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Literature summary for 3.4.11.7 extracted from

  • Ewert, J.; Glueck, C.; Strasdeit, H.; Fischer, L.; Stressler, T.
    Influence of the metal ion on the enzyme activity and kinetics of PepA from Lactobacillus delbrueckii (2018), Enzyme Microb. Technol., 110, 69-78 .
    View publication on PubMed

Application

Application Comment Organism
synthesis the enzyme is interesting for an industrial application, because of the high specificity for N-terminal Asp and Glu Lactobacillus delbrueckii subsp. lactis

Cloned(Commentary)

Cloned (Comment) Organism
gene pepA, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Lactobacillus delbrueckii subsp. lactis

Inhibitors

Inhibitors Comment Organism Structure
acetic acid inactivates at 50% w/v Lactobacillus delbrueckii subsp. lactis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics Lactobacillus delbrueckii subsp. lactis

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates Lactobacillus delbrueckii subsp. lactis
Cu2+ activates Lactobacillus delbrueckii subsp. lactis
Mn2+ activates Lactobacillus delbrueckii subsp. lactis
additional information PepA ia a metallopeptidase. The metal center can be described as [M1M2-enzyme], where M1 and M2 indicate the type of metal ion in site 1 and site 2. The activity and reaction kinetics, as well as the pH and temperature profile differ depending on the metal ion used in the reaction or reactivation. Quantification of activity of reactivated recombinant apo-PepA with different metal ions, overview Lactobacillus delbrueckii subsp. lactis
Ni2+ activates Lactobacillus delbrueckii subsp. lactis
Zn2+ activates Lactobacillus delbrueckii subsp. lactis

Organic Solvent Stability

Organic Solvent Comment Organism
acetic acid inactivates at 50% w/v Lactobacillus delbrueckii subsp. lactis

Organism

Organism UniProt Comment Textmining
Lactobacillus delbrueckii subsp. lactis F0HXE4
-
-
Lactobacillus delbrueckii subsp. lactis DSM 20072 F0HXE4
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-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by dialysis, nickel affinity chromatography, and desalting gel filtration Lactobacillus delbrueckii subsp. lactis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.85
-
recombinant reactivated CuCu-PepA, pH 7.0, 55°C, substrate L-aspartate-beta-naphthylamide Lactobacillus delbrueckii subsp. lactis
1.3
-
recombinant reactivated NiNi-PepA, pH 6.0, 60°C, substrate L-aspartate-beta-naphthylamide Lactobacillus delbrueckii subsp. lactis
27.2
-
recombinant reactivated ZnZn-PepA, pH 6.0, 60°C, substrate L-aspartate-beta-naphthylamide Lactobacillus delbrueckii subsp. lactis
53.3
-
recombinant reactivated CoCo-PepA, pH 6.0, 65°C, substrate L-aspartate-beta-naphthylamide Lactobacillus delbrueckii subsp. lactis
78.6
-
recombinant reactivated MnMn-PepA, pH 6.0, 60°C, substrate L-aspartate-beta-naphthylamide Lactobacillus delbrueckii subsp. lactis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Asp-Ala + H2O
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Lactobacillus delbrueckii subsp. lactis Asp + Ala
-
?
Asp-Ala + H2O
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Lactobacillus delbrueckii subsp. lactis DSM 20072 Asp + Ala
-
?
Glu-Ala + H2O
-
Lactobacillus delbrueckii subsp. lactis Glu + Ala
-
?
Glu-Ala + H2O
-
Lactobacillus delbrueckii subsp. lactis DSM 20072 Glu + Ala
-
?
Glu-Gly + H2O
-
Lactobacillus delbrueckii subsp. lactis Glu + Gly
-
?
Glu-Gly + H2O
-
Lactobacillus delbrueckii subsp. lactis DSM 20072 Glu + Gly
-
?
L-aspartate-beta-naphthylamide + H2O
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Lactobacillus delbrueckii subsp. lactis L-aspartate + beta-naphthylamine
-
?
L-glutamyl-beta-naphthylamide + H2O
-
Lactobacillus delbrueckii subsp. lactis L-glutamine + beta-naphthylamine
-
?
additional information the wild-type enzyme has a high specificity for N-terminal Asp and Glu. Enzyme specificities of the enzyme-metal variants with different substrates, overview Lactobacillus delbrueckii subsp. lactis ?
-
?
additional information the wild-type enzyme has a high specificity for N-terminal Asp and Glu. Enzyme specificities of the enzyme-metal variants with different substrates, overview Lactobacillus delbrueckii subsp. lactis DSM 20072 ?
-
?
Ser-Ala + H2O
-
Lactobacillus delbrueckii subsp. lactis Ser + Ala
-
?
Ser-Ala + H2O
-
Lactobacillus delbrueckii subsp. lactis DSM 20072 Ser + Ala
-
?

Synonyms

Synonyms Comment Organism
aminopeptidase A
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Lactobacillus delbrueckii subsp. lactis
Lb-PepA
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Lactobacillus delbrueckii subsp. lactis
PepA
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Lactobacillus delbrueckii subsp. lactis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
additional information
-
at 60°C, relative activity of 78% is detected for CoCo (PepA). CuCu(PepA) shows a relative activity of 29% at this temperature Lactobacillus delbrueckii subsp. lactis
55
-
CuCu-PepA Lactobacillus delbrueckii subsp. lactis
60
-
MnMn-PepA, NiNi-PepA, and ZnZn-PepA Lactobacillus delbrueckii subsp. lactis
65
-
CoCo-PepA Lactobacillus delbrueckii subsp. lactis

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 80 activity range, profile overview Lactobacillus delbrueckii subsp. lactis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
MnMn-PepA, NiNi-PepA, CoCo-PepA, and ZnZn-PepA Lactobacillus delbrueckii subsp. lactis
7
-
CuCu-PepA Lactobacillus delbrueckii subsp. lactis

pH Range

pH Minimum pH Maximum Comment Organism
4 8 activity range of CuCu-PepA, profile overview Lactobacillus delbrueckii subsp. lactis
4.5 8 activity range of CoCo-PepA, NiNi-PepA, and ZnZn-PepA, profiles overview Lactobacillus delbrueckii subsp. lactis
5 8 activity range of MnMn-PepA, profile overview Lactobacillus delbrueckii subsp. lactis

General Information

General Information Comment Organism
physiological function aminopeptidase A (PepA) belongs to the group of metallopeptidases with two bound metal ions per subunit, M1M2-PepA, it is specific for the cleavage of N-terminal glutamic (Glu) and aspartic acid (Asp) and, in low amounts, serine (Ser) residues Lactobacillus delbrueckii subsp. lactis