Literature summary for 1.3.1.32 extracted from

Chen, L.; Krol, E.S.; Sakharkar, M.K.; Khan, H.A.; Alhomida, A.S.; Yang, J.
Residues His172 and Lys238 are essential for the catalytic activity of the maleylacetate reductase from Sphingobium chlorophenolicum strain L-1 (2017), Sci. Rep., 7, 18097 .

Search for more literature for 1.3.1.32

Application

Application	Comment	Organism
environmental protection	further evolution to more catalytically active PcpE may be an important contributor to improved Sphingobium chlorophenolicum L-1-mediated bioremediation of pentachlorophenol	Sphingobium chlorophenolicum

Cloned(Commentary)

Cloned (Comment)	Organism
-	Sphingobium chlorophenolicum

Protein Variants

Protein Variants	Comment	Organism
H172A	no catalytic activity	Sphingobium chlorophenolicum
H172A	no catalytic activity detected	Sphingobium chlorophenolicum
H236A	kcat/Km for 2-maleylacetate is 2.3fold higher as compared to wild-type value	Sphingobium chlorophenolicum
H236A	mutation increases the catalytic efficiency (kcat/Km) of the enzyme (PcpE) by more than 2fold	Sphingobium chlorophenolicum
H237A	kcat/Km for 2-maleylacetate is 3.2fold lower as compared to wild-type value	Sphingobium chlorophenolicum
H237A	Km and kcat are augmented with a reduction in the catalytic efficiency kcat/Km	Sphingobium chlorophenolicum
H241A	kcat/Km for 2-maleylacetate is 1.2fold lower as compared to wild-type value	Sphingobium chlorophenolicum
H241A	Km and kcat are augmented with a reduction in the catalytic efficiency kcat/Km	Sphingobium chlorophenolicum
H251A	kcat/Km for 2-maleylacetate is 2.6fold lower as compared to wild-type value	Sphingobium chlorophenolicum
H251A	Km and kcat are augmented with a reduction in the catalytic efficiency kcat/Km	Sphingobium chlorophenolicum
K140A	kcat/Km for 2-maleylacetate is 5.3fold lower as compared to wild-type value	Sphingobium chlorophenolicum
K140A	the mutant exhibits a 4fold increase in Km and a slight decrease in kcat compared to the wild type enzyme, resulting in a 5.3-fold decrease of the catalytic efficiency kcat/Km	Sphingobium chlorophenolicum
K238A	no catalytic activity	Sphingobium chlorophenolicum
K238A	no catalytic activity detected	Sphingobium chlorophenolicum

Inhibitors

Inhibitors	Comment	Organism	Structure
Pentachlorophenol	inhibits in a concentration-dependent manner	Sphingobium chlorophenolicum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.07	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H236A, cosubstrate: NADH	Sphingobium chlorophenolicum
0.09	-	2-chloromaleylacetate	pH 7.0, 23°C	Sphingobium chlorophenolicum
0.09	-	2-maleylacetate	pH 7.0, 23°C	Sphingobium chlorophenolicum
0.09	-	2-chloromaleylacetate	pH 7.0, 23°C, wild-type enzyme, cosubstrate: NADH	Sphingobium chlorophenolicum
0.09	-	2-maleylacetate	pH 7.0, 23°C, wild-type enzyme, cosubstrate: NADH	Sphingobium chlorophenolicum
0.13	-	NADH	cosubstrate: 2-chloromaleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
0.13	-	NADH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-chloromaleylacetate	Sphingobium chlorophenolicum
0.16	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H241A, cosubstrate: NADH	Sphingobium chlorophenolicum
0.36	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme K140A, cosubstrate: NADH	Sphingobium chlorophenolicum
0.44	-	NADH	cosubstrate: 2-maleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
0.44	-	NADH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-maleylacetate	Sphingobium chlorophenolicum
0.52	-	NADPH	cosubstrate: 2-maleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
0.52	-	NADPH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-maleylacetate	Sphingobium chlorophenolicum
0.57	-	NADPH	cosubstrate: 2-chloromaleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
0.67	-	NADPH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-chloromaleylacetate	Sphingobium chlorophenolicum
0.93	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H237A, cosubstrate: NADH	Sphingobium chlorophenolicum
1.69	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H251A, cosubstrate: NADH	Sphingobium chlorophenolicum

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2-chloromaleylacetate + NADH + H+	Sphingobium chlorophenolicum	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	maleylacetate + NAD+ + HCl	-	?
2-chloromaleylacetate + NADH + H+	Sphingobium chlorophenolicum	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	maleylacetate + NAD+ + Cl-	-	?
2-chloromaleylacetate + NADH + H+	Sphingobium chlorophenolicum L-1	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	maleylacetate + NAD+ + Cl-	-	?
2-chloromaleylacetate + NADH + H+	Sphingobium chlorophenolicum L-1	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	maleylacetate + NAD+ + HCl	-	?
2-maleylacetate + NADH + H+	Sphingobium chlorophenolicum	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	Sphingobium chlorophenolicum	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	Sphingobium chlorophenolicum L-1	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	Sphingobium chlorophenolicum L-1	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	3-oxoadipate + NAD+	-	?

Organism

Organism	UniProt	Comment	Textmining
Sphingobium chlorophenolicum	-	-	-
Sphingobium chlorophenolicum	Q8KN40	-	-
Sphingobium chlorophenolicum L-1	-	-	-
Sphingobium chlorophenolicum L-1	Q8KN40	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Sphingobium chlorophenolicum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2-chloromaleylacetate + NADH + H+	-	Sphingobium chlorophenolicum	maleylacetate + NAD+ + HCl	-	?
2-chloromaleylacetate + NADH + H+	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum	maleylacetate + NAD+ + HCl	-	?
2-chloromaleylacetate + NADH + H+	-	Sphingobium chlorophenolicum L-1	maleylacetate + NAD+ + HCl	-	?
2-chloromaleylacetate + NADH + H+	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum L-1	maleylacetate + NAD+ + HCl	-	?
2-chloromaleylacetate + NADH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum	maleylacetate + NAD+ + Cl-	-	?
2-chloromaleylacetate + NADH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum L-1	maleylacetate + NAD+ + Cl-	-	?
2-chloromaleylacetate + NADPH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum	maleylacetate + NADP+ + Cl-	-	?
2-chloromaleylacetate + NADPH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum L-1	maleylacetate + NADP+ + Cl-	-	?
2-chloromaleylacetate + NADPH + H+	-	Sphingobium chlorophenolicum	maleylacetate + NADP+ + HCl	-	?
2-maleylacetate + NADH + H+	-	Sphingobium chlorophenolicum	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum L-1	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	-	Sphingobium chlorophenolicum L-1	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADH + H+	maleylacetate reductase (PcpE) is the last enzyme in the pentachlorophenol biodegradation pathway in Sphingobium chlorophenolicum L-1. It catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum L-1	3-oxoadipate + NAD+	-	?
2-maleylacetate + NADPH + H+	-	Sphingobium chlorophenolicum	3-oxoadipate + NADP+	-	?
2-maleylacetate + NADPH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum	3-oxoadipate + NADP+	-	?
2-maleylacetate + NADPH + H+	the enzyme catalyzes two consecutive reductive reactions, reductive dehalogenation of 2-chloromaleylacetate to maleylacetate and subsequent reduction of maleylacetate to 3-oxoadipate	Sphingobium chlorophenolicum L-1	3-oxoadipate + NADP+	-	?
2-maleylacetate + NADPH + H+	-	Sphingobium chlorophenolicum L-1	3-oxoadipate + NADP+	-	?

Synonyms

Synonyms	Comment	Organism
maleylacetate reductase	-	Sphingobium chlorophenolicum
PcpE	-	Sphingobium chlorophenolicum

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.7	-	NADPH	cosubstrate: 2-maleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
0.7	-	NADPH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-maleylacetate	Sphingobium chlorophenolicum
0.9	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme K140A, cosubstrate: NADH	Sphingobium chlorophenolicum
1.2	-	2-maleylacetate	pH 7.0, 23°C	Sphingobium chlorophenolicum
1.2	-	NADH	cosubstrate: 2-maleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
1.2	-	NADH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-maleylacetate	Sphingobium chlorophenolicum
1.2	-	2-maleylacetate	pH 7.0, 23°C, wild-type enzyme, cosubstrate: NADH	Sphingobium chlorophenolicum
1.7	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H241A, cosubstrate: NADH	Sphingobium chlorophenolicum
2.1	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H236A, cosubstrate: NADH	Sphingobium chlorophenolicum
3.8	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H237A, cosubstrate: NADH	Sphingobium chlorophenolicum
8.8	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H251A, cosubstrate: NADH	Sphingobium chlorophenolicum
17.6	-	2-chloromaleylacetate	pH 7.0, 23°C	Sphingobium chlorophenolicum
17.6	-	2-chloromaleylacetate	pH 7.0, 23°C, wild-type enzyme, cosubstrate: NADH	Sphingobium chlorophenolicum
29.7	-	NADPH	cosubstrate: 2-chloromaleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
29.7	-	NADPH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-chloromaleylacetate	Sphingobium chlorophenolicum
143.7	-	NADH	cosubstrate: 2-chloromaleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
143.7	-	NADH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-chloromaleylacetate	Sphingobium chlorophenolicum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	-	Sphingobium chlorophenolicum
7	-	maximal NADH-dependent catalytic activity, substrate: 2-chloromaleylacetate	Sphingobium chlorophenolicum

Cofactor

Cofactor	Comment	Organism	Structure
NADH	the enzyme preferentially binds NADH to NADPH	Sphingobium chlorophenolicum
NADH	the NADH/NADPH substrate specificity (kcat/Km ratio) is 25.0 with 2-chloromaleylacetate as cosubstrate and 2.1 with 2-maleylacetate as cosubstrate	Sphingobium chlorophenolicum
NADPH	the enzyme preferentially binds NADH to NADPH	Sphingobium chlorophenolicum
NADPH	the NADH/NADPH substrate specificity (kcat/Km ratio) is 25.0 with 2-chloromaleylacetate as cosubstrate and 2.1 with 2-maleylacetate as cosubstrate	Sphingobium chlorophenolicum

General Information

General Information	Comment	Organism
evolution	mutagenesis studies show that His172 and Lys238 are essential for the catalytic activity of the enzyme (PcpE). However,the mutation of His236 to an alanine can increase the catalytic efficiency (kcat/Km) of PcpE by more than 2fold, implying that PcpE is still in an early stage of molecular evolution. The enzyme (PcpE) exhibits an extremely low but detectable level of alcohol dehalogenase activity toward ethanol and supports the notion that it is evolved from an iron-containing alcohol dehydrogenase	Sphingobium chlorophenolicum
metabolism	last enzyme in the pentachlorophenol biodegradation pathway	Sphingobium chlorophenolicum

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
1.3	-	NADPH	cosubstrate: 2-maleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
1.3	-	NADPH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-maleylacetate	Sphingobium chlorophenolicum
2.5	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme K140A, cosubstrate: NADH	Sphingobium chlorophenolicum
2.7	-	NADH	cosubstrate: 2-maleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
2.7	-	NADH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-maleylacetate	Sphingobium chlorophenolicum
4.1	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H237A, cosubstrate: NADH	Sphingobium chlorophenolicum
5.2	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H251A, cosubstrate: NADH	Sphingobium chlorophenolicum
10.6	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H241A, cosubstrate: NADH	Sphingobium chlorophenolicum
13.3	-	2-maleylacetate	pH 7.0, 23°C, wild-type enzyme, cosubstrate: NADH	Sphingobium chlorophenolicum
30	-	2-maleylacetate	pH 7.0, 23°C, mutant enzyme H236A, cosubstrate: NADH	Sphingobium chlorophenolicum
44.3	-	NADPH	cosubstrate: 2-chloromaleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
44.3	-	NADPH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-chloromaleylacetate	Sphingobium chlorophenolicum
195.6	-	2-chloromaleylacetate	pH 7.0, 23°C	Sphingobium chlorophenolicum
195.6	-	2-chloromaleylacetate	pH 7.0, 23°C, wild-type enzyme, cosubstrate: NADH	Sphingobium chlorophenolicum
1105.4	-	NADH	cosubstrate: 2-chloromaleylacetate, pH 7.0, 23°C	Sphingobium chlorophenolicum
1105.4	-	NADH	pH 7.0, 23°C, wild-type enzyme, cosubstrate: 2-chloromaleylacetate	Sphingobium chlorophenolicum

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Release 2023.1 (January 2023)