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Literature summary extracted from

  • Gokhale, R.S.; Agarwalla, S.; Santi, D.V.; Balaram, P.
    Covalent reinforcement of a fragile region in the dimeric enzyme thymidylate synthase stabilizes the protein against chaotrope-induced unfolding (1996), Biochemistry, 35, 7150-7158.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.45 cloned and overexpressed in Escherichia coli Lacticaseibacillus casei
2.1.1.45 plasmid pSCTS9, synthetic TS gene in pUC18 Lacticaseibacillus casei

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.45
-
Lacticaseibacillus casei

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.45 T155C/E188C/C244T formation of 2 disulfide crosslinks across the subunit interface results in dramatic stabilization of the overall structure of the protein, retention of secondary structure at temperatures as high as 90°C, engineering of an intersubunit crosslink yields a fully active enzyme Lacticaseibacillus casei

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.45 5-fluoro-2'-deoxyuridylic acid
-
Lacticaseibacillus casei

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.45 35000
-
2 * 35000, SDS-PAGE Lacticaseibacillus casei
2.1.1.45 36620
-
calculated molecular mass based on the sequence Lacticaseibacillus casei
2.1.1.45 71000
-
analytical ultracentrifugation Lacticaseibacillus casei

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Lacticaseibacillus casei
-
dihydrofolate + dTMP
-
?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP Tequatrovirus T4
-
dihydrofolate + dTMP
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.45 Lacticaseibacillus casei
-
-
-
2.1.1.45 Tequatrovirus T4
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
Lacticaseibacillus casei dihydrofolate + dTMP
-
?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
Lacticaseibacillus casei dihydrofolate + dTMP
-
r
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
Tequatrovirus T4 dihydrofolate + dTMP
-
?
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
Tequatrovirus T4 dihydrofolate + dTMP
-
r

Subunits

EC Number Subunits Comment Organism
2.1.1.45 dimer
-
Tequatrovirus T4
2.1.1.45 dimer 2 * 35000, SDS-PAGE Lacticaseibacillus casei

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.45 55
-
mutant enzyme, 2 disulfide bridges across the dimer interface of the enzyme engineered Lacticaseibacillus casei

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.1.1.45 30 70 mutant enzyme, 2 disulfide bridges across the dimer interface of the enzyme engineered Lacticaseibacillus casei

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.1.1.45 52
-
wild-type enzyme, precipitates at 52°C Lacticaseibacillus casei
2.1.1.45 90
-
mutant enzyme remains soluble, enhancement of activity is observed up to 60°C, possesses appreciable enzymatic activity at 65°C and retains secondary structure even at 90°C Lacticaseibacillus casei

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.1.1.45 0.000005
-
5-fluoro-2'-deoxyuridylic acid
-
Lacticaseibacillus casei