Purification and characterization of aminopeptidase P from Lactococcus lactis subsp. cremoris.
Aminopeptidase P was purified 65.3-fold from the cytoplasm of Lactococcus lactis subsp. cremoris AM2 with a 5.8% yield. The purified enzyme was found to consist of one polypeptide chain with a relative molecular mass of 41,600. Metal chelating agents were found to be inhibitory and Mn2+ and Co2+ stimulated activity 7-fold and 6-fold respectively. The purified enzyme removed the N-terminal amino acid from peptides only where proline (and in one case alanine) was present in the penultimate position. No hydrolysis was observed either with dipeptides even when proline was present in the C-terminal position or when either N-terminal proline or pyroglutamate was present preceding a proline residue in the penultimate position of longer peptides. On the basis of this substrate specificity either aminopeptidase P or post-proline dipeptidyl aminopeptidase are necessary along with a broad specificity aminopeptidase to effect complete hydrolysis of casein-derived peptides containing a single internally placed proline residue. However, both aminopeptidase P and post-proline dipeptidyl aminopeptidase would be required together with a broad specificity aminopeptidase in order to completely hydrolyse casein-derived peptides that contain two internally placed consecutive proline residues. As bitter casein-derived peptides are likely to contain either single prolines or pairs of prolines, aminopeptidase P appears to be an important enzyme for debittering.
Mc Donnell
M
M
Biochemistry Department, University College Galway, Irish Republic.
Fitzgerald
R
R
Fhaoláin
I N
IN
Jennings
P V
PV
O'Cuinn
G
G
D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D002614:Chelating Agents / Q000494:pharmacology; D003035:Cobalt / Q000494:pharmacology; D003593:Cytoplasm / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D013294:Lactococcus lactis / Q000201:enzymology*; D008345:Manganese / Q000494:pharmacology; D008892:Milk / Q000382:microbiology; D008970:Molecular Weight; D010455:Peptides / Q000378:metabolism; D013379:Substrate Specificity
D002614:Chelating Agents; D010455:Peptides; D003035:Cobalt; D008345:Manganese; D000626:Aminopeptidases; C022750:X-Pro aminopeptidase