Some Enzymatic Properties and Substrate Specificities of Peptidoglutaminase-I and II

Some enzymatic properties and substrate specificities of the two peptidoglutaminases (peptidoglutaminase-I (PGase-I) and II (PGase-II) isolated from Bacillus circulans were investigated.

The both resembled each other with respect to optimum pH and temperature for their activities, susceptibility to various reagents and effects of many metal ions. But, the pH-rate profile of PGase-II was more broad than that of PGase-I. Thermal stability of PGase-I was better than that of PGase-II at a degree of about 5°C.

However, they were definitively different each other with respect to their substrate specificities. PGase-I specifically deamidated the γ-amide of L-glutamine residing at the carboxyl terminal on peptides, and was inactive to glutamine derivative substituted at both α-amino and α-carboxyl groups. On the other hand, the best substrate for PGase-II was tri-peptides, X-Gln-Y, where X was carbobenzoxy-,t-amyloxycarbonxy- or amino acid residues, and Y was amino acids. Though L-glutamine presented in polypeptide chains composed of more than four amino acid residues was a poor substrate, two L-glutamines in oxidized insulin A chain were well attacked by the enzyme.

The both PGase were inactive to asparagine and asparaginyl peptides.