Phospholipids of Streptococcus faecalis

Autoradiograms of total lipid extracts from Streptococcus faecalis ATCC 9790, harvested in the stationary phase from a medium containing (32)P-orthophosphate, showed six major spots. The corresponding compounds were identified as diphosphatidylglycerol (possibly with a penta acyl structure); phosphatidylglycerol; a provisionally identified mixture of alanylphosphatidylglycerol and of the 2'-lysyl-derivative of phosphatidylglycerol; the 3'-lysyl-derivative of phosphatidylglycerol, probably together with some arginylphosphatidylglycerol; a diglucosyl derivative of phosphatidylglycerol; and a compound which was tentatively identified as the 2',3'-dilysyl derivative of phosphatidylglycerol.     

Folate metabolism in Streptococcus faecalis

The possibility that the inability of Streptococcus faecalis to utilize 5-methyltetrahydropteroylglutamate or pteroyltriglutamate might be due to permeability was investigated. Whereas the former was taken up by S. faecalis cells growing on pteroylglutamic acid, the latter was not. No subsequent conversion of the 5-methyltetrahydropteroylglutamate took place and accumulation, which was against a considerable concentration gradient, was inhibited by fluoride. It would thus appear to be an active process.     

D-alanine carboxypeptidase from Streptococcus faecalis

A particulate D-alanine carboxypeptidase that can cleave the terminal residue of D-alanine from UDPMurNAc-L-ala-D-isoglu-L-lys-D-ala-D-ala was isolated from $\text{Streptococcus faecalis}$. The enzyme was inhibited by penicillin G non-competitively with a K_i of 0.8 μM.The carboxypeptidase was solubilized with Triton X-100 without loss of catalytic activity. In this form it could also be inhibited by penicillin G.     

Molar Growth Yield of Streptococcus faecalis on Pyruvate

Y(pyruvate) was 17.3, similar to Y(arginine), for Streptococcus faecalis 6783 grown statically in a complex medium in 1 atm of air.     

Sodium-stimulated ATPase in Streptococcus faecalis.

We measured Na+-stimulated ATPase activity in a mutant of Streptococcus faecalis defective in the generation of proton motive force. The activity in membrane vesicles was 62.1 +/- 5.9 nmol of phosphate produced per min per mg of protein when cells were grown on medium containing 0.12 M Na+. Activity decreased as the concentration of Na+ in the growth medium decreased. The decrease in enzyme activity corresponded to the decrease in transport activity for Na+ in both whole cells and membrane vesicles. The effects of pH on both activities were identical. Thus, it is suggested that Na+ movement is mediated by this enzyme. Sodium extrusion and ATPase activity in the wild-type strain were markedly lower than those observed in the mutant strain. Elevated activities of both Na+ extrusion and Na+-stimulated ATPase could be detected in the wild-type strain when cells were grown in the absence of proton motive force. Thus, we propose that the level of ATPase is increased by dissipation of the proton motive force.     

Recombination-deficient mutant of Streptococcus faecalis.

An ultraviolet radiation-sensitive derivative of Streptococcus faecalis strain JH2-2 was isolated and found to be deficient in recombination, using a plasmid-plasmid recombination system. The strain was sensitive to chemical agents which interact with deoxyribonucleic acid and also underwent deoxyribonucleic acid degradation after ultraviolet irradiation. Thus, the mutant has properties similar to those of recA strains of Escherichia coli.     

Fumarate reductase activity of Streptococcus faecalis

Some characteristics of a fumarate reductase from Streptococcus faecalis are described. The enzyme had a pH optimum of 7.4; optimal activity was observed when the ionic strength of the phosphate buffer was adjusted to 0.088. The K(m) value of the enzyme for reduced flavin mononucleotide was 2 x 10(-4)m as determined with a 26-fold preparation. In addition to fumarate, the enzyme reduced maleate and mesaconate. No succinate dehydrogenase activity was detected, but succinate did act as an inhibitor of the fumarate reductase activity. Other inhibitors were malonate, citraconate, and trans-, trans-muconate. Metal-chelating agents did not inhibit the enzyme. A limited inhibition by sulfhydryl-binding agents was observed, and the preparations were sensitive to air oxidation and storage. Glycine, alanine, histidine, and possibly lysine stimulated fumarate reductase activity in the cell-free extracts. However, growth in media supplemented with glycine did not enhance fumarate reductase activity. The enzymatic activity appears to be constitutive.