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Literature summary for 1.2.1.16 extracted from
- Kinetic and structural insights into enzymatic mechanism of succinic semialdehyde dehydrogenase from Cyanothece sp. ATCC51142 (2020), PLoS ONE, 15, e0239372 .
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | - |
Crocosphaera subtropica |  |
| DTT | - |
Crocosphaera subtropica | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene gabD or cce4228, DNA and amino acid sequence deternination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Crocosphaera subtropica |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K154A | site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type enzyme | Crocosphaera subtropica |
| S157E | site-directed mutagenesis, the mutant shows reduced activity with NAD+/SSA and increased activity with NADP+/SSA compared to wild-type enzyme | Crocosphaera subtropica |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | causes a 2fold decrease in activity at 2 mM | Crocosphaera subtropica | |
| succinate semialdehyde | competitive inhibition by the substrate | Crocosphaera subtropica | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics, steady-state kinetics of cce4228 wild-type and mutants enzymes | Crocosphaera subtropica | |
| 0.0028 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NAD+ | Crocosphaera subtropica | |
| 0.003 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NADP+ | Crocosphaera subtropica | |
| 0.004 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NAD+ | Crocosphaera subtropica | |
| 0.008 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NADP+ | Crocosphaera subtropica | |
| 0.02 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NAD+ | Crocosphaera subtropica | |
| 0.02 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NADP+ | Crocosphaera subtropica | |
| 0.034 | - |
NADP+ | pH 9.0, 25°C, recombinant wild-type enzyme | Crocosphaera subtropica | |
| 0.51 | - |
NAD+ | pH 9.0, 25°C, recombinant mutant S157E | Crocosphaera subtropica | |
| 1.1 | - |
NADP+ | pH 9.0, 25°C, recombinant mutant S157E | Crocosphaera subtropica | |
| 1.5 | - |
NAD+ | pH 9.0, 25°C, recombinant mutant K154A | Crocosphaera subtropica | |
| 1.7 | - |
NAD+ | pH 9.0, 25°C, recombinant wild-type enzyme | Crocosphaera subtropica | |
| 6.1 | - |
NADP+ | pH 9.0, 25°C, recombinant mutant v | Crocosphaera subtropica | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required, Km = 0.00624 mM | Crocosphaera subtropica | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 109000 | - |
recombinant His6-tagged enzyme, gel filtration | Crocosphaera subtropica |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| succinate semialdehyde + NAD+ + H2O | Crocosphaera subtropica | - |
succinate + NADH + 2 H+ | - |
? | |
| succinate semialdehyde + NAD+ + H2O | Crocosphaera subtropica BH68 | - |
succinate + NADH + 2 H+ | - |
? | |
| succinate semialdehyde + NADP+ + H2O | Crocosphaera subtropica | - |
succinate + NADPH + 2 H+ | - |
? | |
| succinate semialdehyde + NADP+ + H2O | Crocosphaera subtropica BH68 | - |
succinate + NADPH + 2 H+ | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Crocosphaera subtropica | B1WSJ7 | Cyanothece sp. ATCC 51142 | - |
| Crocosphaera subtropica BH68 | B1WSJ7 | Cyanothece sp. ATCC 51142 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Crocosphaera subtropica |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| succinate semialdehyde + NAD(P)+ + H2O = succinate + NAD(P)H + H+ | proposed enzymatic mechanism of cce4228 protein, overview | Crocosphaera subtropica |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | protein cce4228 shows similar catalytic efficiency towards succinate semialdehyde (SSA) at saturated concentration of NAD+ or NADP+ | Crocosphaera subtropica | ? | - |
- |
|
| additional information | protein cce4228 shows similar catalytic efficiency towards succinate semialdehyde (SSA) at saturated concentration of NAD+ or NADP+ | Crocosphaera subtropica BH68 | ? | - |
- |
|
| succinate semialdehyde + NAD+ + H2O | - |
Crocosphaera subtropica | succinate + NADH + 2 H+ | - |
? | |
| succinate semialdehyde + NAD+ + H2O | - |
Crocosphaera subtropica BH68 | succinate + NADH + 2 H+ | - |
? | |
| succinate semialdehyde + NADP+ + H2O | - |
Crocosphaera subtropica | succinate + NADPH + 2 H+ | - |
? | |
| succinate semialdehyde + NADP+ + H2O | - |
Crocosphaera subtropica BH68 | succinate + NADPH + 2 H+ | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 50000, recombinant His6-tagged enzyme, SDS-PAGE, 2 * 52518, His6-tagged enzyme, sequence calculation | Crocosphaera subtropica |
| More | the Cys262 in the active site of cce4228 does not form a disulfide bond with other Cys residues, although there are six Cys residues in cce4228 protein | Crocosphaera subtropica |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| cce4228 | - |
Crocosphaera subtropica |
| cce4228 protein | - |
Crocosphaera subtropica |
| gabD | - |
Crocosphaera subtropica |
| SSADH | - |
Crocosphaera subtropica |
| succinic semialdehyde dehydrogenase | - |
Crocosphaera subtropica |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Crocosphaera subtropica |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.1 | - |
NADP+ | pH 9.0, 25°C, recombinant mutant K154A | Crocosphaera subtropica | |
| 0.1 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NADP+ | Crocosphaera subtropica | |
| 0.13 | - |
NAD+ | pH 9.0, 25°C, recombinant mutant K154A | Crocosphaera subtropica | |
| 0.13 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NAD+ | Crocosphaera subtropica | |
| 0.58 | - |
NAD+ | pH 9.0, 25°C, recombinant mutant S157E | Crocosphaera subtropica | |
| 0.58 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NAD+ | Crocosphaera subtropica | |
| 1.6 | - |
NADP+ | pH 9.0, 25°C, recombinant mutant S157E | Crocosphaera subtropica | |
| 1.6 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NADP+ | Crocosphaera subtropica | |
| 1.74 | - |
NAD+ | pH 9.0, 25°C, recombinant wild-type enzyme | Crocosphaera subtropica | |
| 1.74 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NAD+ | Crocosphaera subtropica | |
| 3.9 | - |
NADP+ | pH 9.0, 25°C, recombinant wild-type enzyme | Crocosphaera subtropica | |
| 3.9 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NADP+ | Crocosphaera subtropica | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 9 | - |
assay at | Crocosphaera subtropica |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | although the binding affinity of cce4228 protein with NAD+ is about 50fold lower than that of cce4228 with NADP+, the catalytic efficiency of cce4228 protein towards succinic semialdehyde with saturated concentration of NADP+ is same as that with saturated concentration of NAD+ as its cofactors. Protein cce4228 harbors Ser157 and Lys154 residues in its active center which might determine the cofactor preference | Crocosphaera subtropica | |
| NAD+ | - |
Crocosphaera subtropica | |
| NADP+ | - |
Crocosphaera subtropica | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.02 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NAD+ | Crocosphaera subtropica | |
| 0.02 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NADP+ | Crocosphaera subtropica | |
| 0.12 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NADP+ | Crocosphaera subtropica | |
| 0.13 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NAD+ | Crocosphaera subtropica | |
| 0.52 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NAD+ | Crocosphaera subtropica | |
| 0.8 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NADP+ | Crocosphaera subtropica | |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | the conserved Cys262 and Glu228 residues are crucial for the catalytic activity of cce4228 protein and the Ser157 and Lys154 residues are determinants of cofactor preference. The Cys residue is thought to function as a nucleophile in the catalytic mechanism. Homology structure modeling of wild-type cce4228 protein | Crocosphaera subtropica |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.016 | - |
NADP+ | pH 9.0, 25°C, recombinant mutant v | Crocosphaera subtropica | |
| 0.087 | - |
NAD+ | pH 9.0, 25°C, recombinant mutant K154A | Crocosphaera subtropica | |
| 1.02 | - |
NAD+ | pH 9.0, 25°C, recombinant wild-type enzyme | Crocosphaera subtropica | |
| 1.14 | - |
NAD+ | pH 9.0, 25°C, recombinant mutant S157E | Crocosphaera subtropica | |
| 1.46 | - |
NADP+ | pH 9.0, 25°C, recombinant mutant S157E | Crocosphaera subtropica | |
| 5 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NADP+ | Crocosphaera subtropica | |
| 6.5 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant K154A, with NAD+ | Crocosphaera subtropica | |
| 114.7 | - |
NADP+ | pH 9.0, 25°C, recombinant wild-type enzyme | Crocosphaera subtropica | |
| 207.1 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NAD+ | Crocosphaera subtropica | |
| 435 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NAD+ | Crocosphaera subtropica | |
| 487.5 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant wild-type enzyme, with NADP+ | Crocosphaera subtropica | |
| 533.3 | - |
succinate semialdehyde | pH 9.0, 25°C, recombinant mutant S157E, with NADP+ | Crocosphaera subtropica | |
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