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Literature summary 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

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

Cloned(Commentary)

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

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.87 0.0348
-
 NAD+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
1.2.1.87 0.561
-
 NADP+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
1.2.1.87 7.7
-
 Isobutyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 23.1
-
 propionaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 23.6
-
 acetaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 31.7
-
 Butyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
4.1.3.43 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
4.1.3.43 0.18
-
 4-hydroxy-2-oxohexanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.22
-
 4-hydroxy-2-oxopentanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.35
-
 4-hydroxy-2-oxoheptanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 7.5
-
 oxaloacetate pH 8.0, 25°C Paraburkholderia xenovorans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.2.1.87 32000
-
 SDS-PAGE, in agreement with the predicted molecular mass calculated from amino acid sequence Paraburkholderia xenovorans
1.2.1.87 37000
-
 BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI, estimated by gel filtration Paraburkholderia xenovorans
1.2.1.87 140000
-
 native molecular mass of the purified BphI-BphJ-complex, estimated by gel filtration Paraburkholderia xenovorans
4.1.3.43 32000
-
 BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI Paraburkholderia xenovorans
4.1.3.43 37000
-
 SDS-PAGE, in agreement with the predicted molecular mass calculated from amino acid sequence Paraburkholderia xenovorans
4.1.3.43 140000
-
 native molecular mass of the purified BphI-BphJ-complex, estimated by gel filtration Paraburkholderia xenovorans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.1.87 propanal + CoA + NAD+ Paraburkholderia xenovorans
-
 propanoyl-CoA + NADH + H+
-
 ?
4.1.3.43 (S)-4-hydroxy-2-oxohexanoate Paraburkholderia xenovorans
-
 propanal + pyruvate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.87 Paraburkholderia xenovorans Q79AF6
-
-
4.1.3.43 Paraburkholderia xenovorans P51015
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.87 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
4.1.3.43 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

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.87 acetaldehyde + CoA + NAD+
-
 Paraburkholderia xenovorans acetyl-CoA + NADH + H+
-
 ?
1.2.1.87 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+
-
 ?
1.2.1.87 butyraldehyde + CoA + NAD+
-
 Paraburkholderia xenovorans butyryl-CoA + NADH + H+
-
 ?
1.2.1.87 isobutyraldehyde + CoA + NAD+
-
 Paraburkholderia xenovorans isobutyryl-CoA + NADH + H+
-
 ?
1.2.1.87 additional information no activity with succinic semialdehyde and picolinaldehyde Paraburkholderia xenovorans ?
-
 ?
1.2.1.87 pentaldehyde + CoA + NAD+
-
 Paraburkholderia xenovorans pentanoyl-CoA + NADH + H+
-
 ?
1.2.1.87 propanal + CoA + NAD+
-
 Paraburkholderia xenovorans propanoyl-CoA + NADH + H+
-
 ?
1.2.1.87 propionaldehyde + CoA + NAD+
-
 Paraburkholderia xenovorans propanoyl-CoA + NADH + H+
-
 ?
4.1.3.43 (S)-4-hydroxy-2-oxohexanoate
-
 Paraburkholderia xenovorans propanal + pyruvate
-
 ?
4.1.3.43 4-hydroxy-2-oxoheptanoate
-
 Paraburkholderia xenovorans pyruvate + butanal
-
 ?
4.1.3.43 4-hydroxy-2-oxohexanoate
-
 Paraburkholderia xenovorans pyruvate + propionaldehyde
-
 ?
4.1.3.43 4-hydroxy-2-oxopentanoate
-
 Paraburkholderia xenovorans pyruvate + acetaldehyde
-
 ?
4.1.3.43 Oxaloacetate
-
 Paraburkholderia xenovorans Pyruvate + CO2
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
1.2.1.87 BphJ
-
 Paraburkholderia xenovorans
4.1.3.43 BphI
-
 Paraburkholderia xenovorans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.87 9.5
-
 Butyraldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 11.2
-
 Isobutyraldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 12.9
-
 NADP+ with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
1.2.1.87 14.1
-
 NAD+ with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
1.2.1.87 16.3
-
 propionaldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 17.2
-
 acetaldehyde pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
4.1.3.43 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
4.1.3.43 0.63
-
 4-hydroxy-2-oxoheptanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 3.8
-
 4-hydroxy-2-oxohexanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 4.1
-
 4-hydroxy-2-oxopentanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 16.5
-
 oxaloacetate pH 8.0, 25°C Paraburkholderia xenovorans

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.87 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
1.2.1.87 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

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.1.3.43 0.00093
-
 oxalate pH 8.0, 25°C Paraburkholderia xenovorans

General Information

EC Number General Information Comment Organism
1.2.1.87 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
4.1.3.43 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.2.1.87 0.076
-
 pentaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 0.3
-
 Butyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 0.7
-
 propionaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 0.73
-
 acetaldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 1.4
-
 Isobutyraldehyde app. Km-value, pH 8.0 and 25°C, NAD+ Paraburkholderia xenovorans
1.2.1.87 22.99
-
 NADP+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
1.2.1.87 405.2
-
 NAD+ app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C Paraburkholderia xenovorans
4.1.3.43 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
4.1.3.43 1.8
-
 4-hydroxy-2-oxoheptanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 2.2
-
 oxaloacetate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 18.6
-
 4-hydroxy-2-oxopentanoate pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 21.1
-
 4-hydroxy-2-oxohexanoate pH 8.0, 25°C Paraburkholderia xenovorans