Literature summary extracted from

Loschonsky, S.; Waltzer, S.; Fraas, S.; Wacker, T.; Andrade, S.L.; Kroneck, P.M.; Mueller, M.
Catalytic scope of the thiamine-dependent multifunctional enzyme cyclohexane-1,2-dione hydrolase (2014), ChemBioChem, 15, 389-392.

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
2.2.1.6	recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)	Azoarcus sp.
3.7.1.11	recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)	Azoarcus sp.

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
3.7.1.11	cyclohexane-1,2-dione + H2O	Azoarcus sp.	-	6-oxohexanoate	-	?
3.7.1.11	cyclohexane-1,2-dione + H2O	Azoarcus sp. 22Lin	-	6-oxohexanoate	-	?
3.7.1.11	additional information	Azoarcus sp.	the enzyme catalyzes C-C bond cleavage of cyclohexane-1,2-dione to produce 6-oxohexanoic acid as the primary product, presumably followed by oxidation of the latter to adipic acid	?	-	?
3.7.1.11	additional information	Azoarcus sp. 22Lin	the enzyme catalyzes C-C bond cleavage of cyclohexane-1,2-dione to produce 6-oxohexanoic acid as the primary product, presumably followed by oxidation of the latter to adipic acid	?	-	?

Organism

EC Number	Organism	UniProt	Comment	Textmining
2.2.1.6	Azoarcus sp.	-	-	-
2.2.1.6	Azoarcus sp. 22Lin	-	-	-
3.7.1.11	Azoarcus sp.	P0CH62	-	-
3.7.1.11	Azoarcus sp. 22Lin	P0CH62	-	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
2.2.1.6	recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Azoarcus sp.
3.7.1.11	recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Azoarcus sp.

Source Tissue

EC Number	Source Tissue	Comment	Organism	Textmining
3.7.1.11	culture condition:cyclohexane-1,2-diol-grown cell	cyclohexane-1,2-diol as the sole carbon source and electron donor, and nitrate as electron acceptor	Azoarcus sp.	-

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2.2.1.6	1-naphthaldehyde + pyruvate	-	Azoarcus sp.	(1R)-1-hydroxy-1-(naphthalen-1-yl)propan-2-one + CO2	-	?
2.2.1.6	1-naphthaldehyde + pyruvate	-	Azoarcus sp. 22Lin	(1R)-1-hydroxy-1-(naphthalen-1-yl)propan-2-one + CO2	-	?
2.2.1.6	2-bromobenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-bromophenyl)propan-2-one + CO2	-	?
2.2.1.6	2-bromobenzaldehyde + pyruvate	-	Azoarcus sp. 22Lin	(R)-1-hydroxy-1-(2-bromophenyl)propan-2-one + CO2	-	?
2.2.1.6	2-chlorobenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-chlorophenyl)propan-2-one + CO2	-	?
2.2.1.6	2-chlorobenzaldehyde + pyruvate	-	Azoarcus sp. 22Lin	(R)-1-hydroxy-1-(2-chlorophenyl)propan-2-one + CO2	-	?
2.2.1.6	2-fluorobenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-fluorophenyl)propan-2-one + CO2	-	?
2.2.1.6	2-hydroxybenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-hydroxyphenyl)propan-2-one + CO2	-	?
2.2.1.6	2-iodobenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-iodophenyl)propan-2-one + CO2	-	?
2.2.1.6	2-methoxybenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-methoxyphenyl)propan-2-one + CO2	-	?
2.2.1.6	2-methylbenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-methylphenyl)propan-2-one + CO2	-	?
2.2.1.6	2-naphthaldehyde + pyruvate	-	Azoarcus sp.	(1R)-1-hydroxy-1-(naphthalen-2-yl)propan-2-one + CO2	-	?
2.2.1.6	2-nitrobenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(2-nitrophenyl)propan-2-one + CO2	-	?
2.2.1.6	4-(tert-butyl)benzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(4-(tert-butyl)phenyl)propan-2-one + CO2	-	?
2.2.1.6	4-ethoxybenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(4-ethoxyphenyl)propan-2-one + CO2	-	?
2.2.1.6	4-ethylbenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(4-ethylphenyl)propan-2-one + CO2	-	?
2.2.1.6	4-isopropylbenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(4-isopropylphenyl)propan-2-one + CO2	-	?
2.2.1.6	4-phenylbenzaldehyde + pyruvate	-	Azoarcus sp.	(R)-1-hydroxy-1-(4-ethylphenyl)propan-2-one + CO2	-	?
2.2.1.6	additional information	in addition to its physiological C-C bond-cleavage activity, CDH catalyzes the asymmetric cross-benzoin reaction of a broad variety of aromatic aldehydes and pyruvate. In the case of the sterically demanding 4-(tert-butyl)benzaldehyde and 2-naphthaldehyde, the respective 2-hydroxyketone products are obtained in high yield. The recombinant CDH shows the same C-C bond-cleavage and C-C bond-formation activity as the enzyme purified from its native source, Azoarcus sp. strain 22Lin. Enzyme CDH catalyzes the asymmetric cross-benzoin reaction of aromatic aldehydes and (decarboxylated) pyruvate (up to quantitative conversion, 92-99% ee). The enzyme accepts also hydroxybenzaldehydes and nitrobenzaldehydes. On a semipreparative scale, sterically demanding 4-(tert-butyl)benzaldehyde and 2-naphthaldehyde are transformed into the corresponding 2-hydroxy ketone products in high yields, enzyme substrate specificity and enantioselectivity, overview	Azoarcus sp.	?	-	?
2.2.1.6	additional information	in addition to its physiological C-C bond-cleavage activity, CDH catalyzes the asymmetric cross-benzoin reaction of a broad variety of aromatic aldehydes and pyruvate. In the case of the sterically demanding 4-(tert-butyl)benzaldehyde and 2-naphthaldehyde, the respective 2-hydroxyketone products are obtained in high yield. The recombinant CDH shows the same C-C bond-cleavage and C-C bond-formation activity as the enzyme purified from its native source, Azoarcus sp. strain 22Lin. Enzyme CDH catalyzes the asymmetric cross-benzoin reaction of aromatic aldehydes and (decarboxylated) pyruvate (up to quantitative conversion, 92-99% ee). The enzyme accepts also hydroxybenzaldehydes and nitrobenzaldehydes. On a semipreparative scale, sterically demanding 4-(tert-butyl)benzaldehyde and 2-naphthaldehyde are transformed into the corresponding 2-hydroxy ketone products in high yields, enzyme substrate specificity and enantioselectivity, overview	Azoarcus sp. 22Lin	?	-	?
2.2.1.6	pyruvate + benzaldehyde	The enzyme is able to catalyze nonphysiological asymmetric C-C bond formation, the cross-benzoin reaction of benzaldehyde and pyruvate (after decarboxylation) to result in the R-configured 1-hydroxy-1-phenylpropan-2-one (98% ee)	Azoarcus sp.	(R)-phenylacetylcarbinol + CO2	i.e. (R)-1-hydroxy-1-phenylpropan-2-one	?
2.2.1.6	pyruvate + benzaldehyde	The enzyme is able to catalyze nonphysiological asymmetric C-C bond formation, the cross-benzoin reaction of benzaldehyde and pyruvate (after decarboxylation) to result in the R-configured 1-hydroxy-1-phenylpropan-2-one (98% ee)	Azoarcus sp. 22Lin	(R)-phenylacetylcarbinol + CO2	i.e. (R)-1-hydroxy-1-phenylpropan-2-one	?
3.7.1.11	cyclohexane-1,2-dione + H2O	-	Azoarcus sp.	6-oxohexanoate	-	?
3.7.1.11	cyclohexane-1,2-dione + H2O	-	Azoarcus sp. 22Lin	6-oxohexanoate	-	?
3.7.1.11	additional information	the enzyme catalyzes C-C bond cleavage of cyclohexane-1,2-dione to produce 6-oxohexanoic acid as the primary product, presumably followed by oxidation of the latter to adipic acid	Azoarcus sp.	?	-	?
3.7.1.11	additional information	determination of the catalytic activity of recombinant enzyme CDH, both cleavage of cyclohexane-1,2-dione and the cross-benzoin reaction of benzaldehyde and pyruvate. The enzyme is able to catalyze nonphysiological asymmetric C-C bond formation, the cross-benzoin reaction of benzaldehyde and pyruvate (after decarboxylation) to result in the R-configured 1-hydroxy-1-phenylpropan-2-one (98% ee) is performed on an analytical scale. The recombinant CDH shows the same C-C bond-cleavage and C-C bond-formation activity as the enzyme purified from its native source, Azoarcus sp. strain 22Lin. Enzyme CDH catalyzes the asymmetric cross-benzoin reaction of aromatic aldehydes and (decarboxylated) pyruvate (up to quantitative conversion, 92-99% ee). The enzyme accepts also hydroxybenzaldehydes and nitrobenzaldehydes. On a semipreparative scale, sterically demanding 4-(tert-butyl)benzaldehyde and 2-naphthaldehyde are transformed into the corresponding 2-hydroxy ketone products in high yields	Azoarcus sp.	?	-	?
3.7.1.11	additional information	the enzyme catalyzes C-C bond cleavage of cyclohexane-1,2-dione to produce 6-oxohexanoic acid as the primary product, presumably followed by oxidation of the latter to adipic acid	Azoarcus sp. 22Lin	?	-	?
3.7.1.11	additional information	determination of the catalytic activity of recombinant enzyme CDH, both cleavage of cyclohexane-1,2-dione and the cross-benzoin reaction of benzaldehyde and pyruvate. The enzyme is able to catalyze nonphysiological asymmetric C-C bond formation, the cross-benzoin reaction of benzaldehyde and pyruvate (after decarboxylation) to result in the R-configured 1-hydroxy-1-phenylpropan-2-one (98% ee) is performed on an analytical scale. The recombinant CDH shows the same C-C bond-cleavage and C-C bond-formation activity as the enzyme purified from its native source, Azoarcus sp. strain 22Lin. Enzyme CDH catalyzes the asymmetric cross-benzoin reaction of aromatic aldehydes and (decarboxylated) pyruvate (up to quantitative conversion, 92-99% ee). The enzyme accepts also hydroxybenzaldehydes and nitrobenzaldehydes. On a semipreparative scale, sterically demanding 4-(tert-butyl)benzaldehyde and 2-naphthaldehyde are transformed into the corresponding 2-hydroxy ketone products in high yields	Azoarcus sp. 22Lin	?	-	?

Subunits

EC Number	Subunits	Comment	Organism
2.2.1.6	homodimer	the enzyme is a homodimer in solution, the crystal structure shows a homotetramer with one noncovalently bound FAD and one thiamine diphosphate per monomer	Azoarcus sp.
2.2.1.6	homotetramer	in the crystallized form	Azoarcus sp.
3.7.1.11	homodimer	the enzyme is a homodimer in solution, the crystal structure shows a homotetramer with one noncovalently bound FAD and one thiamine diphosphate per monomer	Azoarcus sp.

Synonyms

EC Number	Synonyms	Comment	Organism
3.7.1.11	Cdh	-	Azoarcus sp.

Temperature Optimum [°C]

EC Number	Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
2.2.1.6	30	-	assay at	Azoarcus sp.

pH Optimum

EC Number	pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
2.2.1.6	6.5	-	assay at	Azoarcus sp.

Cofactor

EC Number	Cofactor	Comment	Organism
2.2.1.6	FAD	noncovalently bound, one FAD per monomer	Azoarcus sp.
2.2.1.6	additional information	the crystal structure shows a homotetramer with one noncovalently bound FAD and one thiamine diphosphate per monomer	Azoarcus sp.
2.2.1.6	thiamine diphosphate	dependent on, one thiamine diphosphate per monomer	Azoarcus sp.
3.7.1.11	FAD	noncovalently bound, one FAD per monomer	Azoarcus sp.
3.7.1.11	additional information	the crystal structure shows a homotetramer with one noncovalently bound FAD and one thiamine diphosphate per monomer	Azoarcus sp.
3.7.1.11	thiamine diphosphate	dependent on, one thiamine diphosphate per monomer	Azoarcus sp.

General Information

EC Number	General Information	Comment	Organism
3.7.1.11	physiological function	thiamine diphosphate-dependent cyclohexane-1,2-dione hydrolase is the key enzyme of an anaerobic degradation pathway of alicyclic alcohols	Azoarcus sp.