title: Investigations of the interactions of saccharides with the lysozyme from bacteriophage lambda.
abstract: The bacteriophage lambda R gene has been isolated into an Escherichia coli expression system and the R gene product, a lysozyme, has been overexpressed and purified to homogeneity using an efficient purification procedure. A turbidimetric assay utilizing chloroform-treated E. coli cells has been optimized to assess the bacteriolytic activity of the purified enzyme. Using this assay, oligomers of beta (1 --> 4) N-acetyl-D-glucosamine at high concentrations were shown to inhibit lysozyme but were not cleaved by the enzyme. Differential scanning calorimetry revealed that the thermal denaturation of lysozyme was found to increase in the presence of (GlcNAc)3 and (GlcNAc)5. The lysozyme was also expressed in an E. coli strain auxotrophic for methionine, allowing for the incorporation of [methyl-13C]methionine into the enzyme. An alteration of the [1H-13C]HMQC NMR spectra of the labelled enzyme was observed in the presence of (GlcNAc)5. Commercially available nitrophenyl glycosides did not act as substrates for lambda lysozyme. The results indicate that lambda lysozyme has specific interactions with oligosaccharides of N-acetylglucosamine, but is incapable of hydrolyzing these sugars. The relevance of the structure of peptidoglycan to the activity of lambda lysozyme is discussed.
authors.affiliation: Department of Chemistry, University of Waterloo, Ont. Canada.
authors.forename: H S
authors.initials: HS
authors.lastname: Duewel
authors.forename: E
authors.initials: E
authors.lastname: Daub
authors.forename: J F
authors.initials: JF
authors.lastname: Honek
mesh_terms: D010582:Bacteriophage lambda / Q000201:enzymology*; D002240:Carbohydrate Sequence; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry* / Q000302:isolation & purification; D009844:Oligosaccharides / Q000737:chemistry*; D010457:Peptidoglycan / Q000737:chemistry
chemical_list: D009844:Oligosaccharides; D010457:Peptidoglycan; D009113:Muramidase