BRENDA - Enzyme Database
show all sequences of 3.4.11.25

Mutation of active site serine residue with cysteine displays change in acyl-acceptor preference of beta-peptidyl aminopeptidase from Pseudomonas aeruginosa PAO1

Arima, J.; Tanaka, A.; Morimoto, M.; Mori, N.; Appl. Microbiol. Biotechnol. 98, 1631-1640 (2014)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
-
Pseudomonas aeruginosa
Engineering
Protein Variants
Commentary
Organism
S237C
mutant synthesizes beta-homoGly-beta-homoGly-4-nitroanilide from beta-homoGly-4-nitroanilide efficiently without degradation, indicating that because of the mutation, the enzyme came to recognize only the amine group as an acyl acceptor instead of water. When using cysteamine as an acyl acceptor, beta-homoGly-cysteamine is synthesized by the mutant only in the reaction using S237C. Mutant S237C is unable to synthesize beta-homoGly-cystamine when using cystamine as an acyl acceptor, although it is synthesized by wild-type
Pseudomonas aeruginosa
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0073
-
beta-homoGly 4-nitroanilide
wild-type, pH 9.0, 25°C
Pseudomonas aeruginosa
0.56
-
beta-homoGly 4-nitroanilide
wild-type, pH 6.0, 25°C
Pseudomonas aeruginosa
0.72
-
beta-homoGly 4-nitroanilide
mutant S237C, pH 9.0, 25°C
Pseudomonas aeruginosa
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
14000
-
x * 25000, and x * 14000, SDS-PAGE
Pseudomonas aeruginosa
25000
-
x * 25000, and x * 14000, SDS-PAGE
Pseudomonas aeruginosa
42000
-
gel filtration
Pseudomonas aeruginosa
Organism
Organism
UniProt
Commentary
Textmining
Pseudomonas aeruginosa
Q9I3M6
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2 beta-homoGly 4-nitroanilide
-
731203
Pseudomonas aeruginosa
beta-homoGly-beta-homoGly 4-nitroanilide + 4-nitroaniline
product of mutant S237C, synthesized efficiently without degradation
-
-
?
beta-homoGly 4-nitroanilide + H2O
-
731203
Pseudomonas aeruginosa
beta-homoGly + 4-nitroaniline
wild-type enzyme shows beta-homoGly-beta-homoGly-4-nitroanilide synthetic activity, but the product vanishes in a few min and converts to free beta-homoGly
-
-
?
Subunits
Subunits
Commentary
Organism
dimer
x * 25000, and x * 14000, SDS-PAGE
Pseudomonas aeruginosa
Synonyms
Synonyms
Commentary
Organism
BapF
-
Pseudomonas aeruginosa
PA1486
-
Pseudomonas aeruginosa
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
50
-
wild-type, stable up to
Pseudomonas aeruginosa
60
-
mutant S237C, stable up to
Pseudomonas aeruginosa
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1090
-
beta-homoGly 4-nitroanilide
wild-type, pH 9.0, 25°C
Pseudomonas aeruginosa
1410
-
beta-homoGly 4-nitroanilide
wild-type, pH 6.0, 25°C
Pseudomonas aeruginosa
4540
-
beta-homoGly 4-nitroanilide
mutant S237C, pH 9.0, 25°C
Pseudomonas aeruginosa
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6
6.5
wild-type
Pseudomonas aeruginosa
8
9.5
mutant S237C
Pseudomonas aeruginosa
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
6
10
-
Pseudomonas aeruginosa
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Pseudomonas aeruginosa
Engineering (protein specific)
Protein Variants
Commentary
Organism
S237C
mutant synthesizes beta-homoGly-beta-homoGly-4-nitroanilide from beta-homoGly-4-nitroanilide efficiently without degradation, indicating that because of the mutation, the enzyme came to recognize only the amine group as an acyl acceptor instead of water. When using cysteamine as an acyl acceptor, beta-homoGly-cysteamine is synthesized by the mutant only in the reaction using S237C. Mutant S237C is unable to synthesize beta-homoGly-cystamine when using cystamine as an acyl acceptor, although it is synthesized by wild-type
Pseudomonas aeruginosa
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0073
-
beta-homoGly 4-nitroanilide
wild-type, pH 9.0, 25°C
Pseudomonas aeruginosa
0.56
-
beta-homoGly 4-nitroanilide
wild-type, pH 6.0, 25°C
Pseudomonas aeruginosa
0.72
-
beta-homoGly 4-nitroanilide
mutant S237C, pH 9.0, 25°C
Pseudomonas aeruginosa
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
14000
-
x * 25000, and x * 14000, SDS-PAGE
Pseudomonas aeruginosa
25000
-
x * 25000, and x * 14000, SDS-PAGE
Pseudomonas aeruginosa
42000
-
gel filtration
Pseudomonas aeruginosa
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2 beta-homoGly 4-nitroanilide
-
731203
Pseudomonas aeruginosa
beta-homoGly-beta-homoGly 4-nitroanilide + 4-nitroaniline
product of mutant S237C, synthesized efficiently without degradation
-
-
?
beta-homoGly 4-nitroanilide + H2O
-
731203
Pseudomonas aeruginosa
beta-homoGly + 4-nitroaniline
wild-type enzyme shows beta-homoGly-beta-homoGly-4-nitroanilide synthetic activity, but the product vanishes in a few min and converts to free beta-homoGly
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
x * 25000, and x * 14000, SDS-PAGE
Pseudomonas aeruginosa
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
50
-
wild-type, stable up to
Pseudomonas aeruginosa
60
-
mutant S237C, stable up to
Pseudomonas aeruginosa
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1090
-
beta-homoGly 4-nitroanilide
wild-type, pH 9.0, 25°C
Pseudomonas aeruginosa
1410
-
beta-homoGly 4-nitroanilide
wild-type, pH 6.0, 25°C
Pseudomonas aeruginosa
4540
-
beta-homoGly 4-nitroanilide
mutant S237C, pH 9.0, 25°C
Pseudomonas aeruginosa
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6
6.5
wild-type
Pseudomonas aeruginosa
8
9.5
mutant S237C
Pseudomonas aeruginosa
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
6
10
-
Pseudomonas aeruginosa
Other publictions for EC 3.4.11.25
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
753761
Kolesinska
-
Labeling and protecting N-ter ...
Sphingosinicella xenopeptidilytica
Helv. Chim. Acta
101
e1700259
2018
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
731203
Arima
Mutation of active site serine ...
Pseudomonas aeruginosa
Appl. Microbiol. Biotechnol.
98
1631-1640
2014
-
-
1
-
1
-
-
3
-
-
3
-
-
1
-
-
-
-
-
-
-
-
2
1
2
-
-
2
3
2
-
1
-
-
-
-
-
-
1
-
-
1
-
-
-
-
3
-
-
3
-
-
-
-
-
-
-
-
-
2
1
-
-
2
3
2
-
1
-
-
-
-
-
-
-
708080
Heck
Enzyme-catalyzed formation of ...
Sphingosinicella microcystinivorans, Sphingosinicella microcystinivorans Y2, Sphingosinicella xenopeptidilytica, Sphingosinicella xenopeptidilytica 3-2W4
Chem. Biodivers.
4
2016-2030
2007
-
2
-
-
-
-
-
-
-
-
-
-
-
13
-
-
-
-
-
-
-
-
4
-
2
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
708078
Heck
Enzymatic degradation of beta- ...
Sphingosinicella microcystinivorans, Sphingosinicella microcystinivorans Y2, Sphingosinicella xenopeptidilytica, Sphingosinicella xenopeptidilytica 3-2W4
Chem. Biodivers.
3
1325-1348
2006
-
-
-
-
-
-
-
6
-
-
-
-
-
9
-
-
2
-
-
-
-
-
60
-
2
2
-
-
6
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
2
-
-
-
-
60
-
2
-
-
6
2
-
-
-
-
-
-
-
6
6
708467
Geueke
Bacterial beta-peptidyl aminop ...
Sphingosinicella microcystinivorans, Sphingosinicella microcystinivorans Y2, Sphingosinicella xenopeptidilytica, Sphingosinicella xenopeptidilytica 3-2W4
FEBS J.
273
5261-5272
2006
-
-
-
-
-
-
4
6
-
-
8
-
-
9
-
1
2
-
-
-
2
-
46
1
2
2
-
4
6
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
4
-
6
-
-
8
-
-
-
1
2
-
-
2
-
46
1
2
-
4
6
4
-
-
-
-
-
-
-
8
8
708917
Geueke
A novel beta-peptidyl aminopep ...
Sphingosinicella xenopeptidilytica, Sphingosinicella xenopeptidilytica 3-2W4
J. Bacteriol.
187
5910-5917
2005
1
-
-
-
-
-
1
-
-
1
2
-
-
8
-
1
1
-
-
-
1
-
4
1
-
1
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
-
-
-
1
2
-
-
-
1
1
-
-
1
-
4
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-