Prolidase activity ofLactococcus lactissubsp.cremorisAM2: partial purification and characterization

Summary Prolidase activity from cytoplasm ofLactococcus lactissubsp.cremoris (Streptococcus cremoris)AM2 was partially purified. The enzyme hadMr42000 and optimum activity between pH 7·35 and 8·25 in citrate, phosphate and borate buffers while in a universal buffer system an optimum pH between 8·3 and 9·0 was observed. The activity was strongly inhibited by the chelating agents E.DTA, 1,10-phenanthroline and 8-hydroxyquinoline. Inhibition was also noted with dithio-threitol,N-ethylmaleimide and bacitracin. The enzyme was active on all amino-acylproline substrates tested except Gly-Pro and Gip-Pro and also showed activity against Pro-Pro. While most prolyl amino acids tested were not hydrolysed, hydrolysis was noted with Pro-Ala and Pro-Val.Kmvalues of 20 mM and 10 mM were obtained with Phe-Pro and Met-Pro respectively; however, substrate inhibition was observed with Ile-Pro and Leu-Pro.