Patent ES-255258-A1 (ES)
Title: Chalcomycin and its fermentative production
IPC: C12P19/62
Filed: 1960-01-25
Abstract: <PICT:0875011/IV (b)/1> <PICT:0875011/IV (b)/2> <PICT:0875011/IV (b)/3> A new antibiotic designated chalcomycin is produced by cultivating a chalcomycin-producing strain of streptomyces bikiensis especially S. bikiensis NRRL.2737 under aseptic aerobic conditions using submerged or surface culture techniques in an aqueous nutrient medium containing assimilable sources of carbon, nitrogen and inorganic salts. Submerged cultivation at 23-30 DEG C. and at a pH of 6-8.5 for 1-4 days with mechanical agitation is preferred. Specified sources of carbon are glucose, maltose, mannose, starch and modified starches, corn syrup, malt liquors, black-strap molasses or glycerol. Specified sources of nitrogen are amino-acids, peptones, hydrolysed and unhydrolysed proteins, fish meal, soybean meal, corn steep liquor, dehydrated corn steep liquor, meat extracts, peanut meal, inorganic nitrates, urea or ammonium salts. Inorganic salts such as sodium chloride, sodium bicarbonate, calcium carbonate or sodium nitrate may be present in the nutrient medium and also crude materials such as butanol-acetone fermentation residues, or yeast preparations. The antibiotic chalcomycin is isolated from the clarified or unclarified broth by extraction with a water-immiscible organic solvent, e.g. ethylene dichloride, chloroform, ethylacetate or amyl-acetate. The extract is purified by washing with dilute aqueous base, dilute aqueous acid and water. Further purification is obtained by (a) pouring a benzene solution of chalcomycin on an alumina column and developing and eluting the column with ethyl acetate and methanol, (b) recrystallisation of chalcomycin from tertiary butanol, (c) counter current extraction using a solvent p system consisting of n-butanol, cyclohexane and water and (d) pouring a solution of chalcomycin in aqueous acetone on an activated carbon column and developing and eluting the column with aqueous acetone and acetone. Chalcomycin has esterifiable hydroxyl groups and forms a diacetate with pyridine and acetic anhydride on catalytic hydrogenation it forms tetrahydrochalcomycin which can also be esterified and forms a diacetate. Chalcomycin is a neutral white crystalline compound, soluble in the alkyl esters of the low molecular weight fatty acids, low molecular weight alkanols, aromatic hydrocarbons, ethylene dichloride and chloroform, slightly soluble in carbon tetrachloride, water and diethyl ether and insoluble in petroleum ether, having a specific rotation [a ]27o = -43.5 (C = 1% in ethanol) a melting point on a heated block = 121-123 DEG C., containing C = 59.5-59.97%, H = 8.08-8.32% and O = 32.05-31.43% and <FORM:0875011/IV (b)/1> Me = 10.9%, having a molecular weight = 575 to 592, a U.V. spectrum as in Fig. 1, 0.00228% in absolute ethanol and I.R. spectrums in chloroform and potassium bromide as shown in Figs. 2 and 3. Chalcomycin decolourizes aqueous permanganate, adds bromine in carbon tetrachloride solution, gives a positive hydroxamate test and negative Tollen's, Iodoform and Fehlings tests. It has the properties of a lactone. On acid hydrolysis chalcomycin forms a compound melting point 187-189 DEG C., C = 60.35-60.79%, H = 7.95-8.08%, U.V. max. 308 mm . On alkaline hydrolysis chalcomycin forms a compound melting point 273-273 DEG C., C = 59.13%, H = 8.19% and O = 31.83%. Chalcomycin is active against gram positive bacteria, in particular staphylococci, pneumococci and some strains of streptococci. The antibiotic chalcomycin may be used in a pharmaceutical ointment for topical application (see Group VI).ALSO:An antibacterial ointment for topical application contains chalcomycin (see Group IV (b)) and petroleum jelly. Specified petroleum base is that having a high viscosity e.g. Jelene 50-W. The ointment contains about 0.2% by weight of chalcomycin.