微生物の生産する抗菌物質(第1報) : 放線状菌1005號菌の生産する抗菌物質

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We have isolated 10 strains of actinomyces which produces antibiotic substance from soil. Among them the antibiotic substance produced by strain No. 110 and No. 1005 have been investigated most closely. Strain No. 1005 produced antibiotic substance chemically resembling Streptothricin and Streptomycin, and No. 110 to Actinomycin A.This strain was newly isolated from soil by enrichment method. Its characteristic are as follows : -i) Filament, fine straight, branching, with no spiral ; conidia abundant, oval.ii) Gelatin stab : liquefied gradually.iii) Krinsky agar : growth orange, aerial mycellium in four to seven days, at first white then purple, at last brownish.iv) Starch, Lactose, Dextrin agar containing pepton : like Krinsky agar, but produceing brown pigment.v) Potato : under the colony potato becomes brown, then blackish.vi) Optimum temperature : 30°〜37℃.This strain very closely resembles A. lavendualc.(2) Necessary conditions for the production of antibiotic substance.i) By surface culture.a) Culture temperature…26℃〜28℃.b) Culture media.…Starch 1%, pepton 0.5%, K_2HPO_4 0.2%, NaCl 0.2%, FeSO_4・7H_2O 0.001%, PH 7.0.This strain produces the antibiotic substance in highest quantity in 8〜14 day culture and inhibits St. aureus. Terajima strain at a dilutien of 1 : 10000 and E. coli. comnnis at a dilution of 1 : 20.ii) By shake culture.The highest potency was obtained in 3〜4 day culture by this method, but the potency was not promoted, 3) Purification of the antibiotic substance.The culture fluid is adjusted to PH 4,by adding dilute Hcl, and is filtered. Then the filtrate is absorbed by charcoal (1%) at PH 7.0 and eluted from the charcoal into acid (PH 2.0) methanol (95%) by heating at 50℃ for an hour. Then the methanol is distilled off under vacum at 45〜50℃, and brown water solution is left. Aceton added to it, precipitates is the active element. This precipitate is slowly dissolved in methanol by adding HCl drop by drop until PH becomes 2.0. Then from this acid methanol solution the active element is precipitated by adding aceton. The yield is 10〜30%. (4) Properties of the antibiotic substance. The powder is considerably hygroscopic, easily soluble in water and in acid methanol, but not in aceton, ether and other organic solvents. The dilution and cup method arc used for the potency test of this anribiotic substance. The antibacterial activity depends considerably upon the PH and NaCl composition of the test broth. The antibiotic substance in culture fluid is thermostable, for it is scarcely destroyed even when heated at 80℃ for 30 minutes at PH 2. It has bacteriostatic and bactericidal action. In animal experiments, the M. L. D. of the powder is about 15mg/10g against mice, so it is scareely toxic. Its antibacterial activity is as follows.[table]
1949-04-15

微生物の生産する抗菌物質(第1報) : 放線状菌1005號菌の生産する抗菌物質

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