1,6-AnhMurNAc derivatives for assay development of amidase AmiD.
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.
Mercier
Frédéric
F
Centre de Recherches du Cyclotron, B30, Université de Liège, Allée du 6 août 8, Sart Tilman, B 4000 Liège, Belgium.
Zervosen
Astrid
A
Teller
Nathalie
N
Frère
Jean-Marie
JM
Herman
Raphaël
R
Pennartz
Anne
A
Joris
Bernard
B
Luxen
André
A
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
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