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Literature summary for 1.2.1.79 extracted from

  • Wang, X.; Lai, C.; Lei, G.; Wang, F.; Long, H.; Wu, X.; Chen, J.; Huo, G.; Li, Z.
    Kinetic characterization and structural modeling of an NADP+-dependent succinic semialdehyde dehydrogenase from Anabaena sp. PCC7120 (2018), Int. J. Biol. Macromol., 108, 615-624 .
    View publication on PubMed
Search for more literature for 1.2.1.79

Cloned(Commentary)

Cloned (Comment) Organism
gene all3556, recombinant expression of wild-type and mutant His-tagged enzymes in Escherichia coli strain BL21(DE3) Nostoc sp. PCC 7120 = FACHB-418

Protein Variants

Protein Variants Comment Organism
E228A site-directed mutagenesis, inactive mutant Nostoc sp. PCC 7120 = FACHB-418
E228D sitedirected mutagenesis, the mutant shows highly reduced activity compared to wild-type enzyme Nostoc sp. PCC 7120 = FACHB-418
N131A site-directed mutagenesis, inactive mutant Nostoc sp. PCC 7120 = FACHB-418
N131D site-directed mutagenesis, inactive mutant Nostoc sp. PCC 7120 = FACHB-418
R139A site-directed mutagenesis, the mutant displays catalytic efficiency (kcat/Km) of only respective 0.2% compared to wild-type enzyme with significantly decreased binding affinity for succinic semialdehyde Nostoc sp. PCC 7120 = FACHB-418
S157E site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type, preferring NAD+, mutation of Ser157 does not significantly affect the binding affinity of SSA with the enzyme Nostoc sp. PCC 7120 = FACHB-418
S157P site-directed mutagenesis, the mutant shows altered cofactor specificity compared to wild-type, preferring NAD+, mutation of Ser157 does not significantly affect the binding affinity of SSA with the enzyme Nostoc sp. PCC 7120 = FACHB-418
S420A site-directed mutagenesis, the mutant displays catalytic efficiency (kcat/Km) of only respective 0.4% compared to wild-type enzyme with significantly decreased binding affinity for succinic semialdehyde Nostoc sp. PCC 7120 = FACHB-418

Inhibitors

Inhibitors Comment Organism Structure
Ca2+
-
 Nostoc sp. PCC 7120 = FACHB-418
Co2+
-
 Nostoc sp. PCC 7120 = FACHB-418
Cu2+
-
 Nostoc sp. PCC 7120 = FACHB-418
EDTA partial inhibition at 2 mM Nostoc sp. PCC 7120 = FACHB-418
Ni2+
-
 Nostoc sp. PCC 7120 = FACHB-418
succinate semialdehyde substrate inhhibition at 2 mM, not at 0.1 mM Nostoc sp. PCC 7120 = FACHB-418
Zn2+
-
 Nostoc sp. PCC 7120 = FACHB-418

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
 NADP+ pH 9.5, 25°C, recombinant wild-type enzyme Nostoc sp. PCC 7120 = FACHB-418
4.49
-
 NAD+ pH 9.5, 25°C, recombinant mutant S157E Nostoc sp. PCC 7120 = FACHB-418
5.98
-
 NAD+ pH 9.5, 25°C, recombinant mutant S157P Nostoc sp. PCC 7120 = FACHB-418
10.4
-
 NADP+ pH 9.5, 25°C, recombinant mutant S157E Nostoc sp. PCC 7120 = FACHB-418
12.9
-
 NADP+ pH 9.5, 25°C, recombinant mutant S157P Nostoc sp. PCC 7120 = FACHB-418
16.1
-
 NAD+ pH 9.5, 25°C, recombinant wild-type enzyme Nostoc sp. PCC 7120 = FACHB-418

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ activates 10-13fold at 2 mM Nostoc sp. PCC 7120 = FACHB-418
Mn2+ activates Nostoc sp. PCC 7120 = FACHB-418

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
85000
-
 recombinant His-tagged enzyme, gel filtration Nostoc sp. PCC 7120 = FACHB-418

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
succinate semialdehyde + NADP+ + H2O Nostoc sp. PCC 7120 = FACHB-418
-
 succinate + NADPH + 2 H+
-
 ?

Organism

Organism UniProt Comment Textmining
Nostoc sp. PCC 7120 = FACHB-418 Q8YR92 Anabaena sp. PCC7120, strain SAG 25.82 / UTEX 2576
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification the N-terminal methionine residue of ApSSADH is removed during posttranslational modification Nostoc sp. PCC 7120 = FACHB-418

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and dialysis, to homogeneity Nostoc sp. PCC 7120 = FACHB-418

Reaction

Reaction Comment Organism Reaction ID
succinate semialdehyde + NADP+ + H2O = succinate + NADPH + 2 H+ the thiol-group of Cys residue in aldehyde dehydrogenase will attack NADP+ to form the adduct is the first step in the catalytic mechanism. The key step is the formation of thiohemiacetal tetrahedral intermediate, which then is converted to thioester intermediate by transferring the hydride to NAD(P)+, catalytic reaction kinetic analysis Nostoc sp. PCC 7120 = FACHB-418
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
succinate semialdehyde + NAD+ + H2O very low activity with wild-type enzyme, higher activity with enzyme mutants S157E and S157P Nostoc sp. PCC 7120 = FACHB-418 succinate + NADH + 2 H+
-
 ?
succinate semialdehyde + NADP+ + H2O
-
 Nostoc sp. PCC 7120 = FACHB-418 succinate + NADPH + 2 H+
-
 ?
succinate semialdehyde + NADP+ + H2O preferred substrates Nostoc sp. PCC 7120 = FACHB-418 succinate + NADPH + 2 H+
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 50000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 50985, His-tagged enzyme, sequence calculation, 2 * 50853, recombinant enzyme, mass spectrometry Nostoc sp. PCC 7120 = FACHB-418

Synonyms

Synonyms Comment Organism
all3556
-
 Nostoc sp. PCC 7120 = FACHB-418
ApSSADH
-
 Nostoc sp. PCC 7120 = FACHB-418
NADP+-dependent SSADH
-
 Nostoc sp. PCC 7120 = FACHB-418
NADP+-dependent succinic semialdehyde dehydrogenase
-
 Nostoc sp. PCC 7120 = FACHB-418
SSADH
-
 Nostoc sp. PCC 7120 = FACHB-418
succinic semialdehyde dehydrogenase
-
 Nostoc sp. PCC 7120 = FACHB-418

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Nostoc sp. PCC 7120 = FACHB-418

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
 recombinant enzyme Nostoc sp. PCC 7120 = FACHB-418

Cofactor

Cofactor Comment Organism Structure
additional information ApSSADH prefers to use NADP+ rather than NAD+ as its cofactor. Residue Ser157 of ApSSADH plays a critical role in determining the cofactor preference. The catalytic activities of mutants S157E and S157P are elevated when the cofactor is switched from NADP+ to NAD+ Nostoc sp. PCC 7120 = FACHB-418
NAD+ preferred cofactor for enzyme mutants S157E and S157P compared to wild-type enzyme Nostoc sp. PCC 7120 = FACHB-418
NADP+ preferred cofactor, the Glu228 residue is located in the NADP+ binding pocket Nostoc sp. PCC 7120 = FACHB-418

General Information

General Information Comment Organism
evolution SSADH belongs to the aldehyde dehydrogenase (ALDH) superfamily, which is a kind of NAD(P)+-dependent oxidoreductase using a wide range of aldehydes as its substrate Nostoc sp. PCC 7120 = FACHB-418
additional information the catalytic active center harbors residues such as Cys262, Glu228, Asn131, Arg139 and Ser420. Structure homology modeling of ApSSADH based on the crystal structure of enzyme SpSSADH (PDB ID 3VZ3) from Synechocystis sp. PCC6803. The residues of Cys262 and Asn131 interact with the carbonyl oxygen atom of SSA through the backbone NH group via hydrogen bond. Meanwhile, the side chains of Ser420 and Arg139 interact with the carboxyl oxygen of SSA directly. In addition, the side chains of Arg139 and Glu228 residues interact with the amide group of NADP+ Nostoc sp. PCC 7120 = FACHB-418

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
52.8
-
 NADP+ pH 9.5, 25°C, recombinant mutant S157E Nostoc sp. PCC 7120 = FACHB-418
126
-
 NADP+ pH 9.5, 25°C, recombinant mutant S157P Nostoc sp. PCC 7120 = FACHB-418
192
-
 NAD+ pH 9.5, 25°C, recombinant mutant S157E Nostoc sp. PCC 7120 = FACHB-418
300
-
 NAD+ pH 9.5, 25°C, recombinant mutant S157P Nostoc sp. PCC 7120 = FACHB-418