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The cell wall of Bacillus licheniformis N.C.T.C. 6346. Composition of the mucopeptide component.

ABSTRACT: 1. The mucopeptide component of wall preparations from Bacillus licheniformis was obtained in soluble form by treatment of the acid-insoluble residue of walls with lysozyme. 2. The soluble mucopeptide contains glutamic acid, diaminopimelic acid, alanine, N-acetylglucosamine and N-acetylmuramic acid in the molecular proportions 1.0:1.0:1.6:0.8:0.7. In addition approx. 1 mole of amide/mole of glutamic acid is present. Essentially all of the dry weight and nitrogen content of soluble mucopeptide is accounted for by these constituents. 3. The optical configurations of the amino acids were determined. Approx. 0.6 mole of d-alanine and 1.0 mole of l-alanine are present/mole of glutamic acid. 4. The structures of several small peptides derived from soluble mucopeptide after mild acid hydrolysis were established. 5. The structure of soluble mucopeptide from B. licheniformis is discussed on the basis of these results together with data on the number of free amino groups present in soluble mucopeptide.

SUBMITTER: Hughes RC

PROVIDER: S-EPMC1198467 | biostudies-other | 1968 Jan

REPOSITORIES: biostudies-other

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Project description:1. Soluble mucopeptide was prepared by lysozyme treatment of acid-extracted walls of Bacillus licheniformis N.C.T.C. 6346 and separated into fractions differing in molecular size by chromatography on Sephadex G-25 and G-50. 2. About 16% of the weight of soluble mucopeptide has a weight-average molecular weight in excess of 20000. About one half has a weight-average molecular weight of less than 2000 and the balance of soluble mucopeptide is of intermediate size. 3. In the mucopeptide fractions isolated from Sephadex there is a correlation between the weight-average molecular weight, the number of non-reducing muramic acid residues and the proportion of diaminopimelic acid residues recovered after treatment with 1-fluoro-2,4-dinitrobenzene. 4. The extent of cross-linking between peptide side chains is relatively low, even in mucopeptide material of the large molecular size. 5. The small amount of residual phosphorus present in preparations of B. licheniformis soluble mucopeptide remains associated mainly with mucopeptide material of large molecular size. 6. The mucopeptide components of lowest molecular weight are not produced as artifacts during the preparation of soluble mucopeptide, but are apparently incorporated in the insoluble mucopeptide present in walls of exponentially growing cells. 7. Soluble mucopeptide isolated in a complex with acidic polymers after lysozyme treatment of walls of B. licheniformis N.C.T.C. 6346 and Bacillus subtilis W23 retains a high molecular weight when the covalent bonds between mucopeptide and the acidic polymers are broken. 8. Pure fragments were isolated from B. licheniformis soluble mucopeptide. A major component, C1, of the material of smallest size is made up of one residue each of N-acetylglucosamine, N-acetylmuramic acid, l-alanine, glutamic acid and diaminopimelic acid. The N-acetylglucosamine is in beta-glycosidic linkage with a reducing N-acetylmuramic acid residue. The peptide unit is probably amidated. A quantitatively minor component, C2, has amino acid and amino sugar composition identical with that of component C1, but probably lacks an amide group. Another fragment, B1, is made up of two molecules of component C1 or C2 that are joined together through a molecule of d-alanine.

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The cell wall of Bacillus licheniformis N.C.T.C. 6346. Composition of the mucopeptide component.

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