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Literature summary for 2.1.1.157 extracted from

  • Waditee, R.; Tanaka, Y.; Aoki, K.; Hibino, T.; Jikuya, H.; Takano, J.; Takabe, T.
    Isolation and functional characterization of N-methyltransferases that catalyze betaine synthesis from glycine in a halotolerant photosynthetic organism Aphanothece halophytica (2003), J. Biol. Chem., 278, 4932-4942.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information abiotic stess-inducible enzyme, e.g. high salt content Aphanothece halophytica

Cloned(Commentary)

Cloned (Comment) Organism
orf2, DNA and amino acid sequence determination and analysis, phylogenetic analysis, overexpression of wild-type and mutant enzymes in Escherichia coli, functional coexpression of the soluble enzyme with the glycine sarcosine methyltransferase, EC 2.1.1.156, increases the intracellular betaine level and the growth rate of Escherichia coli strain BL21(DE3) Aphanothece halophytica

Protein Variants

Protein Variants Comment Organism
M172R site-directed mutagenesis, reduced activity with sarcosine, but highly reduced activity with dimethylglycine compared to the wild-type enzyme Aphanothece halophytica
P171Q site-directed mutagenesis, reduced activity with sarcosine and dimethylglycine compared to the wild-type enzyme Aphanothece halophytica
P171Q/M172R site-directed mutagenesis, reduced activity with glycine and sarcosine, but highly reduced activity with dimethylglycine compared to the wild-type enzyme Aphanothece halophytica

Inhibitors

Inhibitors Comment Organism Structure
dimethylglycine competitive inhibition, versus sarcosine, 40% inhibition at 250 mM Aphanothece halophytica
isovaleric acid 10% inhibition at 250 mM, with dimethylglycine Aphanothece halophytica
L-alanine 10% inhibition at 250 mM, with dimethylglycine Aphanothece halophytica
L-cysteine 10% inhibition at 250 mM, with dimethylglycine Aphanothece halophytica
L-methionine 10% inhibition at 250 mM, with dimethylglycine Aphanothece halophytica
L-phenylalanine 10% inhibition at 250 mM Aphanothece halophytica
additional information inhibition rates with sarcosine, overview, no inhibition with dimethylglycine by glycine, acetate, ethanolamine, monomethylethanolamine, serine, asparagine, glutamate, and proline Aphanothece halophytica
n-butyric acid 50% inhibition at 250 mM, with dimethylglycine Aphanothece halophytica
propionic acid 10% inhibition at 250 mM, with dimethylglycine Aphanothece halophytica
S-adenosyl-L-homocysteine product inhibition Aphanothece halophytica
sarcosine slight competitive inhibition, versus dimethylglycine, 20% inhibition at 250 mM Aphanothece halophytica
tert-butylacetic acid 30% inhibition at 250 mM, with dimethylglycine Aphanothece halophytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
S-adenosyl-L-methionine pH 8.8, 37°C, recombinant wild-type enzyme, with dimethylglycine Aphanothece halophytica
0.25
-
S-adenosyl-L-methionine pH 8.8, 37°C, recombinant mutant P171Q, with dimethylglycine Aphanothece halophytica
0.5
-
dimethylglycine pH 8.8, 37°C, recombinant wild-type enzyme Aphanothece halophytica
0.6
-
S-adenosyl-L-methionine pH 8.8, 37°C, recombinant wild-type enzyme, with sarcosine Aphanothece halophytica
0.8
-
sarcosine pH 8.8, 37°C, recombinant wild-type enzyme Aphanothece halophytica
7.5
-
dimethylglycine pH 8.8, 37°C, recombinant mutant P171Q Aphanothece halophytica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29000
-
recombinant enzyme, gel filtration Aphanothece halophytica
31000
-
1 * 31000, recombinant enzyme, SDS-PAGE Aphanothece halophytica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Aphanothece halophytica the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress ?
-
?
S-adenosyl-L-methionine + N,N-dimethylglycine Aphanothece halophytica
-
S-adenosyl-L-homocysteine + betaine i.e. N,N,N-trimethylglycine ?
S-adenosyl-L-methionine + sarcosine Aphanothece halophytica
-
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?

Organism

Organism UniProt Comment Textmining
Aphanothece halophytica Q83WC3 inducible enzyme
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli by ammonium sulfate fractionation and ion exchange chromatography to homogeneity Aphanothece halophytica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress Aphanothece halophytica ?
-
?
additional information enzyme shows strict substrate specificity Aphanothece halophytica ?
-
?
S-adenosyl-L-methionine + glycine low activity Aphanothece halophytica S-adenosyl-L-homocysteine + sarcosine
-
?
S-adenosyl-L-methionine + N,N-dimethylglycine
-
Aphanothece halophytica S-adenosyl-L-homocysteine + betaine i.e. N,N,N-trimethylglycine ?
S-adenosyl-L-methionine + sarcosine
-
Aphanothece halophytica S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?
S-adenosyl-L-methionine + sarcosine i.e. methylglycine Aphanothece halophytica S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?

Subunits

Subunits Comment Organism
monomer 1 * 31000, recombinant enzyme, SDS-PAGE Aphanothece halophytica

Synonyms

Synonyms Comment Organism
SDMT
-
Aphanothece halophytica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Aphanothece halophytica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.8
-
assay at Aphanothece halophytica