Literature summary for 3.7.1.18 extracted from

Leonard, P.; Grogan, G.
Structure of 6-oxo camphor hydrolase H122A mutant bound to its natural product, (2S,4S)-alpha-campholinic acid. Mutant structure suggests an atypical mode of transition state binding for a crotonase homolog (2004), J. Biol. Chem., 279, 31312-31317.

Crystallization (Commentary)

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.

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

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 [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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Rhodococcus sp.	Q93TU6	-	-
Rhodococcus sp. NCIMB 9784	Q93TU6	-	-

Reaction

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.	a

Substrates and Products (Substrate)

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

Subunits	Comment	Organism
hexamer	dimer of trimers, crystal structure, overview	Rhodococcus sp.

Synonyms

Synonyms	Comment	Organism
6-oxo camphor hydrolase	-	Rhodococcus sp.
OCH	-	Rhodococcus sp.

Turnover Number [1/s]

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

General Information	Comment	Organism
evolution	the enzyme is a member of the crotonase superfamily	Rhodococcus sp.

kcat/KM [mM/s]

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.