Lividomycins 5-phosphate

The structure, lividomycin 5"-phosphate, was furthermore proved by chemical synthesis. Penta-N-(benzyloxycarbonyl) lividomycin A was prepared from lividomycin A by the usual Schotten-Baumann procedure in a yield of 95% it had m.p. 135-150° (dec.). Acetonation with 2,2-dimethoxypropane in N,N-dimethylformamide in the presence of p-toluenesulfonic acid at 110° for 4 hours afforded the tri-O-isopropylidene derivative of N-(benzyloxycarbonyl)lividomycin A in 49% yield m.p. 129-133° (dec.). Phosphorylation of the sole primary hydroxyl group in the D-ribose moiety of -(benzyloxycarbonyl) lividomycin A with diphenyl phosphorochloridate in dry pyridine gave penta-N-(benzyloxy-... 

Phosphotransferase I phosphorylates not only the 3 -hydroxyl group, but also the 5"-hydroxyl function of lividomycin and 3, 4 -dideoxyribostamycin. The exclusive attack at the 3 -position of ribostamycin indicates that this position is steri-cally better located for the phosphate-transferring center of the enzyme than the 5"-hydroxyl group. A 5"-0-phosphotransferase that reacts also in this position of ribostamycin has been isolated f tom Pseudomonas. The interesting point of these S"-0-phosphorylations is that the 1-amino-HABA moiety in butirosin obviousiy... 

Escherichia coli K12 ML1629, . coU K12 ML1410 R81, and E. coli K12 J5 Rll-2 showed similar resistant patterns to aminoglycosidic antibiotics, and the S-100 fraction contained kanamycin-neomycin phosphate transferase I. This enzyme solution also inactivated lividomycins A and B (6 and 7) in the presence of adenosine 5 -triphosphate. A crude powder of the inactivated lividomydn A extracted by column chroma-... 

Partial purification of the lividomycin-inactivating enzyme was attempted by Mitsuhashi and coworkers, who briefly described their discovery that the enzyme obtained by fractionation with ammonium sulfate and column chromatography on Sephadex G-lOO inactivates lividomycins A and B, but not kanamycin A, indicating involvement of two different enzymes in the phosphorylation of lividomycin and kanamycin. However, it was definitely proved by H. Umezawa and coworkers that kanamycin-neomycin phosphate transferase I phosphorylates the 5"-hydroxyl group of lividomycins. The observation by Mitsuhashi and coworkers was probably occasioned by instability of the enzyme and the higher activity of the enzyme in phosphorylating lividomycins than in phosphorylating kanamycin A. 

Kanamycin-neomycin phosphate transferase I was successfully purified by affinity chromatography with lividomycin A-Sepharose 4B. Livi-... 

The involvement of kanamycin-neomycin phosphate transferase II in the mechanism of resistance was demonstrated by the fact that Escherichia coli KI2 JR66/W677 is inhibited by 3, 4 -dideoxybutirosin B, 3, 4 -dideoxyribostamycin,- 3, 4 -dideoxyneamine, and lividomycins that do not undergo this enzyme reaction. 

Cross-resistance between kanamycins and lividomycins is frequently observed in staphylococci, suggesting the presence of kanamycin-neomycin phosphate transferase I. However, phosphorylation of butiro-sins or lividomycins by a staphylococcus enzyme has not yet been studied. 

Lividomycins lack a 3 -hydroxyl group that could undergo the reaction of a kanamycin-neomycin phosphate transferase, but the 5"-hydroxyl group is phosphorylated by kanamycin-neomycin phosphate transferase... 

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