FIG. 3.

Stoichiometry of the 5-oxoprolinase reaction. (A) Decyclization reaction. The reaction mixture (100 mM l-pyroglutamate and the purified enzyme in 0.1 M Tris-HCl buffer [pH 8.0]) was incubated at 30°C. At set intervals, aliquots were withdrawn and analyzed by the methods described in Materials and Methods. ○, l-glutamate; •, l-pyroglutamate. (B) Cyclization reaction. The reaction mixture (30 mM l-glutamate and the purified enzyme in 0.1 M Tris-HCl buffer [pH 8.0]) was incubated at 30°C. At set intervals, aliquots were withdrawn and analyzed by the methods described in Materials and Methods. ○, l-glutamate; •, l-pyroglutamate.