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Literature summary for 1.2.1.87 extracted from

  • Baker, P.; Pan, D.; Carere, J.; Rossi, A.; Wang, W.; Seah, S.Y.K.
    Characterization of an aldolase-dehydrogenase complex that exhibits substrate channeling in the polychlorinated biphenyls degradation pathway (2009), Biochemistry, 48, 6551-6558.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
coexpression of bphI and bphJ in Escherichia coli using two compatible plasmids (pBTL4 and pET28a) yield soluble proteins Paraburkholderia xenovorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0348
-
NAD+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
0.561
-
NADP+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
7.7
-
Isobutyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
23.1
-
propionaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
23.6
-
acetaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
31.7
-
Butyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
SDS-PAGE, in agreement with the predicted molecular mass calculated from amino acid sequence Paraburkholderia xenovorans
37000
-
BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI, estimated by gel filtration Paraburkholderia xenovorans
140000
-
native molecular mass of the purified BphI-BphJ-complex, estimated by gel filtration Paraburkholderia xenovorans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
propanal + CoA + NAD+ Paraburkholderia xenovorans
-
propanoyl-CoA + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Paraburkholderia xenovorans Q79AF6
-
-

Purification (Commentary)

Purification (Comment) Organism
BphI and BphJ form a stable complex as they bind and coelute from Ni2+-NTA column, although only BphJ has the histidine tag. After purification, the N-terminal histidine tag of BphJ is proteolytically cleaved by thrombin digestion Paraburkholderia xenovorans

Storage Stability

Storage Stability Organism
purified enzyme can be stored at -80°C, without loss of activity for at least 12 months Paraburkholderia xenovorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetaldehyde + CoA + NAD+
-
Paraburkholderia xenovorans acetyl-CoA + NADH + H+
-
?
acetaldehyde + CoA + NADP+ BphJ is able to utilize NAD+ and NADP+ with comparable kcat values, but the apparent Km-value for NAD+ is 16fold lower than for NADP+ Paraburkholderia xenovorans acetyl-CoA + NADPH + H+
-
?
butyraldehyde + CoA + NAD+
-
Paraburkholderia xenovorans butyryl-CoA + NADH + H+
-
?
isobutyraldehyde + CoA + NAD+
-
Paraburkholderia xenovorans isobutyryl-CoA + NADH + H+
-
?
additional information no activity with succinic semialdehyde and picolinaldehyde Paraburkholderia xenovorans ?
-
?
pentaldehyde + CoA + NAD+
-
Paraburkholderia xenovorans pentanoyl-CoA + NADH + H+
-
?
propanal + CoA + NAD+
-
Paraburkholderia xenovorans propanoyl-CoA + NADH + H+
-
?
propionaldehyde + CoA + NAD+
-
Paraburkholderia xenovorans propanoyl-CoA + NADH + H+
-
?

Subunits

Subunits Comment Organism
heterotetramer BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI, estimated by gel filtration Paraburkholderia xenovorans

Synonyms

Synonyms Comment Organism
BphJ
-
Paraburkholderia xenovorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
9.5
-
Butyraldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
11.2
-
Isobutyraldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
12.9
-
NADP+ with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
14.1
-
NAD+ with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
16.3
-
propionaldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
17.2
-
acetaldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans

Cofactor

Cofactor Comment Organism Structure
NAD+ BphJ is able to utilize NAD+ and NADP+ with comparable kcat values, but the apparent Km-value for NAD+ is 16fold lower than for NADP+ Paraburkholderia xenovorans
NADP+ BphJ is able to utilize NAD+ and NADP+ with comparable kcat values, but the apparent Km-value for NAD+ is 16fold lower than for NADP+ Paraburkholderia xenovorans

General Information

General Information Comment Organism
metabolism enzyme forms a bifunctional complex with EC 4.1.3.43 (BphI), 4-hydroxy-2-oxohexanoate aldolase and is part of the polychlorinated biphenyl degradation pathway Paraburkholderia xenovorans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.076
-
pentaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
0.3
-
Butyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
0.7
-
propionaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
0.73
-
acetaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.4
-
Isobutyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
22.99
-
NADP+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
405.2
-
NAD+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans