The Minimal Amino Acid Requirements of Haemophilus pertussis

Abstract

SUMMARY: Of 13 sulphur-containing compounds tested only cystine, cysteine and glutathione satisfied the sulphur requirements of the strains of Haemphilus pertussis used when grown in a defined liquid medium containing 15 amino acids. Very small inocula (3-10 viable organisms) of 8 strains grew in a medium containing glutamic acid+cystine as the only amino acids added. Four strains were carried through 9 subcultures in this medium.Previous work on the sulphur requirements of Haemophilus (Bordete1la)pertussis has produced conflicting results. Different

The Growth of Bordetella pertussis: a Review

“…The effect of these factors was first tested in a medium containing charcoal-treated Casamino Acids with an inoculum of 2 x lo2 viable organisms/ml. medium, but a similar relationship was found when glutamate (with cystine) replaced the Casamino Acids, although growth was correspondingly slower (Jebb & Tomlinson, 1957). With glutamate + nicotinamide +purines +biotin + haemin, the quantity and rate of growth did not alter through two further subcultures.…”

“…Hornibrook (1939) showed that cysteine could be replaced by cystine or glutathione but not by methionine; Proom (1955) confirmed this by serial subcultivation. Cysteic acid, homocystine and sulphate are also inadequate (Jebb & Tomlinson, 1957). In experiments with large inocula, I found (Rowatt, 1957) that added cystine or cysteine increased the total crop of bacteria in media in which glutamate and yeast extract were the only source of nitrogen, i.e.…”

“…Growth was not inhibited when cysteine was autoclaved in a simple medium with glutamate as the only other amino acid (Jebb & Tomlinson, 1957).…”

“…The yield of bacteria in these two media was only 0.3 mg. dry wt./ml., possibly because these media were inadequately buffered. I have found (Rowatt, unpublished) that in a medium similar to that of Jebb & Tomlinson (1957) containing 0.01 M-glutamate and 0.1 yo (w/v) Casamino Acids, 0.3-0.5 ml. O.Ol~-alkali was formed during the growth of 0.3 mg. dry wt.…”

“…bacteria/ml., amino acids which this organism cannot synthesize would have to be added a t a concentration of approximately lO,ag./ml. Jebb & Tomlinson's (1957) medium contains 1.5 mg. starch and 1.46 mg. glutamic acid/ml., and in order to add lopug. of any other amino acid 314 E. Rowatt these compounds would have to contain 1 % of it as contaminant.…”