BRENDA - Enzyme Database show
show all sequences of 1.4.1.B4

Purification and some properties of carboxynorspermidine synthase participating in a novel biosynthetic pathway for norspermidine inVibrio alginolyticus

Nakao, H.; Shinoda, S.; Yamamoto, S.; J. Gen. Microbiol. 137, 1737-1742 (1991)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
DTT
without addition 7% of the maximum activity detected in the purified enzyme, 14fold activiation is observed at 20 mM
Vibrio alginolyticus
additional information
1 mM EDTA shows no stimulatory effect on enzyme activity
Vibrio alginolyticus
General Stability
General Stability
Organism
no significant stabilization by addition of NADPH. 1 mM DTT has a protective effect on the enzyme during purification and storage. Freezing and thawing results in almost complete loss of activity
Vibrio alginolyticus
Inhibitors
Inhibitors
Commentary
Organism
Structure
1,2-diaminopropane
10 mM, causes 10-20% inhibition of carboxynorspermidine formation
Vibrio alginolyticus
cadaverine
10 mM, causes 10-20% inhibition of carboxynorspermidine formation
Vibrio alginolyticus
ethylenediamine
10 mM, causes 10-20% inhibition of carboxynorspermidine formation
Vibrio alginolyticus
ethylmaleimide
5 mM, in presence of 20 mM, 83% inhibition
Vibrio alginolyticus
iodoacetamide
5 mM, in presence of 20 mM, 24% inhibition
Vibrio alginolyticus
additional information
no inhibitory effect of NAD+, ATP, and spermidine at 5 or 10 mM. No effect of 1 mM Na+ or K+. No evidence for inhibitory effect of metal cations such as Ca2+, Mg2+, Fe3+, Fe2+, Cu+, Cu2+, Mn2+, and Zn2+. EDTA, 1 mM no inhibition
Vibrio alginolyticus
NADP+
5 mM, 51% inhibition
Vibrio alginolyticus
norspermidine
10 mM, 10% inhibition
Vibrio alginolyticus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
the Km for the Schiff base is 4.68 mM, the intermediate formed from L-aspartic 4-semialdehyde and 1,3-diaminopropane, which is reduced to carboxynorspermidine
Vibrio alginolyticus
1.51
-
NADPH
pH 7.5, 37°C, with a Vmax of 31 micromol carboxynorspermidine/min/mg protein formed
Vibrio alginolyticus
2.97
-
NADH
pH 7.5, 37°C, with a Vmax of 13.5 micromol carboxynorspermidine/min/mg protein formed
Vibrio alginolyticus
4.68
-
L-aspartic 4-semialdehyde
pH 7.5, 37°C, Vmax: 35 micromol/min/mg carboxynorspermidine
Vibrio alginolyticus
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
no evidence for activity being dependent on metal ions such as Ca2+, Mg2+, Fe3+, Fe2+, Cu+, Cu2+, Mn2+, and Zn2+
Vibrio alginolyticus
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
45100
-
2 * 45100, SDS-PAGE, indicates that the native enzyme is composed of two subunits of similar size
Vibrio alginolyticus
93500
-
gel filtration
Vibrio alginolyticus
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Vibrio alginolyticus
-
-
-
Purification (Commentary)
Commentary
Organism
to homogeneity, yield 33%, 1800fold purified, ammonium sulfate precipitation, DEAE-Sepharose CL-6B, hydroxyapatite, and Blue Sepharose CL-6B chromatography
Vibrio alginolyticus
Reaction
Reaction
Commentary
Organism
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADPH + H+ = carboxynorspermidine + NADP+ + H2O
i.e. (3-((3-aminopropyl)amino)propyl)carbamate (carboxynorspermidine) and (2S)-2-amino-4-oxobutanoate (L-aspartic 4-semialdehyde) via a nicotinamide-nucleotide reduction of the Schiff base H2N(CH2)3=CHCH2CH(NH2)COOH, formed from L-aspartic beta-semialdehyde, an intermediate in the novel pathway for norspermidine biosynthesis
Vibrio alginolyticus
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
in the presence of 5 mM nospermidine, specific activity is reduced by 70%
Vibrio alginolyticus
0.0039
-
addition of 5 mM norspermidine to the growth medium, 27% compared to the specific activity without addition, pH 7.5, 37°C
Vibrio alginolyticus
0.0108
-
addition of 5 mM spermidine to the growth medium, 74% compared to the specific activity without addition, pH 7.5, 37°C
Vibrio alginolyticus
0.01455
-
without addition of norspermidine or spermidine to the growth medium, pH 7.5, 37°C
Vibrio alginolyticus
31
-
micromol carboxynorspermidine/min-1/mg protein, pH 7.5, 37°C
Vibrio alginolyticus
Storage Stability
Storage Stability
Organism
4°C, protein concentration of 1.6 mg/ml in 20 mM Tris-HCl, pH 7.5, 1 mM DTT and 0.02% NaN3, lost of about 15% of its activity after one week
Vibrio alginolyticus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
carboxynorspermidine + NAD+ + H2O
NADH is a much poorer cofactor than NADPH
712623
Vibrio alginolyticus
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADH + H+
-
-
-
ir
carboxynorspermidine + NAD+ + H2O
NADH is a much poorer cofactor than NADPH
712623
Vibrio alginolyticus ATCC 17749
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADH + H+
-
-
-
ir
carboxynorspermidine + NADP+ + H2O
i.e. (3-((3-aminopropyl)amino)propyl)carbamate, incubation of the purified enzyme in the range of pH 6-8, with cofactor NADP+ or NAD+ concentrations up to 5 mM and substrate concentrations of carboxynorspermidine up to 10 mM, no reversible activity is observed
712623
Vibrio alginolyticus
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADPH + H+
i.e. (2S)-2-amino-4-oxobutanoate
-
-
ir
carboxynorspermidine + NADP+ + H2O
i.e. (3-((3-aminopropyl)amino)propyl)carbamate, incubation of the purified enzyme in the range of pH 6-8, with cofactor NADP+ or NAD+ concentrations up to 5 mM and substrate concentrations of carboxynorspermidine up to 10 mM, no reversible activity is observed
712623
Vibrio alginolyticus ATCC 17749
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADPH + H+
i.e. (2S)-2-amino-4-oxobutanoate
-
-
ir
carboxyspermidine + NADP+ + H2O
i.e. 3-[(4-aminobutyl)amino]-L-alanine, slightly active with putrescine, ca. 7% of the rate compared to 1,3-diaminopropane
712623
Vibrio alginolyticus
L-aspartic 4-semialdehyde + putrescine + NADPH + H+
-
-
-
ir
carboxyspermidine + NADP+ + H2O
i.e. 3-[(4-aminobutyl)amino]-L-alanine, slightly active with putrescine, ca. 7% of the rate compared to 1,3-diaminopropane
712623
Vibrio alginolyticus ATCC 17749
L-aspartic 4-semialdehyde + putrescine + NADPH + H+
-
-
-
ir
additional information
no activity with ethylenediamine, cadaverine, and D-aspartic beta-semialdehyde
712623
Vibrio alginolyticus
?
-
-
-
-
additional information
no activity with ethylenediamine, cadaverine, and D-aspartic beta-semialdehyde
712623
Vibrio alginolyticus ATCC 17749
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
dimer
2 * 45100, SDS-PAGE, indicates that the native enzyme is composed of two subunits of similar size
Vibrio alginolyticus
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
-
Vibrio alginolyticus
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
45
78 and 59% of the maximal activity at 30 and 45°C, respectively, no activity at 50°C
Vibrio alginolyticus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.25
7.5
-
Vibrio alginolyticus
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6.25
8
18 and 85% of the maximal acticity at pH 6.25 and 8.0, respectively
Vibrio alginolyticus
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
6.5
-
the enzyme is unstable in buffers below pH 6.5, at pH 6.5 50% loss of activity is observed within 12 h at 4°C
Vibrio alginolyticus
Cofactor
Cofactor
Commentary
Organism
Structure
NADH
conmpletely inactive unless NADPH or NADH are added, NADH is a much poorer cofactor than NADPH
Vibrio alginolyticus
NADPH
conmpletely inactive unless NADPH or NADH are added, NADH is a much poorer cofactor than NADPH
Vibrio alginolyticus
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Vibrio alginolyticus
isoelectric focusing and chromatofocusing
4.3
4.2
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
DTT
without addition 7% of the maximum activity detected in the purified enzyme, 14fold activiation is observed at 20 mM
Vibrio alginolyticus
additional information
1 mM EDTA shows no stimulatory effect on enzyme activity
Vibrio alginolyticus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADH
conmpletely inactive unless NADPH or NADH are added, NADH is a much poorer cofactor than NADPH
Vibrio alginolyticus
NADPH
conmpletely inactive unless NADPH or NADH are added, NADH is a much poorer cofactor than NADPH
Vibrio alginolyticus
General Stability (protein specific)
General Stability
Organism
no significant stabilization by addition of NADPH. 1 mM DTT has a protective effect on the enzyme during purification and storage. Freezing and thawing results in almost complete loss of activity
Vibrio alginolyticus
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
1,2-diaminopropane
10 mM, causes 10-20% inhibition of carboxynorspermidine formation
Vibrio alginolyticus
cadaverine
10 mM, causes 10-20% inhibition of carboxynorspermidine formation
Vibrio alginolyticus
ethylenediamine
10 mM, causes 10-20% inhibition of carboxynorspermidine formation
Vibrio alginolyticus
ethylmaleimide
5 mM, in presence of 20 mM, 83% inhibition
Vibrio alginolyticus
iodoacetamide
5 mM, in presence of 20 mM, 24% inhibition
Vibrio alginolyticus
additional information
no inhibitory effect of NAD+, ATP, and spermidine at 5 or 10 mM. No effect of 1 mM Na+ or K+. No evidence for inhibitory effect of metal cations such as Ca2+, Mg2+, Fe3+, Fe2+, Cu+, Cu2+, Mn2+, and Zn2+. EDTA, 1 mM no inhibition
Vibrio alginolyticus
NADP+
5 mM, 51% inhibition
Vibrio alginolyticus
norspermidine
10 mM, 10% inhibition
Vibrio alginolyticus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
the Km for the Schiff base is 4.68 mM, the intermediate formed from L-aspartic 4-semialdehyde and 1,3-diaminopropane, which is reduced to carboxynorspermidine
Vibrio alginolyticus
1.51
-
NADPH
pH 7.5, 37°C, with a Vmax of 31 micromol carboxynorspermidine/min/mg protein formed
Vibrio alginolyticus
2.97
-
NADH
pH 7.5, 37°C, with a Vmax of 13.5 micromol carboxynorspermidine/min/mg protein formed
Vibrio alginolyticus
4.68
-
L-aspartic 4-semialdehyde
pH 7.5, 37°C, Vmax: 35 micromol/min/mg carboxynorspermidine
Vibrio alginolyticus
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
no evidence for activity being dependent on metal ions such as Ca2+, Mg2+, Fe3+, Fe2+, Cu+, Cu2+, Mn2+, and Zn2+
Vibrio alginolyticus
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
45100
-
2 * 45100, SDS-PAGE, indicates that the native enzyme is composed of two subunits of similar size
Vibrio alginolyticus
93500
-
gel filtration
Vibrio alginolyticus
Purification (Commentary) (protein specific)
Commentary
Organism
to homogeneity, yield 33%, 1800fold purified, ammonium sulfate precipitation, DEAE-Sepharose CL-6B, hydroxyapatite, and Blue Sepharose CL-6B chromatography
Vibrio alginolyticus
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
in the presence of 5 mM nospermidine, specific activity is reduced by 70%
Vibrio alginolyticus
0.0039
-
addition of 5 mM norspermidine to the growth medium, 27% compared to the specific activity without addition, pH 7.5, 37°C
Vibrio alginolyticus
0.0108
-
addition of 5 mM spermidine to the growth medium, 74% compared to the specific activity without addition, pH 7.5, 37°C
Vibrio alginolyticus
0.01455
-
without addition of norspermidine or spermidine to the growth medium, pH 7.5, 37°C
Vibrio alginolyticus
31
-
micromol carboxynorspermidine/min-1/mg protein, pH 7.5, 37°C
Vibrio alginolyticus
Storage Stability (protein specific)
Storage Stability
Organism
4°C, protein concentration of 1.6 mg/ml in 20 mM Tris-HCl, pH 7.5, 1 mM DTT and 0.02% NaN3, lost of about 15% of its activity after one week
Vibrio alginolyticus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
carboxynorspermidine + NAD+ + H2O
NADH is a much poorer cofactor than NADPH
712623
Vibrio alginolyticus
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADH + H+
-
-
-
ir
carboxynorspermidine + NAD+ + H2O
NADH is a much poorer cofactor than NADPH
712623
Vibrio alginolyticus ATCC 17749
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADH + H+
-
-
-
ir
carboxynorspermidine + NADP+ + H2O
i.e. (3-((3-aminopropyl)amino)propyl)carbamate, incubation of the purified enzyme in the range of pH 6-8, with cofactor NADP+ or NAD+ concentrations up to 5 mM and substrate concentrations of carboxynorspermidine up to 10 mM, no reversible activity is observed
712623
Vibrio alginolyticus
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADPH + H+
i.e. (2S)-2-amino-4-oxobutanoate
-
-
ir
carboxynorspermidine + NADP+ + H2O
i.e. (3-((3-aminopropyl)amino)propyl)carbamate, incubation of the purified enzyme in the range of pH 6-8, with cofactor NADP+ or NAD+ concentrations up to 5 mM and substrate concentrations of carboxynorspermidine up to 10 mM, no reversible activity is observed
712623
Vibrio alginolyticus ATCC 17749
L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADPH + H+
i.e. (2S)-2-amino-4-oxobutanoate
-
-
ir
carboxyspermidine + NADP+ + H2O
i.e. 3-[(4-aminobutyl)amino]-L-alanine, slightly active with putrescine, ca. 7% of the rate compared to 1,3-diaminopropane
712623
Vibrio alginolyticus
L-aspartic 4-semialdehyde + putrescine + NADPH + H+
-
-
-
ir
carboxyspermidine + NADP+ + H2O
i.e. 3-[(4-aminobutyl)amino]-L-alanine, slightly active with putrescine, ca. 7% of the rate compared to 1,3-diaminopropane
712623
Vibrio alginolyticus ATCC 17749
L-aspartic 4-semialdehyde + putrescine + NADPH + H+
-
-
-
ir
additional information
no activity with ethylenediamine, cadaverine, and D-aspartic beta-semialdehyde
712623
Vibrio alginolyticus
?
-
-
-
-
additional information
no activity with ethylenediamine, cadaverine, and D-aspartic beta-semialdehyde
712623
Vibrio alginolyticus ATCC 17749
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
2 * 45100, SDS-PAGE, indicates that the native enzyme is composed of two subunits of similar size
Vibrio alginolyticus
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
-
Vibrio alginolyticus
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
45
78 and 59% of the maximal activity at 30 and 45°C, respectively, no activity at 50°C
Vibrio alginolyticus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.25
7.5
-
Vibrio alginolyticus
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6.25
8
18 and 85% of the maximal acticity at pH 6.25 and 8.0, respectively
Vibrio alginolyticus
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
6.5
-
the enzyme is unstable in buffers below pH 6.5, at pH 6.5 50% loss of activity is observed within 12 h at 4°C
Vibrio alginolyticus
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Vibrio alginolyticus
isoelectric focusing and chromatofocusing
4.3
4.2
General Information
General Information
Commentary
Organism
physiological function
carboxynorspermidine synthase participates in the novel pathway for norspermidine biosynthesis, with two other enzymes, 2,4-diamonobutyrate deacroxylase and carboxynorspermidine decarboxylase
Vibrio alginolyticus
General Information (protein specific)
General Information
Commentary
Organism
physiological function
carboxynorspermidine synthase participates in the novel pathway for norspermidine biosynthesis, with two other enzymes, 2,4-diamonobutyrate deacroxylase and carboxynorspermidine decarboxylase
Vibrio alginolyticus
Other publictions for EC 1.4.1.B4
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
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)
719993
Hanfrey
Alternative spermidine biosynt ...
Campylobacter jejuni
J. Biol. Chem.
286
43301-43312
2011
-
-
-
-
1
-
-
-
-
-
-
1
-
7
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
712623
Nakao
Purification and some properti ...
Vibrio alginolyticus
J. Gen. Microbiol.
137
1737-1742
1991
2
-
-
-
-
1
8
4
-
1
2
-
-
1
-
-
1
1
-
-
5
1
8
1
1
1
-
-
1
1
1
2
-
1
-
2
-
-
2
-
-
1
-
8
-
4
-
1
2
-
-
-
-
1
-
-
5
1
8
1
1
1
-
-
1
1
1
1
-
1
1
-
-
-