Production and purification of bacilysin.
1. Bacilysin, a hydrophilic substance formed by certain aerobic spore-forming bacteria that causes lysis in cultures of growing staphylococci, has been produced in aerated cultures of a strain of Bacillus subtilis (A14). A chemically defined medium was used, which contained glucose, Czapek-Dox salts and ferric iron. Production of bacilysin occurred, after a lag, while the culture was still undergoing rapid growth. 2. Bacilysin was adsorbed from the culture medium on Zeo-Karb 225 (SR5) (H(+) form) and eluted with aqueous pyridine. The crude material was purified by chromatography in pyridine-acetate buffers on columns of Dowex 50 (X2) and Dowex 50 (X8) respectively and by chromatography in aq. 70% (v/v) propan-2-ol on Sephadex G-25. 3. Purified bacilysin behaved as a single ninhydrin-positive substance when subjected to chromatography on paper in butan-1-ol-acetic acid-water and to electrophoresis on paper at pH4.5 or pH1.8. At pH4.5 the substance behaved as though it had no net change and at pH1.8 it migrated towards the cathode.
Rogers
H J
HJ
Sir William Dunn School of Pathology, University of Oxford.
Newton
G G
GG
Abraham
E P
EP