Data for the synthesis of oligo-gamma-glutamylglutamines as model compounds for gamma-glutamyltransferases (GGTs) and for normalization of activities of different GGTs

gamma-Glutamyltransferases (GGTs) are widespread, conserved enzymes that catalyze the transfer of the gamma-glutamyl moiety from a donor substrate to water (hydrolysis) or to an acceptor amino acid (transpeptidation) through the formation of a gamma-glutamyl enzyme intermediate. Although the vast majority of the known GGTs has a short sequence called lid-loop covering the glutamate binding site, B. subtilis GGT and some other enzymes from Bacillus spp. lack the lid loop. In order to assess the possible role of the lid loop of GGTs in substrate selection, synthetic oligo-gamma-glutamylglutamines containing up to three gamma-glutamyl residues were used as model substrates. The activities of the enzymes under investigation were standardized with respect to a common reaction to ensure comparable results. The activity of an engineered mutant enzyme containing the amino acid sequence of the lid loop from E. coli GGT inserted into the backbone of B. subtilis GGT was compared to that of the lid loop-deficient B. subtilis GGT and the lid loop-carrier E. coli GGT (Calvio, Romagnuolo, Vulcano, Speranza, Morelli Enz. Micr. Technol. 2018 [1]). Here we report the experimental procedures for the synthesis of model substrates gamma-glutamylglutamines through the method of the N-phtaloyl-L-glutamic acid anhydride and the spectral data of the synthetized compounds. The data obtained in the normalization procedure of the activities of the three enzymes are also reported.