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Literature summary extracted from

  • Carere, J.; Baker, P.; Seah, S.Y.K.
    Investigating the molecular determinants for substrate channeling in BphI-BphJ, an aldolase-dehydrogenase complex from the polychlorinated biphenyls degradation pathway (2011), Biochemistry, 50, 8407-8416.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.87 bphI and bphJ cloned into the plasmids pBTL4-T7 and pET28a Paraburkholderia xenovorans
4.1.3.43 bphI and bphJ cloned into the plasmids pBTL4-T7 and pET28a Paraburkholderia xenovorans

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.87 I195F created by site-specific mutagenesis, no significant reduction of channeling efficiency of the enzyme complex toward acetaldehyde or propionaldehyde Paraburkholderia xenovorans
1.2.1.87 I195L created by site-specific mutagenesis Paraburkholderia xenovorans
4.1.3.43 G322A channels acetaldehyde with similar efficiency to wild-type, 30% lowered efficiency of isobutyraldehyde channeling Paraburkholderia xenovorans
4.1.3.43 G322F unable to channel either acetaldehyde or propionaldehyde Paraburkholderia xenovorans
4.1.3.43 G322L unable to channel either acetaldehyde or propionaldehyde Paraburkholderia xenovorans
4.1.3.43 G323A channels acetaldehyde with similar efficiency to wild-type, unable to channel isobutyraldehyde Paraburkholderia xenovorans
4.1.3.43 G323F unable to channel either acetaldehyde or propionaldehyde Paraburkholderia xenovorans
4.1.3.43 G323L 63% channeling efficiency for acetaldehyde, unable to channel propionaldehyde Paraburkholderia xenovorans
4.1.3.43 H20A reduction of acetaldehyde and propionaldehyde channeling by more than 70% Paraburkholderia xenovorans
4.1.3.43 L89A reduction in channeling efficiency of 30% for all aldehydes tested Paraburkholderia xenovorans
4.1.3.43 additional information all mutations created by site specific mutagenesis Paraburkholderia xenovorans
4.1.3.43 Y290F reduction in channeling efficiency of 30% for all aldehydes tested Paraburkholderia xenovorans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.87 2 3 propionaldehyde wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 7.4
-
 acetaldehyde I195L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 7.7
-
 propionaldehyde I195L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 24
-
 acetaldehyde wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 35
-
 acetaldehyde I195F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 48
-
 propionaldehyde I195F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.053
-
 4-hydroxy-2-oxopentanoate G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.06
-
 4-hydroxy-2-oxopentanoate G323A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.081
-
 4-hydroxy-2-oxopentanoate G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.089
-
 4-hydroxy-2-oxopentanoate wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.09
-
 4-hydroxy-2-oxopentanoate G322F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.117
-
 4-hydroxy-2-oxohexanoate wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.13
-
 4-hydroxy-2-oxopentanoate G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.14
-
 4-hydroxy-2-oxopentanoate G322L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.158
-
 4-hydroxy-2-oxopentanoate wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.16
-
 4-hydroxy-2-oxopentanoate G323L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.18
-
 4-hydroxy-2-oxopentanoate G322A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.216
-
 4-hydroxy-2-oxopentanoate G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.3
-
 4-hydroxy-2-oxohexanoate G322F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.35
-
 4-hydroxy-2-oxohexanoate G322L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.36
-
 4-hydroxy-2-oxohexanoate G323A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.37
-
 4-hydroxy-2-oxohexanoate G323L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.52
-
 4-hydroxy-2-oxohexanoate G322A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.94
-
 4-hydroxy-2-oxopentanoate G323F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 1.1
-
 4-hydroxy-2-oxohexanoate G323F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans

Organism

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

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, after purification 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, after purification N-terminal histidine tag of BphJ is proteolytically cleaved by thrombin digestion 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 propionaldehyde + CoA + NAD+
-
 Paraburkholderia xenovorans propanoyl-CoA + NADH + H+
-
 ?
4.1.3.43 4-hydroxy-2-oxohexanoate 4-hydroxy-2-oxohexanoate i.e. HOHA Paraburkholderia xenovorans propionaldehyde + pyruvate
-
 ?
4.1.3.43 4-hydroxy-2-oxopentanoate 4-hydroxy-2-oxopentanoate i.e. HOPA Paraburkholderia xenovorans acetaldehyde + pyruvate
-
 ?

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 8.9
-
 propionaldehyde I195F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 16
-
 acetaldehyde I195F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 16.3
-
 propionaldehyde wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 17.2
-
 acetaldehyde wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 42
-
 acetaldehyde I195L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
1.2.1.87 46
-
 propionaldehyde I195L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.1
-
 4-hydroxy-2-oxopentanoate G323L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.27
-
 4-hydroxy-2-oxohexanoate G323L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.286
-
 4-hydroxy-2-oxopentanoate G322L mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.34
-
 4-hydroxy-2-oxopentanoate G323A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.45
-
 4-hydroxy-2-oxopentanoate G322F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.46
-
 4-hydroxy-2-oxohexanoate G323A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.64
-
 4-hydroxy-2-oxopentanoate G323L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.72
-
 4-hydroxy-2-oxopentanoate G323F mutant, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.79
-
 4-hydroxy-2-oxopentanoate wild-type protein, steady state, absence of BphJ cofactors, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 0.94
-
 4-hydroxy-2-oxohexanoate G322L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 1.05
-
 4-hydroxy-2-oxohexanoate G322F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 1.06
-
 4-hydroxy-2-oxopentanoate G322F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 1.09
-
 4-hydroxy-2-oxopentanoate G322L mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 1.3
-
 4-hydroxy-2-oxohexanoate G323F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 1.9
-
 4-hydroxy-2-oxohexanoate G322A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 2.4
-
 4-hydroxy-2-oxopentanoate G322A mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 3.9
-
 4-hydroxy-2-oxohexanoate wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 4.07
-
 4-hydroxy-2-oxopentanoate wild-type protein, steady state, pH 8.0, 25°C Paraburkholderia xenovorans
4.1.3.43 4.3
-
 4-hydroxy-2-oxopentanoate G323F mutant, steady state, pH 8.0, 25°C Paraburkholderia xenovorans

General Information

EC Number General Information Comment Organism
1.2.1.87 metabolism enzyme is part of the BphI-BphJ-complex, an aldolase-dehydrogenase-complex from the polychlorinated biphenyl degradation pathway Paraburkholderia xenovorans
4.1.3.43 metabolism enzyme is part of the BphI-BphJ-complex, an aldolase-dehydrogenase complex from the polychlorinated biphenyl degradation pathway Paraburkholderia xenovorans