A growth factor for C. diphtheriae present in liver

Abstract

KETONIC ACIDS DERIVED FROM CHOLIC ACID 111 washed with water. This gum gave an H-test showing yellow-greenish-violet (dichroic) colours. It was dissolved in acetic acid (10 ml.) and the cold solution treated with 20% CrO (1 ml.). After 15 min. it was diluted: the partly crystalline precipitate was collected after 2-3 hr., washed, and recrystallized from dilute ethanol. Yield, 0 55 g. of needles, m.p. 185B188°, raised to 190-191°by recrystallization from l.p./benzene and ethyl acetate. (Found: C, 69 3; H, 8*8. Calc. for C29H,o07: C, 69*0; H, 8.8%.) H-test, negative. This ester (0.3 g.) was heated in a sealed metal bomb for 4 hr. at 200-210O with a solution of NTa (0.1 g.) in ethanol (2 ml.) with hydrazine hydrate (0.3 ml. of 95-99%). Acidification of the diluted contents of the cooled bomb precipitated a solid which separated from dilute ethanol as long white needles (0.15 g.) of 7:12-dihvdroxy-cholanic acid (III), m.p. 205°. SUMMARY 1. The series of six% possible acids obtainable by the oxidation to carbonyl of one or two secondary hydroxyl groups in natural cholic acid has been completed by the preparation of 12-hydroxy-3:7diketo-cholanic acid. 2. New derivatives of this acid, of 3:12-dihydroxy-7-keto-, 7:12-dihydroxy-3-keto-, and 3-hydroxy-7:12-diketo-cholanic acids are described; methods for obtaining such substances have been further explored. The author thanks Allen and Hanburys Ltd. for a gift of cholic acid.

Growth factors for Corynebacterium diphtheriae. 4. Preparation from yeast

“…Basal medium. The basal medium employed, shown in Table 1, differed from that described by Chattaway et al (1944). Certain amino-acids hitherto omitted were included in order to bring its composition more nearly into line with a casein-hydrolysate medium on which the organism grows profusely.…”

“…Dolby, Happold & Sandford (1944), but showed some similarity to strepogenin, which was prepared by Woolley (1941) from various purified proteins, and is required by certain strains of Streptococcus haemolyticus. The present paper reports the separation and concentration from yeast of a number offactors similar in many respects to the liver factor of Chattaway et al (1944).…”

“…45,584. Growth Factors for Corynebacterium Diphtheriae 4. PREPARATION FROM YEAST BY F. W. CHATTAWAY, DORIS E. DOLBY, D. A. IIALL ANm FRANK C. HAPPOLD Department of Biochemi8try, Univer8ity of Leeds (Received 15 February 1949) In an earlier paper, Chattaway, Happold & Sandford (1944) showed that certain gravis (and intermedius) strains of Corynebacterium diphtheriae required an unidentified growth factor when cultivated on a medium of known composition containing pimelic, nicotinic and pantothenic acids (Mueller, 1937 a, b;Evans, Handley & Happold, 1939a, b). This factor, present in liver, was shown to be dialysable and resistant to proteolytic enzymes.…”

Unidentified Members of the Vitamin B Complex

“…'The work began by the recognition by IIvans, Handley and Happold (1939) of a factor required by certain strains of C'oryzebucterium diphtheriae intermedius and the concurrent investigation into growth factors required by the rat and present in the same liver fractions (Hind et al 1939;Lythgoe et al 1940). This factor has been concentrated and been shown to be distinct from a factor or factors required by L. casei E and also present in the liver extracts, when this organism is grown on a casein hydrolysate medium (Chattaway, Ihppold, Lythgoe, Sandford & Todd, 1943 ;Chattaway, Happold & Sandford, 1944). But it has also been demonstrated (Dolby, Happold & Sandford, 1944) that svhen L. cnsei E is grown on an amino-acid medium that this C. diphtheriue factor is essential for its growth.…”

Growth Factors for Corynebacterium diphtheriae Strain Dundee

“…Chattaway, Happold & Sandford (1944) and Chattaway, Dolby, Hall & Happold (1949) reported the presence in liver and yeast concentrates of a factor or factors essential for the growth of certain intermediate and gravis strains of Cwynebucterium dipintherim on an otherwise chemically defined medium. Working with acid hydrolysates of brewers' yeast they obtained chromatographic data which suggested that the active material was associated with four ninhydrin-reacting substances.…”