The Production, Assay, and Antibiotic Activity of Actidione, an Antiobiotic from Streptomyces griseus

Abstract

Actidione is a chloroform-and water-soluble antibiotic that is produced by streptornycin-yielding cultures of Streptomyces griseus and that, unlike strepto-mycin, possesses no marked antibacterial activity (Whiffen, Bohonos, Emerson, 1946). It is, however, highly active against a large number of yeasts, including the pathogen Cryptococcus neoformans. Actidione has been obtained in pure crystalline form (Leach, Ford, and Whiffen, 1947) and its empirical formula has been determined (Leach and Ford, 1948). The purpose of this paper is to report studies on the production, assay, and antibiotic activity of actidione. The production by the same organism of twvo easily differentiated antibiotics has offered an opportunity for the study of several problems of theoretical interest. It has been possible to determine the effect of changes in the composition of the fermentation medium upon the relative yields of actidione and streptomycin and to demonstrate that the amounts of the two antibiotics produced may vary independently of one another. It was of further interest to determine whether or not the loss by mutation of the ability to synthesize one antibiotic would result in an inability to synthesize the other antibiotic. PRODUCTION OF ACTIDIONE Effect of composition ofmeditum upon relative yields of streptomycin andactidione. Actidione was produced in 100-ml aliquots of medium contained in 500-ml Erlenmeyer flasks that were incubated in a reciprocal shaker at 24 C. The flasks were inoculated with 5 ml each of a 72-hour-old vegetative growth of S. griseus, phage-resistant strain RM-241CR (Saudek and Colingsworth, 1947). Assays for actidione were performed by a paper-disk plate method that is to be described later. Streptomycin was assayed by the method of Loo et al. (1945). The yields of actidione and streptomycin were first compared in two different media, identified as no. 25 and no. 26. Medium 25 contained the following: "cerelose" 20.0 g, KCl 4.0 g, CaCO3 8.0 g, (NH4)2SO4 5.0 g, dried brewers' yeast (Standard Brands no. 2019) 2.5 g, KH2PO4 0.2 g, distilled water 1,000 ml. In this medium high yields of streptomycin were obtained, but only small amounts of actidione wvere produced (table 1). On the other hand, the yield of actidione wvas superior to that of streptomycin in medium 26, wvhose composition was as follows: cerelose 10.0 g, soybean oil meal 10.0 g, "curbay B-G" (a distiller's soluble from U. S. Industrial Alcohol) 5.0 g, CaCO3 1.0 g, NaCl 5.0 g, distilled water 1,000 ml (table 1). 283