Vol. 70 from 250 cc. of hot ethanol. The yield of pure product was 1 g., m. p. 185-188°dec.Anal. Calcd. for C22H39O6NS: N, 3.14. Found: N (Dumas), 2.84, basic N (perchloric acid titration in acetic acid), 2.76.The product was soluble in sodium bicarbonate solutions and in hot dilute hydrochloric acid solution. Alkaline titration indicated the presence of one carboxyl and one lactone group. The reaction product obtained from cysteine and Afl.r-angelicalactone11 when titrated with perchloric acid showed the absence of a basic amino group; protolichesterinic acid appears to have added cysteine through the -SH group without secondary involvement of the amino group. Antibacterial Tests.-Tests against the Streptococcus, Staphylococcus and Bacillus typhi were conducted in tryptose phosphate medium. The inoculum was a 1:1000 dilution of a twenty-four-hour culture of the organism and incubation was for eighteen hours at 37°. The anaerobe was tested in Bacto-Anaerobe Medium with Dextrose. The acid-fast organisms, B. tuberculosis ranae and the human tuberculosis strain H37Rv were grown in submerged culture in Youmans' modification14 of Proskauer-Beck synthetic medium. The inoculum with ranae was a (14) Composition: Asparagin, 0.5%; primary potassium acid phosphate, 0.5%, potassium sulfate, 0.05%; magnesium citrate, 0.15%; and glycerol, 2% in distilled water.1:100 dilution of a forty-eight hour culture in Long's synthetic medium; incubation was for forty-eight hours at 37°. An inoculum of 0.02 mg. of fresh bacteria per cc. of test medium was used with H37Rv; incubation at 37°w as for fourteen days, excellent growth being evidenced after eight days.Acknowledgment.-We are indebted to M. E. Auerbach and Staff for analytical data, to Dr. F. C. Nachod for surface-tension measurements, and John W. Klimek for antifungal tests.
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