Crystallization (Comment) | Organism |
---|---|
OCH mutant H122A in complex with the minor diastereoisomer of (2S,4S)-alpha-campholinic acid, vapor diffusion hanging drop technique, mxing of protein solution containing 10 mg/ml protein in 50 mM Tris-HCl, pH 7.1, 1 mM dithiothreitol, and 0.02 mM phenylmethylsulfonyl fluoride with reservoir solution containing 0.1 M 2-(N-morpholino)ethanesulfonic acid, pH 5.6, 0.2 M calcium acetate, and 26% v/v PEG monomethyl ether 2000, in a 1:1 ratio, soakong of crystals in reservoir solution containing 6-oxocamphor for 30 min, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement | Rhodococcus sp. |
Protein Variants | Comment | Organism |
---|---|---|
D154N | site-directed mutagenesis, the OCH mutant shows a greatly reduced value of kcat/Km derived from a very large increase in Km for the native substrate 6-oxo camphor compared to the wild-type enzyme | Rhodococcus sp. |
E244Q | site-directed mutagenesis, the mutant shows reduced value of kcat/Km compared to the wild-type enzyme | Rhodococcus sp. |
H122A | site-directed mutagenesis, structure analysis and comparison to the wild-type enzyme, the mutant shows a greatly reduced value of kcat, and its Km is five times that of the wild-type enzyme. The H122A mutant forms a hexamer, a dimer of trimers, identical to that of the wild-type enzyme | Rhodococcus sp. |
H145A | site-directed mutagenesis, the OCH mutant shows a greatly reduced value of kcat/Km derived from a very large increase in Km for the native substrate 6-oxo camphor compared to the wild-type enzyme | Rhodococcus sp. |
H45A | site-directed mutagenesis, the mutant shows reduced value of kcat/Km compared to the wild-type enzyme | Rhodococcus sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analysis of OCH mutants, overview | Rhodococcus sp. | |
0.04 | - |
6-oxocamphor | wild-type enzyme, pH and temperature not specified in the publication | Rhodococcus sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
6-oxocamphor + H2O | Rhodococcus sp. | - |
(2S,4S)-alpha-campholinic acid | - |
? | |
6-oxocamphor + H2O | Rhodococcus sp. NCIMB 9784 | - |
(2S,4S)-alpha-campholinic acid | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | Q93TU6 | - |
- |
Rhodococcus sp. NCIMB 9784 | Q93TU6 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
bornane-2,6-dione + H2O = [(1S)-4-hydroxy-2,2,3-trimethylcyclopent-3-enyl]acetate | involvement of five residues, His45, His122, His145, Asp154, and Glu244, in catalysis, catalytic His145/Asp154 dyad. The the pendant acetate of the product (2S,4S)-alpha-campholinic acid hydrogen bonded to a His145/Asp154 dyad and the endocyclic carbonyl of the cyclopentane ring hydrogen bonds to Trp40, prochiral selectivity, base-catalyzed mechanism of C-C bond cleavage, overview | Rhodococcus sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
6-oxocamphor + H2O | - |
Rhodococcus sp. | (2S,4S)-alpha-campholinic acid | - |
? | |
6-oxocamphor + H2O | - |
Rhodococcus sp. NCIMB 9784 | (2S,4S)-alpha-campholinic acid | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | dimer of trimers, crystal structure, overview | Rhodococcus sp. |
Synonyms | Comment | Organism |
---|---|---|
6-oxo camphor hydrolase | - |
Rhodococcus sp. |
OCH | - |
Rhodococcus sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
642.9 | - |
6-oxocamphor | wild-type enzyme, pH and temperature not specified in the publication | Rhodococcus sp. |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the crotonase superfamily | Rhodococcus sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
16100 | - |
6-oxocamphor | wild-type enzyme, pH and temperature not specified in the publication | Rhodococcus sp. |