CHEMISTRY OF PHOSPHATIDE AND WAX D OF MYCOBACTERIUM TUBERCULOSIS BCG IN ONE WEEK CULTURE

Abstract

“Purified wax” as isolated by Anderson (1929), is a complex lipid extracted with chloroform from cultures of acid fast bacilli. Extracting the purified wax with boiling acetone, Asselineau (1951) obtained a soluble portion termed “Wax C” and an insoluble residue, the “Wax D” fraction. Likewise, the alcohol-ether soluble part of acid fast bacilli was classified into several fractions. “Phosphatide” is a boiling acetone insoluble residue in the alcohol-ether soluble fraction, corresponding to Wax D of the alcohol-ether insoluble and chloroform soluble fraction. Therefore, the substances contained in Wax D can be migrated from “phosphatide” depending on the conditions of extraction and on the metabolic stage of bacterial cells.
Works in this laboratory elucidated the chemical components of Wax A, C (Tsumita, 1956 a, b) and Wax D (Nojima, 1959 a, b) of BCG in 4-6 weeks culture. In this experiment a special consideration was paid on the migration of substances in Wax D from “phosphatide” depending on the extracting condition and on the age of the culture. Therefore, the present author intended to compare the yield and some chemical natures of “phosphatide” with those of Wax D in one week as well as in 4 weeks culture of the bacterial cells. The data of the latter were already precisely reported though its chromatographic fractionation has not been described (Nojima et al., 1958) .
Substances containing phosphorus in “phosphatide” and Wax D will play an important role in the metabolic process of lipid in Mycobacterium. After chromatographic partitioning of these fractions phosphorus and inositol contents were analyzed and compared. On the other hand, special emphasis was placed on the yield of cord factor contained in Wax D depending on the age of the culture.