Simplified Approach to Identification of Aerobic Actinomycetes by Thin-Layer Chromatography

Abstract

A system has been developed for the identification of aerobic actinomycetes in the clinical laboratory based on analysis of whole cells for diaminopimelic acid and carbohydrates and on the ability of the organism to decompose casein, tyrosine, and xanthine media. The whole-cell analyses were performed by a simple thin-layer chromatographic procedure that is described. Eighteen reference cultures were correctly identified and, subsequently, 35 isolates from clinical material were grouped by using this system. The method is well suited for use in routine clinical laboratories.

The analysis of actinomycete wall amino acids by gas chromatography

“…The quantitative GC results in Table 1, however, indicate that for organisms such as Arthrobacter simplex, Mycobacterium avium, 'Nocardia' autotrophica and Nocardia asteroides alternative purification methods, perhaps using enzymes [2], are desirable. Since the derivatives used in this study did not enable the isomers of diaminopimelic acid to be separated it was necessary to screen diaminopimelic acid containing hydrolysates by TLC on cellulose plates [11]. Examined qualitatively the results obtained are, in general, comparable with published amino acid profiles ( Table 1).…”

“…The analysis of cell-wall amino acids by GC has been developed as part of an integrated small-scale chemotaxonomic procedure and uses cells from which the isoprenoid quinones and polar lipids have been removed and analysed. By providing quantitative data the present system bridges the gap between detailed peptidoglycan analyses [2] and qualitative TLC [5,11] and paper-chromatographic [4,12] studies of wall amino acids.…”

Rare actinomycete bacteria from the shallow water sediments of the Trondheim fjord, Norway: isolation, diversity and biological activity

“…While examining cultures microscopically, particular attention was paid to the formation and arrangement of spores on vegetative or/and aerial mycelium, the occurrence of special structures such as sporangia, sclerotia, synnemata, the fragmentation of mycelia, and the release of motile elements. Cell wall type was determined on the base of studying the occurrence of diaminopimelic acids and diagnostic sugars in the whole cell hydrolyzates by thin layer chromatography method (Staneck and Roberts, 1974;Hasegawa et al, 1983). On the basis of macro-and micromorphology and cell wall type, the representatives of each group were preliminary assigned to probable genera or group of genera.…”

Isoprenoid quinone, cellular fatty acid composition and diaminopimelic acid isomers of newly classified thermophilic anaerobic Gram-positive bacteria

“…Diaminopimelic acid isomers were determined by the TLC method of Staneck and Roberts [10]. Hydrochloric acid hydrolysate (10 Wl) of lyophilized cells was spotted on a cellulose TLC plate (Chromatogram-Eastman Kodak, Tokyo) or high performance cellulose TLC plate (Merck, Tokyo) together with DAP as a standard, and developed by a solvent of methanol:distilled water:6 M hydrochloric acid:…”