Activating Compound | Comment | 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 | 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 | Comment | 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 Maximum [mM] | Substrate | Comment | 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 | Comment | 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 Maximum [Da] | Comment | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Vibrio alginolyticus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
to homogeneity, yield 33%, 1800fold purified, ammonium sulfate precipitation, DEAE-Sepharose CL-6B, hydroxyapatite, and Blue Sepharose CL-6B chromatography | Vibrio alginolyticus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | 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 | 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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
carboxynorspermidine + NAD+ + H2O | NADH is a much poorer cofactor than NADPH | Vibrio alginolyticus | L-aspartic 4-semialdehyde + 1,3-diaminopropane + NADH + H+ | - |
ir | |
carboxynorspermidine + NAD+ + H2O | NADH is a much poorer cofactor than NADPH | 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 | 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 | 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 | 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 | Vibrio alginolyticus ATCC 17749 | L-aspartic 4-semialdehyde + putrescine + NADPH + H+ | - |
ir | |
additional information | no activity with ethylenediamine, cadaverine, and D-aspartic beta-semialdehyde | Vibrio alginolyticus | ? | - |
? | |
additional information | no activity with ethylenediamine, cadaverine, and D-aspartic beta-semialdehyde | Vibrio alginolyticus ATCC 17749 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 45100, SDS-PAGE, indicates that the native enzyme is composed of two subunits of similar size | Vibrio alginolyticus |
Synonyms | Comment | Organism |
---|---|---|
C-NSPD synthase | - |
Vibrio alginolyticus |
carboxynorspermidine synthase | - |
Vibrio alginolyticus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
- |
Vibrio alginolyticus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.25 | 7.5 | - |
Vibrio alginolyticus |
pH Minimum | pH Maximum | Comment | 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 Maximum | Comment | 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 | Comment | 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 |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Vibrio alginolyticus | isoelectric focusing and chromatofocusing | 4.3 | 4.2 |
General Information | Comment | 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 |