Lividomycin Resistance in Staphylococci by Enzymatic Phosphorylation

Abstract

Enzymatic inactivation of lividomycin (LV) was attempted with nine clinical isolates of staphylococci including LV-susceptible and -resistant strains. LV inactivation and the incorporation into LV of 32P from y-32P-adenosine triphosphate were demonstrated in the presence of cell-free extracts from LV-resistant strains but not from LV-susceptible ones. The enzyme was purified approximately 82-fold from a resistant Staphylococcus aureus strain by means of ammonium sulfate fractionation and column chromatography. Some properties of the partially purified LV-phosphorylating enzyme were quite similar to those of an enzyme from Escherichia coli carrying an R factor conferring LV resistance, and the phosphorylated product of the drug was also found to be identical with that produced by E. coli carrying an R factor, i.e., 5"-phosphoryl-LV.Many enzymes which inactivate aminoglycosidic antibiotics have been reported in drugresistant strains of bacteria of several species (1, 12). We reported that the lividomycin (LV)-inactivating enzyme capable of phosphorylating the drug was demonstrated in resistant strains of Pseudomonas aeruginosa (9) and of Escherichia coli carrying an R factor (19). Further studies indicated that the inactivated LV was a monophosphorylated product, in which the Dribose moiety of LV was phosphorylated (10,20).In a survey of Staphylococcus aureus, LVresistant strains whose growth was not inhibited by 50 qig of LV/ml were isolated at a frequency of 23.4% (6). This paper deals with some properties of the LV-inactivating enzyme and the relation between LV resistance and the enzyme activity.MATERIALS AND METHODS Bacterial strains. The nine clinical isolates used included three LV-resistant and three LV-susceptible strains of S. aureus and three LV-resistant strains of S. epidermidis. These strains were selected at random from our stocks.Preparation of the S-105 fraction. Glucose-peptone broth (pH 7.2) consisted of 10 g of peptone, 5 g of NaCl, 3 g of glucose, 4 g of yeast extract, and 1,000 ml of water. Each strain was cultured in 2 liters of glucose-peptone broth at 35 C for 5 h with shaking, after 18 h of precultivation in the same broth at 37 C without shaking.

New Plasmid-Mediated Nucleotidylation of Aminoglycoside Antibiotics in Staphylococcus aureus

“…This phenomenon was later demonstrated to be plasmid mediated (16). A lividomycin phosphotransferase has also been isolated from S. aureus strains (7). The phosphorylated product of this drug was found to be identical to that ofEscherichia coli carrying an R plasmid that confers lividomycin resistance (8).…”

Phosphorylation of Kanamycin, Lividomycin A, and Butirosin B by Providencia stuartii

“…Resistance transfer factor enzyme phosphorylation, involving several phosphotransferases, has been responsible for resistance to high concentrations of aminoglycoside antibiotics in E. coli, Pseudomonas aeruginosa, and Staphylococcus aureus (3,4,7,8,20). Neomycin-kanamycin phosphotransferase I has been shown to phosphorylate the 3'-hydroxyl group of the aminohexose moiety of kanamycin and neomycin (5,15,16) and the 5"-hydroxyl group of the ribose moiety of lividomycin A (17).…”