| 摘要 |
<p>1470407 Antibiotics GKUPPO LEPETIT SpA 25 July 1975 [27 July 1974] 33283/74 Heading C2A Antibiotic substances known as metabolite A metabolite B (referred to herein as gardimycin) and metabolite C are prepared by activating either Actinoplanes garbadinensis ATCC 31049, or Actinoplanes ligurial ATCC 31048 and mutants or equivalents thereof under aerobic conditions in an aqueous medium containing assimilable sources of carbon, nitrogen and inorganic salts until substantial antibiotic activity is present and recovering the antibiotics from the medium. The metabolites may be purified and isolated by known techniques. Gardimycin as the monosodium salt has the following characteristics :- 1. M.Pt : 260‹ C. (decomposition) 2. M.Wt : 2005-2168 3. Elemental analysis 4. U.V. absorption bands Solvent 2 max.(m) E 1% 1 cm methanol 273 (shoulder) 280 26 299 24 sodium hydroxide 0À1N 273 (shoulder) 279 26 288 22 hydrochloric acid 0À1N 273 (shoulder) 279 26 288 22 phosphate buffer 273 (shoulder) pH 7À38 279 24 288 20 The complete spectrum is illustrated in Fig. 1 (not shown). 5. I.R. spectrum; characteristic absorption bands in mijol at the following frequencies (cm.<SP>-1</SP>) : 3280, 2920-2840 (mijol), 1650, 1520, 1455 (mijol), 1375 (mijol), 1260, 1045, 999, 720. The complete spectrum is illustrated in Fig. 2 (not shown). 6. Specific rotation 7. Solubility Soluble in water, aqueous sodium bicarbonate, diluted aqueous solutions of alkali hydroxides, hot methanol, dimethylformamide, dimethylsulfoxide, glacial acetic acid. Insoluble in dilute mineral acids, benzene, acetone, chloroform, carbon tetrachloride, (C 2-6 ) aliphatic alkanols and tetrahydrofuran. 8. Characteristic reactions 9. Ionizable functionsan ionizable function is potentiometrically detected with a pKa = 7À1 (water) and a pKa = (methyl cellosolve : water = 16 : 4). 10. Iso electric point 4À2 (determined by electrofocusing). 11. Rf values chromatography on Whatman paper No. 1; visualization of the spots by microbiological development on Staphylococcus aureus. Eluting System Rf values (1) Butanol saturated with phos- 0À15 phate buffer M/15 pH 6À0 (2) Butanol saturated with water 0À80 containing 2% p-toluenesulfonic acid (3) Butanol saturated with water 0À10 containing 2% of ammonium hydroxide (4) Phosphate buffer M/15 pH 6À0 0À80 saturated with butanol (5) 20% aqueous solution of 0À80 sodium hydroxide (6) Butanol : methanol : water = 0À65 40 : 10 : 20 with 0À75% of methyl orange (7) Butanol : methanol : water = 0À60 40 : 10 : 20 (8) Acetone : water=1 : 1 0À80 (9) Ethyl acetate saturated with 0À00 water 12. t.l.c. on silica gel; visualization of the spots by sulfuric acid and vanilline and microbiological development of Staphylococcus aureus. Eluting system Rf value Ammonium hydroxide : 0À70 ethanol : water = 1 : 8 : 1 13. The antibiotic is further characterized by the following acids which are obtained by hydrolysis with HCl:- Acid Approx. ratio Valine 2 Serine 1 Glycine 2 Glutamic acid 1 Isoleucine 2 Leucine 1 Alanine 1 Tryptophan 1</p> |
