title: 1,6-AnhMurNAc derivatives for assay development of amidase AmiD.
abstract: Various peptidoglycan fragments were synthesized from two anhydro-muramic acid derivatives protected with a Bn or a PMB group at the 4th position, in homogenate phase or on a solid support. In order to facilitate HPLC detection, a chromophoric group was attached to the peptide chain. The periplasmic amidase sAmiD of Escherichia coli was used to cleave the amide bond between the lactyl group of the MurNAc and the α-amino group of L-Ala where the peptide chain was at least a dipeptide (L-Ala-γ-D-Glu) amidated by benzylamine on the γ-carboxyl group of D-Glu. In the presence of a tripeptide chain (L-Ala-γ-D-Glu-L-Lys) or a tetrapeptide chain (L-Ala-γ-D-Glu-m-A(2)pm-D-Ala) higher hydrolysis rates were observed. We have also demonstrated that the presence of TNB on the ε-amino group of L-Lys only has a small influence on the hydrolysis capacity of sAmiD.
authors.affiliation: Centre de Recherches du Cyclotron, B30, Université de Liège, Allée du 6 août 8, Sart Tilman, B 4000 Liège, Belgium.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Mercier
authors.forename: Astrid
authors.initials: A
authors.lastname: Zervosen
authors.forename: Nathalie
authors.initials: N
authors.lastname: Teller
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Frère
authors.forename: Raphaël
authors.initials: R
authors.lastname: Herman
authors.forename: Anne
authors.initials: A
authors.lastname: Pennartz
authors.forename: Bernard
authors.initials: B
authors.lastname: Joris
authors.forename: André
authors.initials: A
authors.lastname: Luxen
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000378:metabolism; D001596:Benzylamines / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D009112:Muramic Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology
chemical_list: D001596:Benzylamines; D029968:Escherichia coli Proteins; D009112:Muramic Acids; C555722:N-acetyl-1,6-anhydromuramyl-alanyl-gamma-glutamyl-(N-epsilon-(2,4,6-trinitrophenyl)lysine); D009842:Oligopeptides; C430327:anhydro-N-acetylmuramic acid; C030796:benzylamine; D000581:Amidohydrolases; C019665:amidase