Enzymatic synthesis of chromogenic substrates for Glu,Asp‐specific proteinases

Abstract

Glu,Asp-specific endopeptidases represent a new subfamily of chymotrypsin-like proteolytic enzymes. These enzymes prefer Glu or Asp residues in the P1 position of the substrates. p-Nitroanilides of N-acylated di-, tri- and tetrapeptides with C-terminal glutamic or aspartic acid residues have been obtained. Acyl peptide p-nitroanilides were synthesized via acylation of glutamic or aspartic acid p-nitroanilides using methyl esters of the respective N-acylated peptides, generally with good yields. The reactions were performed in organic solvents using subtilisin 72 sorbed on silica as a catalyst. The kinetic parameters for the hydrolysis of these p-nitroanilides with proteinases from Bacillus intermedius and Bacillus licheniformis were determined.

SP-1, a Serine Protease from the Gut Microbiota, Influences Colitis and Drives Intestinal Dysbiosis in Mice

“…As indicated, in position P1, both Arg and Lys were preferred over other residues (Figure 3a). Note that most subtilisin-like proteases reportedly do not cleave substrates with basic residues at this position and often show a preference for Glu, Asp, Phe, and Leu [55,56]. This result is, though, in agreement with data provided by Gosalia et al of a subtilisin Carlsberg accepting P1 basic residues [57].…”

Expression, purification, and characterization of soluble and active glutamate-specific endopeptidase in Bacillus licheniformis and Pichia pastoris

“…GSE-BL exhibits much higher specificity and catalytic efficiency toward Glu residue than glutamate-specific endopeptidase from other microorganisms, including Streptomyces fradiae [3], Staphylococcus aureus [4], and Bacillus subtilis [5]. GSE-BL is broadly applied in peptide synthesis [6], bio-nano-material preparation [7], protein sequence analysis [8], and peptide and protein purification because of its high specificity and catalytic efficiency [9,10].…”