A5JTM5 · CBADH_PSEUC
- Protein4-chlorobenzoyl coenzyme A dehalogenase
- StatusUniProtKB reviewed (Swiss-Prot)
- Organism
- Amino acids269 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
Dehalogenates 4-chlorobenzoyl-CoA, 4-iodobenzoyl-CoA and 4-bromobenzoyl-CoA, but not 4-fluorobenzoyl-CoA. Inactive towards crotonyl-CoA, alpha-methylcrotonyl-CoA and beta-methylcrotonyl-CoA.
Catalytic activity
- 4-chlorobenzoyl-CoA + H2O = 4-hydroxybenzoyl-CoA + chloride + H+
Activity regulation
Inactivated by 1 mM Ag+ and by 5 mM Cu2+. Partially inhibited by 5 mM Zn2+, Mn2+, Co2+, Fe2+ and Ni2+. Unaffected by 10 mM Na+, K+ and Li+ and by 0.5 mM Mg2+, Mn2+, Fe2+, Ca2+, Co2+ and Zn2+. Inhibited by the sulfhydryl blocking agent 5,5'-dithio-bis-(2-nitrobenzoate), SDS and N-bromosuccinimide. Unaffected by sodium azide and EDTA. Inactivated following treatment with diethyl pyrocarbonate; this inactivation is reversible by treatment with hydroxylamine.
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
3.7 μM | 4-chlorobenzoyl-CoA | |||||
4.2 μM | 4-bromobenzoyl-CoA | |||||
6.5 μM | 4-iodobenzoyl-CoA | |||||
10.4 μM | 2,4-dichlorobenzoyl-CoA | |||||
42 μM | 3,4-dichlorobenzoyl-CoA | |||||
30 μM | 4-chloro-2-nitrobenzoyl-CoA | |||||
5.5 μM | 4-chloro-3-nitrobenzoyl-CoA |
Vmax | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|
2.2 μmol/min/mg | with 4-chlorobenzoyl-CoA as substrate |
pH Dependence
Optimum pH is 10.0. Half maximum activity remains at pH 9.0 and 11.5. Stable from pH 6.5 to 11.0, activity is lost following 10 minutes incubation at pH 4.5 or 12.0.
Temperature Dependence
Optimum temperature is 60 degrees Celsius. Retains 60% of maximum activity at 30 degrees Celsius and 65 degrees Celsius. After 15 minutes preincubation at 60 degrees Celsius 70% of the initial activity remains, 15 minutes preincubation at 65 degrees Celsius results in a total loss of activity.
Pathway
Xenobiotic degradation; 4-chlorobenzoate degradation; 4-hydroxybenzoate from 4-chlorobenzoate: step 2/3.
Features
Showing features for binding site, active site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Binding site | 24 | substrate; ligand shared between two oligomeric partners; in other chain | ||||
Sequence: R | ||||||
Binding site | 62-67 | substrate; ligand shared between two oligomeric partners; in other chain | ||||
Sequence: AGFYLR | ||||||
Active site | 90 | Proton acceptor | ||||
Sequence: H | ||||||
Binding site | 114 | substrate; ligand shared between two oligomeric partners; in other chain | ||||
Sequence: G | ||||||
Active site | 145 | Nucleophile | ||||
Sequence: D | ||||||
Binding site | 257 | substrate; ligand shared between two oligomeric partners | ||||
Sequence: R |
GO annotations
Aspect | Term | |
---|---|---|
Molecular Function | 4-chlorobenzoyl-CoA dehalogenase activity | |
Biological Process | coenzyme A metabolic process |
Keywords
- Molecular function
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended name4-chlorobenzoyl coenzyme A dehalogenase
- EC number
- Short names4-CBA-CoA dehalogenase ; 4-CBCoA dehalogenase ; 4-chlorobenzoyl-CoA dehalogenase
Organism names
- Organism
- Strain
- Taxonomic lineageBacteria > Pseudomonadota
Accessions
- Primary accessionA5JTM5
Phenotypes & Variants
Features
Showing features for mutagenesis.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Mutagenesis | 24 | Does not strongly affect catalytic activity, but reduces substrate CoA binding. | ||||
Sequence: R → K or L | ||||||
Mutagenesis | 34 | Forms inclusion bodies. | ||||
Sequence: E → T | ||||||
Mutagenesis | 43 | No effect on catalytic activity. | ||||
Sequence: E → A | ||||||
Mutagenesis | 45 | No effect on catalytic activity. | ||||
Sequence: D → A | ||||||
Mutagenesis | 46 | No effect on catalytic activity. | ||||
Sequence: D → A | ||||||
Mutagenesis | 63 | Yields insoluble protein. | ||||
Sequence: G → A, I, or P | ||||||
Mutagenesis | 64 | 30-fold reduction in catalytic activity, substrate benzoyl group binding is unaffected. | ||||
Sequence: F → A | ||||||
Mutagenesis | 64 | Retains catalytic activity, but substrate benzoyl group binding is decreased. | ||||
Sequence: F → L | ||||||
Mutagenesis | 64 | Severely reduces catalytic activity. Arylated intermediate does not accumulate. | ||||
Sequence: F → P | ||||||
Mutagenesis | 65 | Catalytic activity is almost as efficient as wild type. | ||||
Sequence: Y → D | ||||||
Mutagenesis | 67 | Reduces substrate CoA binding. | ||||
Sequence: R → K | ||||||
Mutagenesis | 67 | Forms inclusion bodies. | ||||
Sequence: R → L | ||||||
Mutagenesis | 68 | No effect on catalytic activity. | ||||
Sequence: E → T | ||||||
Mutagenesis | 81 | Loss of catalytic activity, substrate benzoyl group binding is not affected. | ||||
Sequence: H → Q | ||||||
Mutagenesis | 82 | Retains catalytic activity, but substrate benzoyl group binding is decreased. | ||||
Sequence: F → L | ||||||
Mutagenesis | 89 | Retains catalytic activity, but substrate benzoyl group binding is decreased. | ||||
Sequence: W → F | ||||||
Mutagenesis | 89 | Reduced activity and substrate benzoyl group binding. | ||||
Sequence: W → Y | ||||||
Mutagenesis | 90 | Complete loss of catalytic activity (PubMed:8718880, PubMed:9063883). Significantly reduced activity (PubMed:11695894). Substrate binding is not significantly affected. Reduced arylated intermediate formation. | ||||
Sequence: H → Q | ||||||
Mutagenesis | 94 | No effect on catalytic activity. | ||||
Sequence: H → Q | ||||||
Mutagenesis | 112 | Yields insoluble protein. | ||||
Sequence: A → G | ||||||
Mutagenesis | 112 | Protein precipitates upon purification. | ||||
Sequence: A → S | ||||||
Mutagenesis | 112 | Catalytic activity is almost as efficient as wild type. | ||||
Sequence: A → V | ||||||
Mutagenesis | 113 | Strongly reduced catalytic activity and substrate benzoyl group binding. Arylated intermediate does not accumulate. | ||||
Sequence: G → A | ||||||
Mutagenesis | 113 | Strongly reduced catalytic activity. Arylated intermediate does not accumulate. | ||||
Sequence: G → N or S | ||||||
Mutagenesis | 113 | Protein precipitates upon purification. | ||||
Sequence: G → V | ||||||
Mutagenesis | 114 | Strongly reduced catalytic activity and substrate benzoyl group binding. | ||||
Sequence: G → A | ||||||
Mutagenesis | 114 | Unstable. | ||||
Sequence: G → P | ||||||
Mutagenesis | 115 | Yields insoluble protein. | ||||
Sequence: G → L, N, S, or V | ||||||
Mutagenesis | 123 | No effect on catalytic activity. | ||||
Sequence: D → T | ||||||
Mutagenesis | 129 | No effect on catalytic activity. | ||||
Sequence: D → T | ||||||
Mutagenesis | 137 | Low catalytic activity, but KM unaffected (PubMed:8718880). Retains catalytic activity, but substrate benzoyl group binding is decreased (PubMed:9063883). | ||||
Sequence: W → F | ||||||
Mutagenesis | 145 | Complete loss of catalytic activity, but not substrate binding. | ||||
Sequence: D → A | ||||||
Mutagenesis | 163 | No effect on catalytic activity. | ||||
Sequence: E → T | ||||||
Mutagenesis | 175 | No effect on catalytic activity. | ||||
Sequence: E → D | ||||||
Mutagenesis | 179 | No effect on catalytic activity. | ||||
Sequence: W → F | ||||||
Mutagenesis | 208 | No effect on catalytic activity. | ||||
Sequence: H → Q | ||||||
Mutagenesis | 216 | Yields insoluble protein. | ||||
Sequence: R → E, K, or L | ||||||
Mutagenesis | 232 | Yields insoluble protein. | ||||
Sequence: E → A, N, Q, or R | ||||||
Mutagenesis | 232 | Reduced catalytic activity, increased substrate binding. | ||||
Sequence: E → D | ||||||
Mutagenesis | 257 | Retains catalytic activity and substrate CoA binding. | ||||
Sequence: R → K | ||||||
Mutagenesis | 257 | Significantly reduces catalytic activity and substrate CoA binding. | ||||
Sequence: R → L |
PTM/Processing
Features
Showing features for chain.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Chain | PRO_0000401147 | 1-269 | 4-chlorobenzoyl coenzyme A dehalogenase | |||
Sequence: MYEAIGHRVEDGVAEITIKLPRHRNALSVKAMQEVTDALNRAEEDDSVGAVMITGAEDAFCAGFYLREIPLDKGVAGVRDHFRIGALWWHQMIHKIIRVKRPVLAAINGVAAGGGLGISLASDMAICADSAKFVCAWHTIGIGNDTATSYSLARIVGMRRAMELMLTNRTLYPEEAKDWGLVSRVYPKDDFREVAWKVARELAAAPTHLQVMAKERFHAGWMQPVEECTEFEIQNVIASVTHPHFMPCLTEFLDGHRADRPQVELPAGV |
Expression
Induction
By 4-chlorobenzoyl-CoA, 4-iodobenzoyl-CoA or 4-bromobenzoyl-CoA.
Structure
Sequence
- Sequence statusComplete
- Length269
- Mass (Da)29,802
- Last updated2007-06-26 v1
- ChecksumEC165E9063C70B07
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
EF569604 EMBL· GenBank· DDBJ | ABQ44578.1 EMBL· GenBank· DDBJ | Genomic DNA |