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Literature summary for 4.1.3.43 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

Activating Compound

Activating Compound Comment Organism Structure
BphJ 15fold increased activity Paraburkholderia xenovorans

Cloned(Commentary)

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

Inhibitors

Inhibitors Comment Organism Structure
Cd2+ 56.0% activity at 0.1 mM and 5.4% activity at 1 mM chloride salt Paraburkholderia xenovorans
Co2+ 19.3% activity at 0.1 mM and 18.3% activity at 1 mM chloride salt Paraburkholderia xenovorans
Mg2+ 3.4% activity at 0.1 mM and 14.8% activity at 1 mM chloride salt Paraburkholderia xenovorans
Mn2+ 100% activity at 0.1 mM and 1 mM chloride salt Paraburkholderia xenovorans
Ni2+ 2.6% activity at 0.1 mM and 17.5% activity at 1 mM chloride salt Paraburkholderia xenovorans
oxalate
-
Paraburkholderia xenovorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information presence of NAD+, NADP+, NADH or CoA leads to an at least 5fold reduction in the Km for 4-hydroxy-2-oxopentanoate Paraburkholderia xenovorans
0.18
-
4-hydroxy-2-oxohexanoate pH 8.0, 25°C Paraburkholderia xenovorans
0.22
-
4-hydroxy-2-oxopentanoate pH 8.0, 25°C Paraburkholderia xenovorans
0.35
-
4-hydroxy-2-oxoheptanoate pH 8.0, 25°C Paraburkholderia xenovorans
7.5
-
oxaloacetate pH 8.0, 25°C Paraburkholderia xenovorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI Paraburkholderia xenovorans
37000
-
SDS-PAGE, in agreement with the predicted molecular mass calculated from amino acid sequence 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.
(S)-4-hydroxy-2-oxohexanoate Paraburkholderia xenovorans
-
propanal + pyruvate
-
?

Organism

Organism UniProt Comment Textmining
Paraburkholderia xenovorans P51015
-
-

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.
(S)-4-hydroxy-2-oxohexanoate
-
Paraburkholderia xenovorans propanal + pyruvate
-
?
4-hydroxy-2-oxoheptanoate
-
Paraburkholderia xenovorans pyruvate + butanal
-
?
4-hydroxy-2-oxohexanoate
-
Paraburkholderia xenovorans pyruvate + propionaldehyde
-
?
4-hydroxy-2-oxopentanoate
-
Paraburkholderia xenovorans pyruvate + acetaldehyde
-
?
Oxaloacetate
-
Paraburkholderia xenovorans Pyruvate + CO2
-
?

Subunits

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

Synonyms

Synonyms Comment Organism
BphI
-
Paraburkholderia xenovorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information 2 and 5fold increases in kcat observed when BphI reaction is conducted in the presence of NAD+ and NADH Paraburkholderia xenovorans
0.63
-
4-hydroxy-2-oxoheptanoate pH 8.0, 25°C Paraburkholderia xenovorans
3.8
-
4-hydroxy-2-oxohexanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1
-
4-hydroxy-2-oxopentanoate pH 8.0, 25°C Paraburkholderia xenovorans
16.5
-
oxaloacetate pH 8.0, 25°C Paraburkholderia xenovorans

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.00093
-
oxalate pH 8.0, 25°C Paraburkholderia xenovorans

General Information

General Information Comment Organism
metabolism enzyme forms a bifunctional complex with EC 1.2.1.87 (BphJ), propanal dehydrogenase (CoA-propanoylating) 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
additional information
-
additional information highest catalytic efficiency of BphI occurs when NADH is present, which is 25fold higher than the value obtained without nucleotides Paraburkholderia xenovorans
1.8
-
4-hydroxy-2-oxoheptanoate pH 8.0, 25°C Paraburkholderia xenovorans
2.2
-
oxaloacetate pH 8.0, 25°C Paraburkholderia xenovorans
18.6
-
4-hydroxy-2-oxopentanoate pH 8.0, 25°C Paraburkholderia xenovorans
21.1
-
4-hydroxy-2-oxohexanoate pH 8.0, 25°C Paraburkholderia xenovorans