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

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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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.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
enzyme catalyzes an essential step in the metabolism of benzoate in Burkholderia xenovorans LB400
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
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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-
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?
isovaleraldehyde + NADP+ + H2O
isovalerate + NADPH + H+
show the reaction diagram
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-
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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
Q13WK4
catalytic mechanism proposed in detail
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?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
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+
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
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
in 50 mM Tris-HCl (pH 7.5), at 25C
1.9
formaldehyde
in 50 mM Tris-HCl (pH 7.5), at 25C
0.042
heptaldehyde
in 50 mM Tris-HCl (pH 7.5), at 25C
1.66
Isovaleraldehyde
in 50 mM Tris-HCl (pH 7.5), at 25C
0.501
NAD+
in 50 mM Tris-HCl (pH 7.5), at 25C
0.016 - 0.04
NADP+
1.21
propionaldehyde
in 50 mM Tris-HCl (pH 7.5), at 25C
0.3
Valeraldehyde
in 50 mM Tris-HCl (pH 7.5), at 25C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
45
3,4-dehydroadipyl-CoA semialdehyde
Azoarcus evansii
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pH 7.2
0.019
benzaldehyde
Paraburkholderia xenovorans
Q13WK4
in 50 mM Tris-HCl (pH 7.5), at 25C
0.046
formaldehyde
Paraburkholderia xenovorans
Q13WK4
in 50 mM Tris-HCl (pH 7.5), at 25C
29.9
heptaldehyde
Paraburkholderia xenovorans
Q13WK4
in 50 mM Tris-HCl (pH 7.5), at 25C
0.074
Isovaleraldehyde
Paraburkholderia xenovorans
Q13WK4
in 50 mM Tris-HCl (pH 7.5), at 25C
0.501
NAD+
Paraburkholderia xenovorans
Q13WK4
in 50 mM Tris-HCl (pH 7.5), at 25C, cosubstrate: propionaldehyde
0.04 - 45
NADP+
2.32 - 162
propionaldehyde
5.16
Valeraldehyde
Paraburkholderia xenovorans
Q13WK4
in 50 mM Tris-HCl (pH 7.5), at 25C
additional information
additional information
Paraburkholderia xenovorans
Q13WK4
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
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
forms a dimer in solution with a molecular mass of 110000, SDS-PAGE
dimer
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2 * 54000
homodimer
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
sitting drop vapour diffusion method, at 18C 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
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, enzyme could be stored without appreciable loss of activity for months at -20C in the presence of 10% (v/v) glycerol
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by nickel affinity column and gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21 Star (DE3) cells
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
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C296A
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
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
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
91% reduction in kcat value, E400 plays an essential role in facilitating the acquisition of the hydride conformation
E496A
significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 90%
H485A
dramatic reduction in enzyme solubility
H485Q
completely recalcitrant to purification, H485 may be important in stabilizing the nicotinamide amide moiety of NADP+
K168A
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
Show AA Sequence (154 entries)
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