Literature summary for 1.14.12.3 extracted from

Swift, R.J.; Carter, S.F.; Widdowson, D.A.; Mason, J.R.; Leak, D.J.
Expression of benzene dioxygenase from Pseudomonas putida ML2 in cis-1,2-cyclohexanediol-degrading pseudomonads (2001), Appl. Microbiol. Biotechnol., 55, 721-726.

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Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli JM109	Pseudomonas putida
genes bedC1C2BA, under the control of the tac promoter, subcloned into pLAFR5, successfully conjugated into seven of the Gram-negative cis-1,2-cyclohexanediol-degrading isolates and stably maintained and expressed in three of them, strain CHD1CHD3, the strains grow on cis-1,2-cyclohexanediol as sole carbon source, express an active BDO and oxidise cyclohexene, but none of the three strains is able to grow on cyclohexene as sole carbon source, overview, expression in Escherichia coli strain JM109	Pseudomonas putida

Cloned (Comment)

Organism

expression in Escherichia coli JM109

Pseudomonas putida

genes bedC1C2BA, under the control of the tac promoter, subcloned into pLAFR5, successfully conjugated into seven of the Gram-negative cis-1,2-cyclohexanediol-degrading isolates and stably maintained and expressed in three of them, strain CHD1CHD3, the strains grow on cis-1,2-cyclohexanediol as sole carbon source, express an active BDO and oxidise cyclohexene, but none of the three strains is able to grow on cyclohexene as sole carbon source, overview, expression in Escherichia coli strain JM109

Pseudomonas putida

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	required	Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
1-methylcyclohexene + O2 + NADH	Pseudomonas putida	-	?	-	?
1-methylcyclohexene + O2 + NADH	Pseudomonas putida ML2	-	?	-	?
2 cyclohexene + 2 O2 + 2 NADH + 2 H+	Pseudomonas putida	-	2-cyclohexen-1-ol + cis-1,2-cyclohexanediol + 2 NAD+ + H2O	failure of transformed bacteria to grow on cyclohexene is attributed to the toxicity of metabolic intermediates accumulating from the 2-cyclohexen-1-ol metabolism, cyclohexene metabolism pathways, overview	?
3-methylcyclohexene + O2 + NADH	Pseudomonas putida	-	?	-	?
3-methylcyclohexene + O2 + NADH	Pseudomonas putida ML2	-	?	-	?
benzene + O2 + NADH	Pseudomonas putida	-	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	-	?
benzene + O2 + NADH	Pseudomonas putida ML2	-	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	-	?
toluene + O2 + NADH	Pseudomonas putida	-	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	-	-	-
Pseudomonas putida	-	gene bedC1C2BA	-
Pseudomonas putida ML2	-	-	-
Pseudomonas putida ML2	-	gene bedC1C2BA	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.0049	-	recombinant enzyme expressed in Escherichia coli, substrate cyclohexene	Pseudomonas putida
0.0077	-	recombinant enzyme expressed in Escherichia coli, substrate 1-methylcyclohexene	Pseudomonas putida
0.0086	-	recombinant enzyme expressed in Escherichia coli, substrate 3-methylcyclohexene	Pseudomonas putida
0.0172	-	recombinant enzyme expressed in Escherichia coli, substrate toluene	Pseudomonas putida
0.0236	-	recombinant enzyme expressed in Escherichia coli, substrate benzene	Pseudomonas putida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
1-methylcyclohexene + O2 + NADH	-	Pseudomonas putida	?	-	?
1-methylcyclohexene + O2 + NADH	-	Pseudomonas putida ML2	?	-	?
2 cyclohexene + 2 O2 + 2 NADH + 2 H+	-	Pseudomonas putida	2-cyclohexen-1-ol + cis-1,2-cyclohexanediol + 2 NAD+ + H2O	-	?
2 cyclohexene + 2 O2 + 2 NADH + 2 H+	-	Pseudomonas putida	2-cyclohexen-1-ol + cis-1,2-cyclohexanediol + 2 NAD+ + H2O	failure of transformed bacteria to grow on cyclohexene is attributed to the toxicity of metabolic intermediates accumulating from the 2-cyclohexen-1-ol metabolism, cyclohexene metabolism pathways, overview	?
3-methylcyclohexene + O2 + NADH	-	Pseudomonas putida	?	-	?
3-methylcyclohexene + O2 + NADH	-	Pseudomonas putida ML2	?	-	?
benzene + NADH + O2	-	Pseudomonas putida	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	the enzyme oxidizes cyclohexene to a mixture of two products, a monohydroxylated 2-cyclohexen-1-ol product and a dihydroxylated cis-1,2-cyclohexanediol	?
benzene + NADH + O2	-	Pseudomonas putida ML2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	the enzyme oxidizes cyclohexene to a mixture of two products, a monohydroxylated 2-cyclohexen-1-ol product and a dihydroxylated cis-1,2-cyclohexanediol	?
benzene + O2 + NADH	-	Pseudomonas putida	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	-	?
benzene + O2 + NADH	best substrate	Pseudomonas putida	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	-	?
benzene + O2 + NADH	-	Pseudomonas putida ML2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	-	?
benzene + O2 + NADH	best substrate	Pseudomonas putida ML2	cis-cyclohexa-3,5-diene-1,2-diol + NAD+	-	?
toluene + O2 + NADH	-	Pseudomonas putida	?	-	?

Synonyms

Synonyms	Comment	Organism
BDO	-	Pseudomonas putida
benzene dioxygenase	-	Pseudomonas putida

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Pseudomonas putida

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Pseudomonas putida

Cofactor

Cofactor	Comment	Organism	Structure
NADH	-	Pseudomonas putida