PROCEDE D'OBTENIR L'ANTIBIOTIQUE A-287

1455613 Antibiotic A-287 ELI LILLY & CO 11 Jan 1974 [22 Jan 1973] 01502/74 Heading C2A A process for producing antibiotic A-287 comprising factor A and/or factor B comprises cultivating Actinoplanes utahensis NRRL 5614 in a culture medium containing assimilable sources of carbohydrate, nitrogen and inorganic salts under submerged aerobic fermentation conditions until a substantial amount of antibiotic activity is produced, and separating the A-287 antibiotic mixture from the culture medium. The factors are separated from the mixture and isolated from each other by wellknown techniques, e.g. counter-current distribution and thin-layer chromatography. The antibiotic A-287 factor A: (a) is a white amorphous compound, (b) does not melt below about 220‹ C., (c) has an elemental composition of 44À98% carbon, 6À15% hydrogen, 10À28% nitrogen, 26À37% oxygen and 4À15% ash, (d) has an apparent molecular weight in the range of 1500 (1475À73), as determined in methanol, (e) in mineral oil mull has an I.R. spectrum as indicated in Fig. 1 (not shown), (f) in water absorbs in the U.V. region with maxima as follows: # max. 220 mÁ (# 9500; shoulder), # max. 273 mÁ (# 2700), (g) is soluble in lower alcohols and water, (h) is insoluble in many organic solvents, (i) has an approximate ratio of amino acids as follows: serine (1), glutamic acid (1), glycine (2), alanine (1), valine (2), cysteine (1), isoleucine (1), leucine (2), tryptophan (1) and unknown (2), (j) has the following Rf values in the paper chromatography systems indicated below, using Bacillus subtilis ATCC 6633, Sarciena lutea ATCC 9341 or Staphylococcus aureus ATCC 6538 as detection organisms: Rf value Solvent system 0À17 Butanol saturated with water. 0À70 Butanol saturated with water plus 2% p-toluenesulphonic acid (p- TSA). 0À39 Butanol saturated with water plus 2% p-TSA and 2% piperidine. 0À72 80% ethanol with 1À5% NaCl. Paper is impregnated with soln. of 0À95M Na 2 SO 4 and 0À05M NaHSO 4 H 2 O. 0À68 Methanol : 0À1M HCl (3 : 1). The antibiotic A-287 factor B: (a) is a white amorphous compound, (b) chars at about 200‹ C., (c) has an approximate elemental composition of 49À96% carbon, 6À98% hydrogen, 9À90% nitrogen, 23À86% oxygen and 4À66% ash, (d) has an apparent molecular weight in the range 2800 (2781À28) as determined in methanol, (e) in mineral oil mull has an I.R. spectrum as indicated in Fig. 2 (not shown), (f) in water absorbs in the U.V. with maxima as follows: # max. 220 mÁ (# 9200; shoulder), # max.