Lactate dehydrogenase activity in certain strains of Staphylococcus aureus.
Lactate dehydrogenase (LDH) was studied in phage-propagating strains 29, 3A, 6, 81, and 42D of Staphylococcus aureus selected from the five groups in the International-Blair series. Cells were cultivated in Brain Heart Infusion (Difco) under nearly anaerobic conditions and were harvested near the end of the log phase. LDH activity was maximal at the end of the exponential growth period and was measured spectrophotometrically by reduction of p-nitro-blue tetrazolium, with phenazine methosulfate as a coupling agent. Crude enzyme extracts were prepared both by an acetone extraction technique and by sonic treatment. LDH activity for these enzyme preparations was determined by the colorimetric method mentioned and also by measuring the rate of nicotinamide adenine dinucleotide reduction at 340 mmu. The order of activity observed, by use of both assay methods, was 29 > 81 > 6 > 3A > 42D. LDH forms (possibly isoenzymes) for each of 15 strains, which represent the five phage-propagating groups of the International-Blair series, were separated by acrylamide gel electrophoresis. Five forms were distinguished and arbitrarily numbered on the basis of their rate of migration, no. 5 being the slowest component. No one strain had more than four, nor fewer than two, LDH forms. Form 3 appeared in 13 of the 15 strains and was followed in frequency by no. 2, 1, 4, and 5.
D003470:Culture Media; D003720:Densitometry; D004589:Electrophoresis, Disc; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D013053:Spectrophotometry; D013210:Staphylococcus / Q000201:enzymology*
D003470:Culture Media; D007527:Isoenzymes; D009243:NAD; D007770:L-Lactate Dehydrogenase