PRODUCTION, PURIFICATION, AND CHARACTERIZATION OF SYNNEMATIN

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Abstract
In an Emericellopsis terricola var. glabra fermentation, a medium containing 4% glucose, 1% CaCO3, 40% soybean meal extract, 0.1% (NH4)2SO4 and 5 ml of salt mixture per 50 ml was found suitable for synnematin production. Intermittent addition of one of several oils stimulated the production of synnematin greatly. Highest yields (888 units per ml; roughly, 1 g/l) were obtained by feeding cottonseed oil to the soybean meal extract medium. When DL-[alpha]-aminoadipic acid and DL-diaminopimelic acid were tried as precursors for synnematin production, no significant increase in yield was observed. When L-lysine was used, the yield of synnematin increased from 810 to 1201 units per ml. The optimal pH for synnematin production on the natural medium was between 6.0 and 6.5. When L-lysine or DL-lysine was fed to the fermentation, 50 to 60% increase in yield was obtained at the same pH plateau (6.0 to 6.5) as a control. Therefore, it was concluded that L-lysine and DL-lysine stimulated synnematin, production, probably acting as precursor. An attempt was made to purify S35-synnematin B produced when L-lysine was fed as precursor and 1.0 mc of -S35O4 added to each fermentation. A combination of a charcoal elution column and a Celite 545 partition column was successful in isolating S35-synnematin B, entirely free from S35-contaminants. Synnematin B preparations of more than 95% purity (maximum purity, 99.7%) were prepared in this way. Acid hydrolysis of pure synnematin B liberated [alpha]-aminoadipic acid and [beta]-thiovaline. [alpha]-Aminoadipic acid was isolated in a crystalline form. It can be concluded that the antibiotic produced by E. terricola var. glabra, growing on a soybean meal extract medium, with intermittent feeding of cottonseed oil and L-lysine, is identical with synnematin B.