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Literature summary for 2.3.1.46 extracted from

  • Ziegler, K.; Noble, S.M.; Mutumanje, E.; Bishop, B.; Huddler, D.P.; Born, T.L.
    Identification of catalytic cysteine, histidine, and lysine residues in Escherichia coli homoserine transsuccinylase (2007), Biochemistry, 46, 2674-2683.
    View publication on PubMed
Search for more literature for 2.3.1.46

Cloned(Commentary)

Cloned (Comment) Organism
gene metA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
C142A site-directed mutagenesis, the mutant is inactive Escherichia coli
C142S site-directed mutagenesis, the mutant is inactive Escherichia coli
C90A site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Escherichia coli
C90S site-directed mutagenesis, the mutant shows only slightly reduced activity compared to the wild-type enzyme Escherichia coli
H235A site-directed mutagenesis, the mutant is inactive Escherichia coli
H235N site-directed mutagenesis, the mutant is inactive Escherichia coli
K45A/K46A site-directed mutagenesis, the mutant is nearly inactive Escherichia coli
K46A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
K46R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
K46R/K47R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
K47A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Escherichia coli
K47R site-directed mutagenesis, the mutant shows 90% reduced activity compared to the wild-type enzyme Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
diethyldicarbonate inactivation, pH dependence, overview Escherichia coli
iodoacetamide inactivation, pH dependence, overview Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information steady-state kinetics Escherichia coli
0.0032
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
0.027
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
0.044
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
0.049
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
0.094
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
0.13
-
 succinyl-CoA pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
0.13
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
0.13
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
0.14
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant C90S Escherichia coli
0.14
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
0.16
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
0.58
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
0.72
-
 L-homoserine pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
1.07
-
 L-homoserine pH 7.2, 25°C, recombinant mutant C90S Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
succinyl-CoA + L-homoserine Escherichia coli step in the biosynthetic pathways of methionine and S-adenosylmethionine, overview CoA + O-succinyl-L-homoserine
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
 gene metA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange chromatography, ammonium sulfate fractionation, hydrophobic interaction chromatography, and dialysis to over 90% purity Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
succinyl-CoA + L-homoserine = CoA + O-succinyl-L-homoserine ping pong catalytic reaction mechanism, the residues Cys142, His235, and Lys47 are essential for catalytic activity, Cys142 acts as nucleophile to which the succinyl moiety is transferred during the reaction followed by transfer to homoserine to form O-succinylhomoserine, Lys47 is involved in the first half-reaction, and His235 is the active site base Escherichia coli
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
succinyl-CoA + L-homoserine
-
 Escherichia coli CoA + O-succinyl-L-homoserine
-
 ?
succinyl-CoA + L-homoserine step in the biosynthetic pathways of methionine and S-adenosylmethionine, overview Escherichia coli CoA + O-succinyl-L-homoserine
-
 ?

Synonyms

Synonyms Comment Organism
homoserine transsuccinylase
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.034
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
0.34
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47A Escherichia coli
1.8
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
1.8
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K47R Escherichia coli
2
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
2
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R/K47R Escherichia coli
4.1
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
4.1
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46A Escherichia coli
37
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
37
-
 L-homoserine pH 7.2, 25°C, recombinant mutant K46R Escherichia coli
130
-
 succinyl-CoA pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
130
-
 L-homoserine pH 7.2, 25°C, recombinant wild-type enzyme Escherichia coli
130
-
 succinyl-CoA pH 7.2, 25°C, recombinant mutant C90S Escherichia coli
130
-
 L-homoserine pH 7.2, 25°C, recombinant mutant C90S Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
 assay at Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
6.2 8.5 pH profile of mutant K47R Escherichia coli