Autolysis of isolated cell walls of Bacillus licheniformis N.C.T.C. 6346 and Bacillus subtilis Marburg Strain 168. Separation of the products and characterization of the mucopeptide fragments.
1. Cell walls were isolated from Bacillus licheniformis N.C.T.C. 6346 and Bacillus subtilis Marburg strain 168 trp grown on casein hydrolysate into exponential phase. Autolysis was carried out and the soluble products, separated by chromatography on DEAE-cellulose, from the two wall preparations are broadly similar in composition and are in agreement with autolysis proceeding with hydrolysis of amide bonds between l-alanine and N-acetylmuramic acid residues in the mucopeptide components. 2. Peptides originating from the mucopeptide components were isolated and shown to be a monomer peptide, l-alanyl-d-glutamyl-meso-diaminopimelic acid and a dimer peptide containing two monomer peptides linked through a residue of d-alanine. Approximately one amide group is present for each equivalent tripeptide unit and is probably substituted on diaminopimelic acid residues. 3. Oligosaccharides originating from the mucopeptide components were isolated and after hydrolysis contained almost equimolar amounts of glucosamine and muramic acid and only very small amounts of amino acids. The number-average chain length, estimated by the release of non-reducing end groups of N-acetylglucosamine with exo-beta-N-acetylglucosaminidase, is approximately ten hexosamine residues for oligosaccharides isolated from either organism. The oligosaccharides are polydisperse. 4. N-Acetylglucosamine residues are the only reducing terminals detectable in the oligosaccharides isolated from B. subtilis or B. licheniformis cell-wall autolysates. The number-average chain lengths of the oligosaccharides were determined by estimation of the content of these residues and are higher than those found by enzymic assay. Possible reasons for the discrepancy are discussed.
D001329:Autolysis; D001407:Bacillus / Q000378:metabolism*; D001412:Bacillus subtilis / Q000378:metabolism*; D001896:Boron Compounds; D002473:Cell Wall / Q000032:analysis / Q000378:metabolism; D002848:Chromatography, DEAE-Cellulose; D002854:Chromatography, Paper; D004586:Electrophoresis; D005944:Glucosamine / Q000032:analysis; D006026:Glycoside Hydrolases; D006595:Hexosamines / Q000032:analysis; D009088:Mucoproteins / Q000032:analysis*; D009844:Oligosaccharides / Q000032:analysis; D010084:Oxidation-Reduction; D010455:Peptides / Q000032:analysis; D010504:Periodic Acid
D001896:Boron Compounds; D006595:Hexosamines; D009088:Mucoproteins; D009844:Oligosaccharides; D010455:Peptides; D010504:Periodic Acid; D006026:Glycoside Hydrolases; D005944:Glucosamine