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Literature summary for 2.6.1.52 extracted from

  • Zhang, Y.; Ma, C.; Dischert, W.; Soucaille, P.; Zeng, A.P.
    Engineering of phosphoserine aminotransferase increases the conversion of L-homoserine to 4-hydroxy-2-ketobutyrate in a glycerol-independent pathway of 1,3-propanediol production from glucose (2019), Biotechnol. J., 14, e1900003 .
    View publication on PubMed
Search for more literature for 2.6.1.52

Cloned(Commentary)

Cloned (Comment) Organism
coexpression of enzyme mutant SerCR42W/R77W with Escherichia coli pyruvate decarboxylase and alcohol dehydrogenase in Escherichia coli strain BL21(DE3), recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
H41K site-directed mutagenesis Escherichia coli
H41Q site-directed mutagenesis Escherichia coli
H41Y site-directed mutagenesis Escherichia coli
additional information phosphoserine aminotransferase (SerC) from Escherichia coli strain MG1655 is engineered to catalyze the deamination of homoserine to 4-hydroxy-2-oxobutyrate, a key reaction in producing 1,3-propanediol (1,3-PDO) from glucose in a distinct glycerol-independent metabolic pathway. An computation-based rational approach is used to change the substrate specificity of SerC from L-phosphoserine to L-homoserine, molecular dynamics simulations and virtual screening are combined to predict mutation sites. The coexpression of best mutant SerCR42W/R77W with Escherichia coli pyruvate decarboxylase and alcohol dehydrogenase results in production of 3.03 g/l of 1,3-PDO in fed-batch fermentation, which is 13fold higher than in the wild-type strain. Method evaluation, overview Escherichia coli
R42E site-directed mutagenesis Escherichia coli
R42K site-directed mutagenesis Escherichia coli
R42Q site-directed mutagenesis Escherichia coli
R42W site-directed mutagenesis, the mutant exhibits high activity toward L-homoserine with a low activity toward L-phosphoserine Escherichia coli
R42W/R329G site-directed mutagenesis Escherichia coli
R42W/R329H site-directed mutagenesis Escherichia coli
R42W/R329Q site-directed mutagenesis Escherichia coli
R42W/R77E site-directed mutagenesis Escherichia coli
R42W/R77I site-directed mutagenesis Escherichia coli
R42W/R77S site-directed mutagenesis Escherichia coli
R42W/R77T site-directed mutagenesis Escherichia coli
R42W/R77W site-directed mutagenesis Escherichia coli
R42W/R77W site-directed mutagenesis, best engineered mutant showing 4.2fold increased activity with L-homoserine compared to wild-type enzyme, the activity toward the natural substrate L-phosphoserine is highly reduced in the mutant enzyme Escherichia coli
R42W/R77WH328E site-directed mutagenesis, very low activity with L-homoserine Escherichia coli
R42W/R77WH328K site-directed mutagenesis, no activity with L-homoserine Escherichia coli
R42W/R77WH328L site-directed mutagenesis Escherichia coli
R42W/R77WH328Y site-directed mutagenesis Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
14.15
-
 L-homoserine pH 8.2, 30°C, recombinant mutant R42W/R77W Escherichia coli
158.5
-
 L-homoserine pH 8.2, 30°C, recombinant wild-type enzyme Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
O-phospho-L-serine + 2-oxoglutarate Escherichia coli
-
 3-phosphooxypyruvate + L-glutamate
-
 r
O-phospho-L-serine + 2-oxoglutarate Escherichia coli MG1655
-
 3-phosphooxypyruvate + L-glutamate
-
 r

Organism

Organism UniProt Comment Textmining
Escherichia coli P23721
-
-
Escherichia coli MG1655 P23721
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.007
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77W/H328E Escherichia coli
0.01
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant H41K Escherichia coli
0.022
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42E Escherichia coli
0.031
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant H41Y Escherichia coli
0.036
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77W/H328L Escherichia coli
0.051
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant H41Q Escherichia coli
0.058
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77E Escherichia coli
0.079
-
 substrate L-phosphoserine, pH 8.2, 30°C, recombinant mutant R42W/R77W Escherichia coli
0.08
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77W/H328Y Escherichia coli
0.115
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant wild-type enzyme Escherichia coli
0.131
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42K Escherichia coli
0.145
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R329G Escherichia coli
0.148
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42Q Escherichia coli
0.18
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77I Escherichia coli
0.224
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77S Escherichia coli
0.266
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W Escherichia coli
0.273
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77T Escherichia coli
0.281
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R329Q Escherichia coli
0.293
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R329H Escherichia coli
0.487
-
 substrate L-homoserine, pH 8.2, 30°C, recombinant mutant R42W/R77W Escherichia coli
3.406
-
 pH 8.2, 30°C, substrate L-phosphoserine, recombinant wild-type enzyme Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-homoserine + 2-oxoglutarate L-homoserine transaminase reaction of engineered enzyme mutant R42W/R77W, low activity with the wild-type enzyme Escherichia coli 4-hydroxy-2-oxobutyrate + L-glutamate
-
 r
L-homoserine + 2-oxoglutarate L-homoserine transaminase reaction of engineered enzyme mutant R42W/R77W, low activity with the wild-type enzyme Escherichia coli MG1655 4-hydroxy-2-oxobutyrate + L-glutamate
-
 r
O-phospho-L-serine + 2-oxoglutarate
-
 Escherichia coli 3-phosphooxypyruvate + L-glutamate
-
 r
O-phospho-L-serine + 2-oxoglutarate
-
 Escherichia coli MG1655 3-phosphooxypyruvate + L-glutamate
-
 r

Subunits

Subunits Comment Organism
? x * 39000, recombinant enzyme mutant R42W/R77W, SDS-PAGE Escherichia coli

Synonyms

Synonyms Comment Organism
phosphoserine aminotransferase
-
 Escherichia coli
serC
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate PLP Escherichia coli