The mechanism by which glycine may induce uridine nucleotide accumulation and protoplast formation is discussed.

Antisera, prepared against formalin-killed cells of Staphylococcus aureus, strain Copenhagen, agglutinated the cell walls of this strain. The agglutination was inhibited by the teichoic acid from the cell wall of this strain, by any degradation product of this teichoic acid which contained the alpha-acetylglucosaminyl-ribitol unit, by alpha-phenyl-acetylglucosaminide, and by N-acetylglucosamine, but not by a large number of other haptens related to the cell wall. In quantitative experiments, however, only 40 to 50 per cent of antibody adsorption to cell wall could be inhibited by teichoic acid or by N-acetylglucosamine. The alpha-acetylglucosaminyl-ribitol unit in the teichoic acid is, therefore, an important immunological determinant in the cell wall of this strain, although other immunological specificities may also exist. The cell walls were also agglutinated by heterologous antisera prepared against streptococcal Group A carbohydrate or against horse serum azophenyl-beta-acetylglucosaminide. The heterologous agglutination, however, was specific for the beta-acetylglucosaminyl-ribitol units in the teichoic acid.

C(55)-isoprenoid alcohol phosphokinase, a butanol-soluble enzyme from the membrane of Staphylococcus aureus, has been purified to homogeneity by the inclusion of organic solvents in all of the techniques used. By polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, the enzyme is a single polypeptide chain of molecular weight 17,000. The amino-acid analysis reveals an unusually high content of nonpolar amino acids (58%), the largest amount so far reported in any protein. When equilibrated in butanol-water, enzyme apoprotein is found in the butanol layer. The influence of sodium chloride indicates that the lipid requirement of the enzyme is nonelectrostatic in nature.

A method for the partial purification of the uridine diphospho-N-acetylglucosamine-enolpyruvate reductase from Staphylococcus epidermidis is presented. Some properties of the enzyme, including its dependence on monovalent cation and flavine adenine dinucleotide, are discussed.