Information on EC 1.2.1.77 - 3,4-dehydroadipyl-CoA semialdehyde dehydrogenase (NADP+)

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The enzyme appears in viruses and cellular organisms

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EC NUMBER
COMMENTARY hide
1.2.1.77
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RECOMMENDED NAME
GeneOntology No.
3,4-dehydroadipyl-CoA semialdehyde dehydrogenase (NADP+)
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3,4-didehydroadipyl-CoA semialdehyde + NADP+ + H2O = 3,4-didehydroadipyl-CoA + NADPH + H+
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
benzoyl-CoA degradation I (aerobic)
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Benzoate degradation
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SYSTEMATIC NAME
IUBMB Comments
3,4-dehydroadipyl-CoA semialdehyde:NADP+ oxidoreductase
This enzyme catalyses a step in the aerobic benzoyl-coenzyme A catabolic pathway in Azoarcus evansii and Burkholderia xenovorans.
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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enzyme catalyzes an essential step in the metabolism of benzoate in Burkholderia xenovorans LB400
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3,4-dehydroadipyl-CoA semialdehyde + NADP+ + H2O
3,4-dehydroadipyl-CoA + NADPH + H+
show the reaction diagram
benzaldehyde + NADP+ + H2O
benzoate + NADPH + H+
show the reaction diagram
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?
cis-3,4-dehydroadipyl-CoA semialdehyde + NADP+ + H2O
cis-3,4-dehydroadipyl-CoA + NADPH + H+
show the reaction diagram
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catalytic mechanism proposed in detail
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?
formaldehyde + NADP+ + H2O
formate + NADPH + H+
show the reaction diagram
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?
heptaldehyde + NADP+ + H2O
heptanoate + NADPH + H+
show the reaction diagram
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?
isovaleraldehyde + NADP+ + H2O
isovalerate + NADPH + H+
show the reaction diagram
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?
propionaldehyde + NADP+ + H2O
propionate + NADPH + H+
show the reaction diagram
valeraldehyde + NADP+ + H2O
valerate + NADPH + H+
show the reaction diagram
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?
additional information
?
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the native substrate (3,4-dehydroadipyl-CoA semialdehyde) is not tested as it is commercially unavailable. The enzyme is preferentially active towards linear medium-chain to long-chain aldehydes as compared to branched-chain, short-chain or aromatic aldehydes
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
3,4-dehydroadipyl-CoA semialdehyde + NADP+ + H2O
3,4-dehydroadipyl-CoA + NADPH + H+
show the reaction diagram
cis-3,4-dehydroadipyl-CoA semialdehyde + NADP+ + H2O
cis-3,4-dehydroadipyl-CoA + NADPH + H+
show the reaction diagram
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catalytic mechanism proposed in detail
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?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
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the enzyme is more active in the presence of NADP+ relative to NAD+. Crystallographic data show that cofactor selectivity is governed by a complex network of hydrogen bonds between the oxygen atoms of the 2'-phosphoryl moiety of NADP+ and a threonine/lysine pair on the enzyme
NADP+
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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salt concentrations up to 500 mM KCl have no effect on enzyme activity
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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salt concentrations up to 500 mM KCl and EDTA concentrations up to 50 mM have no effect on enzyme activity
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.025
3,4-dehydroadipyl-CoA semialdehyde
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pH 7.2
4.15
benzaldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
1.9
formaldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
0.042
heptaldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
1.66
Isovaleraldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
0.501
NAD+
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in 50 mM Tris-HCl (pH 7.5), at 25°C
0.016 - 0.04
NADP+
1.21
propionaldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
0.3
Valeraldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
45
3,4-dehydroadipyl-CoA semialdehyde
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pH 7.2
0.019
benzaldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
0.046
formaldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
29.9
heptaldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
0.074
Isovaleraldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
0.501
NAD+
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in 50 mM Tris-HCl (pH 7.5), at 25°C, cosubstrate: propionaldehyde
0.04 - 45
NADP+
2.32 - 162
propionaldehyde
5.16
Valeraldehyde
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in 50 mM Tris-HCl (pH 7.5), at 25°C
additional information
additional information
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no activities detectable for C296A and E257Q mutant enzymes
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.17
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extracts of cells grown aerobically on benzoate
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2 - 8.8
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50% of maximal activity at pH 6.2 and pH 8.8
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
54000
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2 * 54000
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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forms a dimer in solution with a molecular mass of 110000, SDS-PAGE
dimer
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2 * 54000
homodimer
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
E257Q-NADP+ mutant at 1.4 A resolution, C296A-NADP+ at 1.4 A resolution, E167A mutant at 1.6 A resolution, E496A mutant at 1.65 A resolution
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sitting drop vapour diffusion method, at 18°C in 29% PEG 3350K and 100 mM bis-Tris (pH 6.0), 1.6 A crystal structure of ALDHC in complex with NADPH bound in the cofactor-binding pocket and an ordered fragment of a polyethylene glycol molecule bound in the substrate tunnel
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, enzyme could be stored without appreciable loss of activity for months at -20°C in the presence of 10% (v/v) glycerol
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by nickel affinity column and gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21 Star (DE3) cells
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expression in Escherichia coli as a protein tagged at its N terminus with maltose-binding protein
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mutations confirmed by sequence analysis
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C296A
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same purification behavior as the native enzyme indicating that the mutation does not alter basic enzyme structure, but no catalytic activity detected consistent with a critical nucleophilic role for C296
E167A
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significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 77%, likely candidate to present the catalytic water to the general base E257 that leads to deacylation and product release
E257Q
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tolerated with respect to overall protein stability, but dramatic reduction in kcat value and no activity detectable, E257 serves probably as the primary general base to deprotonate the nucleophilic C296
E400A
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91% reduction in kcat value, E400 plays an essential role in facilitating the acquisition of the hydride conformation
E496A
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significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 90%
H485A
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dramatic reduction in enzyme solubility
H485Q
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completely recalcitrant to purification, H485 may be important in stabilizing the nicotinamide amide moiety of NADP+
K168A
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significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 92.5%, K168 likely serves to stabilize charges and tune pKa of the active site glutamate side-chains
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LINKS TO OTHER DATABASES (specific for EC-Number 1.2.1.77)
ExplorEnz
ExPASy
KEGG
MetaCyc
SABIO-RK
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
InterPro (database of protein families, domains and functional sites)