Biological and Chemical Bases for the Reclassification of Brevibacterium vitarumen (Bechdel et al.) Breed (Approved Lists, 1980) as Corynebacterium vitarumen (Bechdel et al.) comb. nov. and Brevibacterium liquefaciens Okabayashi and Masuo (Approved Lists, 1980) as Corynebacterium Ziquefaciens (Okabayashi and Masuo) comb. nov. appears on the Approved Lists, 1980 (45). The organism is of some sigdicance in the history of animal nutrition (8). Recently, it has been shown to produce corynomycolic acids linked as 6,6'-dicorynomycoloyl trehalose and 6-monocorynomycoloyl trehalose (27) and to exhibit other properties which suggest that it might belong in the genus Corynebacterium (Approved Lists, Cell walls. Cell walls were prepared by the method of Asselineau et al.(1). Acid hydrolyses were performed in sealed tubes containing 1 N HCI for sugars and 6 N HCl for amino acids and isomers of diaminopimelic acid. Hydrosoluble components of hydrolysates were studied by descending paper chromatography (Whatman no. 1) in which the following solvent systems were used: butanol-pyridine-water (6:4:3, by volume) for sugars; phenol-ammonia atmosphere for amino acids, and methanol-water-10 N HC1-pyridine (80: 17.5:2.5:10) for diaminopimelic acid (9). Sugars were visualized with p-anisidine hydrochloride and AgNOs/KOH; amino acids and diaminopimelic acid were visualized with ninhydrin. Note that the identifications of the main constituents of the cell walls of corynebacteria were performed both with cell wall preparations as described above and with delipidated cells. We found that the delipidated cells had the same composition as the cell wall preparations with regard to the main sugars, amino acids, and diaminopimelic acid. Therefore, the use of delipidated cells in lieu of cell wall preparations offers an economic and convenient means for differentiating strains quickly after lipid extraction, and the absence of free lipids makes the identification of the cell wall components easier.Guanine plus cytosine content of DNA, DNA homology, and determination of genome sizes.The preparations of corynebacterial deoxyribonucleic acids (DNAs) for determinations of the guanine plus cytosine contents and for DNA-DNA hybridizations were as previously described (6). The genome molecular weights were assessed from the product of the initial concentration of dissociated DNA (Co) and the time required for M% reassociation (the value) of the total DNA, as described by Britten and Kohne (12); see also Seidler and Mandel [43]. DNA from E.coli strain ATCC 23724 (genome molecular weight, =2.5 x 10') was used as a standard. Thus, genome molecular weight was equal to (2.5 x 109)(Cotli2 of DNA)/C,,tl,2 of E. coli DNA.
RESULTSAnalyses of the cell walls of B. vitarumen strain 12143, B. liquefaciens ATCC 14929 and two reference strains, C. diphtheriae strain HF and C. pseudodiphtheriticum strain H Toulouse, yielded galactose and arabinose as the major
INT. J. SYST. BACTERIOL.sugars and glycine, alanine, glutamic acid, and meso-diaminopimelic acid as...