Literature summary for 3.4.11.5 extracted from

Wang, K.D.; Wang, K.H.; Zhou, N.D.; Tian, Y.P.
Secretory expression, purification, characterization, and application of an Aspergillus oryzae prolyl aminopeptidase in Bacillus subtilis (2017), Appl. Biochem. Biotechnol., 181, 1611-1623 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene pap, recombinant expression of N-terminal His6-tagged enzyme in Bacillus subtilis strain WB600, the enzyme is secreted by adding 2 mM CaCl2 and 5% D-sorbitol, extracellular and intracellular PAP activities are 7.2 and 78.6 U/ml, respectively. Sorbitol and mannitol raise the level of the secretion, but sorbitol is slightly better than mannitol and increases 100.19% compared with the control. CaCl2 can further improve the secretion level of the extracellular enzyme	Aspergillus oryzae

Cloned (Comment)

Organism

gene pap, recombinant expression of N-terminal His6-tagged enzyme in Bacillus subtilis strain WB600, the enzyme is secreted by adding 2 mM CaCl2 and 5% D-sorbitol, extracellular and intracellular PAP activities are 7.2 and 78.6 U/ml, respectively. Sorbitol and mannitol raise the level of the secretion, but sorbitol is slightly better than mannitol and increases 100.19% compared with the control. CaCl2 can further improve the secretion level of the extracellular enzyme

Aspergillus oryzae

Inhibitors

Inhibitors	Comment	Organism	Structure
Cu2+	strong inhibition	Aspergillus oryzae
Zn2+	strong inhibition	Aspergillus oryzae

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.054	-	Pro-4-nitroanilide	pH 7.5, 50°C, recombinant enzyme	Aspergillus oryzae

Metals/Ions

Metals/Ions	Comment	Organism	Structure
NaCl	activates at up to over 4 M	Aspergillus oryzae

Organism

Organism	UniProt	Comment	Textmining
Aspergillus oryzae	W8GG09	-	-
Aspergillus oryzae JN-412	W8GG09	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminal His6-tagged PAP 4.3fold from Bacillus subtilis by dialysis, nickel affinity chromatography, and ultrafiltration	Aspergillus oryzae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	the enzyme cleaves N-terminal Pro residues from many peptides but shows varying hydrolysis rates for various Pro-X dipeptides or peptides of different lengths. The recombinant prolyl aminopeptidase hydrolyzes Pro-4-nitroanilide specifically and no activity is observed toward other 4-nitroanilide substrates. No activity with Leu-Pro. Enzyme PAP can act on casein	Aspergillus oryzae	?	-	?
additional information	the enzyme cleaves N-terminal Pro residues from many peptides but shows varying hydrolysis rates for various Pro-X dipeptides or peptides of different lengths. The recombinant prolyl aminopeptidase hydrolyzes Pro-4-nitroanilide specifically and no activity is observed toward other 4-nitroanilide substrates. No activity with Leu-Pro. Enzyme PAP can act on casein	Aspergillus oryzae JN-412	?	-	?
Pro-4-nitroanilide + H2O	-	Aspergillus oryzae	Pro + 4-nitroaniline	-	?
Pro-Leu + H2O	69.5% activity compared to Pro-Phe-Gly-Lys	Aspergillus oryzae	Pro + Leu	-	?
Pro-Leu + H2O	69.5% activity compared to Pro-Phe-Gly-Lys	Aspergillus oryzae JN-412	Pro + Leu	-	?
Pro-Leu-Ser-Arg-Thr-Leu-Ser-Val-Ala-Ala-Lys-Lys + H2O	17.4% activity compared to Pro-Phe-Gly-Lys	Aspergillus oryzae	Pro + Leu-Ser-Arg-Thr-Leu-Ser-Val-Ala-Ala-Lys-Lys	-	?
Pro-Lys + H2O	74.4% activity compared to Pro-Phe-Gly-Lys	Aspergillus oryzae	Pro + Lys	-	?
Pro-Lys + H2O	74.4% activity compared to Pro-Phe-Gly-Lys	Aspergillus oryzae JN-412	Pro + Lys	-	?
Pro-Phe-Gly-Lys + H2O	best peptide substrate	Aspergillus oryzae	Pro + Phe-Gly-Lys	-	?
Pro-Phe-Gly-Lys + H2O	best peptide substrate	Aspergillus oryzae JN-412	Pro + Phe-Gly-Lys	-	?
Pro-Pro + H2O	44.1% activity compared to Pro-Phe-Gly-Lys	Aspergillus oryzae	Pro + Pro	-	?
Pro-Pro + H2O	44.1% activity compared to Pro-Phe-Gly-Lys	Aspergillus oryzae JN-412	Pro + Pro	-	?

Subunits

Subunits	Comment	Organism
?	x * 50000, recombinant His-tagged enzyme, SDS-PAGE	Aspergillus oryzae

Synonyms

Synonyms	Comment	Organism
PAP	-	Aspergillus oryzae
Prolyl aminopeptidase	-	Aspergillus oryzae

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	recombinant His-tagged enzyme	Aspergillus oryzae

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
50	-	purified recombinant His-tagged enzyme, stable below	Aspergillus oryzae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	recombinant His-tagged enzyme	Aspergillus oryzae

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
6	11	purified recombinant His-tagged enzyme, stable within this range	Aspergillus oryzae