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Literature summary for 2.5.1.6 extracted from

  • Niu, W.; Cao, S.; Yang, M.; Xu, L.
    Enzymatic synthesis of S-adenosylmethionine using immobilized methionine adenosyltransferase variants on the 50-mM scale (2017), Catalysts, 7, 238 .
No PubMed abstract available
Search for more literature for 2.5.1.6

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli DH5alpha Escherichia coli

Protein Variants

Protein Variants Comment Organism
I303V the Km-values for both substrates are slightly less than those of the wild-type enzyme. The variant is successfully produced at a high level (about 800 mg/l) with approximately four-fold higher specific activity than the wild-type enzyme. The recombinant mutant enzyme is covalently immobilized onto the amino resin and epoxy resin in order to obtain a robust biocatalyst to be used in industrial bioreactors. The immobilized preparation using amino resin exhibits the highest activity coupling yield (about 84%), compared with approximately 3% for epoxy resin. The immobilized mutant enzyme is more stable than the soluble enzyme under the reactive conditions, with a half-life of 229.5 h at 37 °C. The Km(ATP) value (0.18 mM) of the immobilized mutant enzyme is about two-fold lower than that of the soluble enzyme. The immobilized enzyme shows high operational stability during 10 consecutive 8 h batches, with the substrate adenosine triphosphate conversion rate above 95% on the 50 mM scale. Compared with the wild-type enzyme, as little as 200 mM sodium p-toluenesulfonate is required to completely overcome the product inhibition by S-adenosyl-L-methionine of I303V mutant enzyme on a 30 mM scale incubation Escherichia coli

General Stability

General Stability Organism
a wider range of pH stability is obtained with the immobilized enzym Escherichia coli
the immobilized enzyme shows high operational stability during 10 consecutive 8 h batches Escherichia coli
thermal stability of the immobilized enzyme is much higher than that of the soluble enzyme Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
S-adenosyl-L-methionine product inhibition. Compared with the wild-type MAT, as little as 200 mM sodium p-toluenesulfonate is required to completely overcome the product inhibition of I303V mutant enzyme on a 30 mM scale incubation Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.12
-
 L-methionine 37°C, pH 8.0, mutant enzyme I303V Escherichia coli
0.18
-
 L-methionine 37°C, pH 8.0, wild-type enzyme Escherichia coli
0.34
-
 ATP 37°C, pH 8.0, mutant enzyme I303V Escherichia coli
0.43
-
 ATP 37°C, pH 8.0, wild-type enzyme Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-methionine + H2O Escherichia coli
-
 phosphate + diphosphate + S-adenosyl-L-methionine
-
 ?
ATP + L-methionine + H2O Escherichia coli BL21 (DE3)
-
 phosphate + diphosphate + S-adenosyl-L-methionine
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Escherichia coli BL21 (DE3)
-
-
-

Purification (Commentary)

Purification (Comment) Organism
His6-tagged wild-type and I303V mutant proteins are purified in a single chromatography step usinf a HiTrap chelating nickel column Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-methionine + H2O
-
 Escherichia coli phosphate + diphosphate + S-adenosyl-L-methionine
-
 ?
ATP + L-methionine + H2O
-
 Escherichia coli BL21 (DE3) phosphate + diphosphate + S-adenosyl-L-methionine
-
 ?

Synonyms

Synonyms Comment Organism
MAT
-
 Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
 30% loss of activity after 12 h, complete loss of activity after 36-48 h, soluble enzyme. 63% of the initial enzyme activity of immobilized MAT was remained after 168 h incubation. Comparing the t1/2 values at 37 °C, the immobilized enzyme is almost 25fold more stable than the free enzyme Escherichia coli

pH Stability

pH Stability pH Stability Maximum Comment Organism
6.5 8.5 4°C, 12 h, immobilized enzyme remains stable Escherichia coli
7 8 4°C, 12 h, soluble enzyme remains stable Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.02
-
 S-adenosyl-L-methionine 37°C, pH 8.0, wild-type enzyme Escherichia coli
0.12
-
 S-adenosyl-L-methionine 37°C, pH 8.0, mutant enzyme I303V Escherichia coli