Literature summary for 4.2.1.34 extracted from

Kronen, M.; Berg, I.A.
Mesaconase/Fumarase FumD in Escherichia coli O157 H7 and promiscuity of Escherichia coli class I fumarases FumA and FumB (2015), PLoS ONE, 10, e0145098 .

Activating Compound

Activating Compound	Comment	Organism	Structure
thiol	aerobic purification results in inactive protein, which can only partly be reactivated with Fe2+ and thiol	Escherichia coli

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli lysY	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.094	-	fumarate	pH 6.9, 37°C	Escherichia coli
0.1	-	mesaconate	pH 6.9, 37°C	Escherichia coli
0.15	-	mesaconate	pH 6.9, 37°C	Escherichia coli
0.2	-	(S)-malate	pH 6.9, 37°C	Escherichia coli
0.21	-	fumarate	pH 6.9, 37°C	Escherichia coli
0.22	-	mesaconate	pH 6.9, 37°C	Escherichia coli
0.4	-	(S)-malate	pH 6.9, 37°C	Escherichia coli
0.78	-	(S)-malate	pH 6.9, 37°C	Escherichia coli
0.8	-	(2S,3S)-tartrate	pH 6.9, 37°C	Escherichia coli
0.9	-	fumarate	pH 6.9, 37°C	Escherichia coli
0.92	-	(S)-citramalate	pH 6.9, 37°C	Escherichia coli
1.04	-	(S)-citramalate	pH 6.9, 37°C	Escherichia coli
1.08	-	(S)-citramalate	pH 6.9, 37°C	Escherichia coli
2	-	(2S,3S)-tartrate	pH 6.9, 37°C	Escherichia coli
2.6	-	(2S,3S)-tartrate	pH 6.9, 37°C	Escherichia coli

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	aerobic purification results in inactive protein, which can only partly be reactivated with Fe2+ and thiol	Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
60000	-	SDS-PAGE	Escherichia coli
60105	-	calculated from sequence	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
mesaconate + H2O	Escherichia coli	-	(S)-citramalate	-	r
mesaconate + H2O	Escherichia coli ATCC 700728 / O157:H7	-	(S)-citramalate	-	r

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-
Escherichia coli	P0AC33	-	-
Escherichia coli	P14407	-	-
Escherichia coli ATCC 700728 / O157:H7	-	-	-
Escherichia coli W3110 / K-12	P0AC33	-	-
Escherichia coli W3110 / K-12	P14407	-	-

Oxidation Stability

Oxidation Stability	Organism
aerobic purification results in inactive protein, which can only partly be reactivated with Fe2+ and thiol	Escherichia coli

Purification (Commentary)

Purification (Comment)	Organism
-	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
(2S,3S)-tartrate	-	Escherichia coli	oxaloacetate + H2O	-	?
(2S,3S)-tartrate	-	Escherichia coli ATCC 700728 / O157:H7	oxaloacetate + H2O	-	?
(2S,3S)-tartrate	-	Escherichia coli W3110 / K-12	oxaloacetate + H2O	-	?
(S)-citramalate	-	Escherichia coli	mesaconate + H2O	-	r
(S)-citramalate	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli	mesaconate + H2O	-	r
(S)-citramalate	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli W3110 / K-12	mesaconate + H2O	-	r
(S)-citramalate	-	Escherichia coli W3110 / K-12	mesaconate + H2O	-	r
(S)-malate	-	Escherichia coli	fumarate + H2O	-	r
(S)-malate	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli	fumarate + H2O	-	r
(S)-malate	-	Escherichia coli ATCC 700728 / O157:H7	fumarate + H2O	-	r
(S)-malate	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli W3110 / K-12	fumarate + H2O	-	r
(S)-malate	-	Escherichia coli W3110 / K-12	fumarate + H2O	-	r
fumarate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3-fold preference for mesaconate over fumarate	Escherichia coli	(S)-malate	-	r
fumarate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3fold preference for mesaconate over fumarate	Escherichia coli	(S)-malate	-	r
fumarate + H2O	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli	(S)-malate	-	r
fumarate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3-fold preference for mesaconate over fumarate	Escherichia coli ATCC 700728 / O157:H7	(S)-malate	-	r
fumarate + H2O	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli W3110 / K-12	(S)-malate	-	r
fumarate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3fold preference for mesaconate over fumarate	Escherichia coli W3110 / K-12	(S)-malate	-	r
mesaconate + H2O	-	Escherichia coli	(S)-citramalate	-	r
mesaconate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3-fold preference for mesaconate over fumarate	Escherichia coli	(S)-citramalate	-	r
mesaconate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3fold preference for mesaconate over fumarate	Escherichia coli	(S)-citramalate	-	r
mesaconate + H2O	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli	(S)-citramalate	-	r
mesaconate + H2O	-	Escherichia coli ATCC 700728 / O157:H7	(S)-citramalate	-	r
mesaconate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3-fold preference for mesaconate over fumarate	Escherichia coli ATCC 700728 / O157:H7	(S)-citramalate	-	r
mesaconate + H2O	the catalytic efficiency of FumA with (S)-citramalate/mesaconate is about 4% of that with fumarate or (S)-malate	Escherichia coli W3110 / K-12	(S)-citramalate	-	r
mesaconate + H2O	heterologously produced FumD is a promiscuous mesaconase/fumarase with a 2- to 3fold preference for mesaconate over fumarate	Escherichia coli W3110 / K-12	(S)-citramalate	-	r

Synonyms

Synonyms	Comment	Organism
FumA	-	Escherichia coli
fumarase A	-	Escherichia coli
FumD	-	Escherichia coli
mesaconase/fumarase	-	Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
0.09437	-	assay at	Escherichia coli
37	-	assay at	Escherichia coli

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.9	-	assay at	Escherichia coli

General Information

General Information	Comment	Organism
metabolism	the enzyme participates in the methylaspartate pathway of glutamate fermentation as well as in the metabolism of various C5-dicarboxylic acids such as mesaconate or L-threo-beta-methylmalate. fumD is clustered with the key genes for two enzymes of the methylaspartate pathway of glutamate fermentation, glutamate mutase and methylaspartate ammonia lyase, converting glutamate to mesaconate	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.05	-	(2S,3S)-tartrate	pH 6.9, 37°C	Escherichia coli
0.635	-	(2S,3S)-tartrate	pH 6.9, 37°C	Escherichia coli
2.13	-	(2S,3S)-tartrate	pH 6.9, 37°C	Escherichia coli
35	-	(S)-citramalate	pH 6.9, 37°C	Escherichia coli
38	-	(S)-citramalate	pH 6.9, 37°C	Escherichia coli
110	-	(S)-citramalate	pH 6.9, 37°C	Escherichia coli
250	-	mesaconate	pH 6.9, 37°C	Escherichia coli
260	-	(S)-malate	pH 6.9, 37°C	Escherichia coli
370	-	(S)-malate	pH 6.9, 37°C	Escherichia coli
580	-	mesaconate	pH 6.9, 37°C	Escherichia coli
750	-	fumarate	pH 6.9, 37°C	Escherichia coli
870	-	(S)-malate	pH 6.9, 37°C	Escherichia coli
2000	-	mesaconate	pH 6.9, 37°C	Escherichia coli
3120	-	fumarate	pH 6.9, 37°C	Escherichia coli
6600	-	fumarate	pH 6.9, 37°C	Escherichia coli