Bluensomycin was obtained from cultures of Streptomyces verticillus, or the
same substance produced by any other means. For example antibiotic was
prepared by growing of Streptomyces bluensis NRRL 2876 biological way and
isolation from cultural solution by adsorption with a cation-exchange resin or a
capillary adsorption method by elution with water-acid solution at pH from 1
to 6 or acidic water solution of acetone.
5350 L of cultivating liquid with pH 8.2 was mixed with 16 kg oxalic acid
acidified with 1 N sulfuric acid to pH 2.9 and was filtered through about 160
kg fossil flour and washed with 500 L water. The filtrate (about 5400 L) was
alkalified to pH 7.8-8 with 10% sodium hydroxide and was filtered through
fossil flour filter. Then it was passed through two column with polyacrylic acid
cation exchange resin in sodium form (US Patent No. 2,915,432).
Each column was 35 cm in diameter and contained 0.126 kg of above resin.
The filtrate (5300 L) was passed with rate 19 L/minute. Then the columns
were washed with deionized water, 1 N sulfuric acid to pH 1.2-1.5, and at last
eluted with 4x100 L water. About 200 L of column effluent was alkalifed to pH
6.4 with 10% sodium hydroxide. The 1-st column effluent was mixed with
1200 g of activated carbon, the second effluent was mixed with 850 g of coal
(1 g coal per 1 g dissolved product). The mixture was thoroughly stirred and
filtered. Each coal precipitate was washed 3x10 L with water and 200 L 15%
water acetone.
Water acetone effluent from the 1-st column (187 L) was dried and gave 1034
g of bluensomycin, the second gave 777 g. The portions of antibiotic were
combined and purified by chromatography. The column (high 1.2 m, volume
155 L, with sea sand, adsorbent cotton and fossil flour as the carrier) was
used. It was washed with 150 L of deionized water and 300 L 10% water
acetone (rate 410 ml per minute). A fraction 101-127 L water acetone gave
640 g bluensomycin after drying. The IR and UV spectra, element analysis
confirmed the structure of prepared product and its purity.
