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Literature summary for 2.1.1.156 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
expressed in Escherichia coli BL21(DE3) Aphanothece halophytica
orf1, 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 sarcosine dimethylglycine methyltransferase, EC 2.1.1.157, increases the intracellular betaine level and the growth rate of Escherichia coli strain BL21(DE3) Aphanothece halophytica

Protein Variants

Protein Variants Comment Organism
R169E inactive Aphanothece halophytica
R169E site-directed mutagenesis, completely inactive mutant Aphanothece halophytica
R169K site-directed mutagenesis, reduced activity and highly increased Km for ATP with both glycine and sarcosine compared to the wild-type enzyme Aphanothece halophytica
R169K drastic increase of Km of the methyl-acceptor substrates, 2-fold increase in Km for S-adenosyl-L-methionine Aphanothece halophytica

Inhibitors

Inhibitors Comment Organism Structure
acetate 80% inhibition at 250 mM, with glycine Aphanothece halophytica
dimethylglycine versus glycine, 65% inhibition at 250 mM Aphanothece halophytica
ethanolamine 45% inhibition at 250 mM, with glycine Aphanothece halophytica
KCl
-
Aphanothece halophytica
monomethyl ethanolamine 35% inhibition at 250 mM, with glycine Aphanothece halophytica
additional information inhibition rates with sarcosine, overview, no feedback inhibition by betaine up to 2 M, no inhibition with glycine by serine, methionine, asparagine, glutamate, cysteine, and alanine; no inhibition with betaine Aphanothece halophytica
N,N-dimethylglycine
-
Aphanothece halophytica
NaCl
-
Aphanothece halophytica
phenylalanine 10% inhibition at 250 mM, with glycine Aphanothece halophytica
proline 30% inhibition at 250 mM, with glycine Aphanothece halophytica
S-adenosyl-L-homocysteine product inhibition Aphanothece halophytica
sarcosine competitive inhibition, versus glycine, 70% inhibition at 250 mM Aphanothece halophytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
S-adenosyl-L-methionine pH 8.8, 37°C, recombinant wild-type enzyme, with sarcosine Aphanothece halophytica
0.6
-
S-adenosyl-L-methionine with N-methylglycine as methyl group acceptor, pH 8.8, 37°C Aphanothece halophytica
0.7
-
S-adenosyl-L-methionine pH 8.8, 37°C, recombinant wild-type enzyme, with glycine Aphanothece halophytica
0.7
-
S-adenosyl-L-methionine with glycine as methyl group acceptor, pH 8.8, 37°C Aphanothece halophytica
0.8
-
N-methylglycine pH 8.8, 37°C Aphanothece halophytica
0.8
-
sarcosine pH 8.8, 37°C, recombinant wild-type enzyme Aphanothece halophytica
1
-
glycine pH 8.8, 37°C, recombinant mutant R169K Aphanothece halophytica
1.1
-
sarcosine pH 8.8, 37°C, recombinant mutant R169K Aphanothece halophytica
1.5
-
glycine pH 8.8, 37°C Aphanothece halophytica
1.5
-
glycine pH 8.8, 37°C, recombinant wild-type enzyme Aphanothece halophytica
50
-
S-adenosyl-L-methionine pH 8.8, 37°C, recombinant mutant R169K, with sarcosine Aphanothece halophytica
75
-
S-adenosyl-L-methionine pH 8.8, 37°C, recombinant mutant R169K, with glycine Aphanothece halophytica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
31210
-
calculated from the deduced amino acid sequence Aphanothece halophytica
32000
-
gel filtration Aphanothece halophytica
32000
-
recombinant enzyme, gel filtration Aphanothece halophytica
33000
-
1 * 33000, recombinant enzyme, SDS-PAGE Aphanothece halophytica
33000
-
1 * 33000, SDS-PAGE, 1 * 32000, gel filtration 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 + glycine Aphanothece halophytica
-
S-adenosyl-L-homocysteine + sarcosine
-
?
S-adenosyl-L-methionine + glycine Aphanothece halophytica synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine S-adenosyl-L-homocysteine + N-methylglycine
-
?
S-adenosyl-L-methionine + N-methylglycine Aphanothece halophytica synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?
S-adenosyl-L-methionine + sarcosine Aphanothece halophytica
-
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?

Organism

Organism UniProt Comment Textmining
Aphanothece halophytica
-
-
-
Aphanothece halophytica Q83WC4 inducible enzyme
-

Purification (Commentary)

Purification (Comment) Organism
-
Aphanothece halophytica
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
-
Aphanothece halophytica S-adenosyl-L-homocysteine + sarcosine
-
?
S-adenosyl-L-methionine + glycine
-
Aphanothece halophytica S-adenosyl-L-homocysteine + sarcosine i.e. 2-methylglycine ?
S-adenosyl-L-methionine + glycine
-
Aphanothece halophytica S-adenosyl-L-homocysteine + N-methylglycine
-
?
S-adenosyl-L-methionine + glycine synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine Aphanothece halophytica S-adenosyl-L-homocysteine + N-methylglycine
-
?
S-adenosyl-L-methionine + N-methylglycine
-
Aphanothece halophytica S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?
S-adenosyl-L-methionine + N-methylglycine synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine Aphanothece halophytica S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?
S-adenosyl-L-methionine + sarcosine
-
Aphanothece halophytica S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
?

Subunits

Subunits Comment Organism
monomer 1 * 33000, recombinant enzyme, SDS-PAGE Aphanothece halophytica
monomer 1 * 33000, SDS-PAGE, 1 * 32000, gel filtration Aphanothece halophytica
More enzyme aggregated to tetramers is inactive Aphanothece halophytica

Synonyms

Synonyms Comment Organism
GSMT
-
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
9
-
glycine or N-methylglycine Aphanothece halophytica

pH Range

pH Minimum pH Maximum Comment Organism
8 11 glycine or N-methylglycine, very low activity below pH 8 Aphanothece halophytica