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Literature summary extracted from

  • Nyyssola, A.; Reinikainen, T.; Leisola, M.
    Characterization of glycine sarcosine N-methyltransferase and sarcosine dimethylglycine N-methyltransferase (2001), Appl. Environ. Microbiol., 67, 2044-2050.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.1.1.156 betaine recombinant enzyme: 20% activation at 0.5 M with sarcosine, no activation with glycine Halorhodospira halochloris

Application

EC Number Application Comment Organism
2.1.1.156 agriculture enzyme can be used in betaine production for improvement of stress tolerance of commercially important microbes in agriculture and industry, and for nutritial improvement of transgenic crop plants, that do not produce betaine naturally Halorhodospira halochloris
2.1.1.156 biotechnology enzyme can be used in betaine production for improvement of stress tolerance of commercially important microbes in agriculture and industry, and for nutritial improvement of transgenic crop plants, that do not produce betaine naturally Halorhodospira halochloris
2.1.1.156 nutrition enzyme can be used in betaine production for improvement of stress tolerance of commercially important microbes in agriculture and industry, and for nutritial improvement of transgenic crop plants, that do not produce betaine naturally Halorhodospira halochloris
2.1.1.157 agriculture enzyme can be used in betaine production for improvement of stress tolerance of commercially important microbes in agriculture and industry, and for nutritial improvement of transgenic crop plants, that do not produce betaine naturally Halorhodospira halochloris
2.1.1.157 biotechnology enzyme can be used in betaine production for improvement of stress tolerance of commercially important microbes in agriculture and industry, and for nutritial improvement of transgenic crop plants, that do not produce betaine naturally Halorhodospira halochloris
2.1.1.157 nutrition enzyme can be used in betaine production for improvement of stress tolerance of commercially important microbes in agriculture and industry, and for nutritial improvement of transgenic crop plants, that do not produce betaine naturally Halorhodospira halochloris

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.156 functional overexpression in Escherichia coli strain XL-1 Blue conferring betaine accumulation and increased salt tolerance when coexpressed with the sarcosine dimethylglycine methyltransferase, EC 2.1.1.157 Halorhodospira halochloris
2.1.1.157 functional overexpression in Escherichia coli strain XL-1 Blue conferring betaine accumulation and increased salt tolerance when coexpressed with the glycine sarcosine methyltransferase, EC 2.1.1.156 Halorhodospira halochloris

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.156 4-chloromercuribenzoate recombinant enzyme: over 95% inhibition at 1.33 mM, completely reversible by 5.3 mM DTT Halorhodospira halochloris
2.1.1.156 betaine recombinant enzyme: poor inhibition, 60% inhibition at 2 M with glycine, and 70% inhibition at 2 M with sarcosine, synergistic with salts KCl and NaCl, overview Halorhodospira halochloris
2.1.1.156 KCl/NaCl recombinant enzyme: synergistic with betaine, 80% inhibition at 0.33 M KCl and 0.66M NaCl with glycine, and 66% inhibition at 0.33 M KCl and 0.66 M NaCl with sarcosine, overview Halorhodospira halochloris
2.1.1.156 N,N-dimethylglycine recombinant enzyme: poor competitive product inhibition, 50% inhibition at 38 mM with glycine, and at 49 mM with sarcosine Halorhodospira halochloris
2.1.1.156 S-adenosyl-L-homocysteine recombinant enzyme: competitive product inhibition, 50% inhibition at 0.5 mM with glycine, and at 0.4 mM with sarcosine Halorhodospira halochloris
2.1.1.157 4-chloromercuribenzoate recombinant enzyme: 23% inhibition at 1.33 mM, completely reversible by 5.3 mM DTT Halorhodospira halochloris
2.1.1.157 betaine recombinant enzyme: poor inhibition, 71% inhibition at 2 M with sarcosine, and 69% inhibition at 2 M with dimethylglycine, partially reversible by salts KCl and NaCl, overview Halorhodospira halochloris
2.1.1.157 KCl/NaCl recombinant enzyme: 32% inhibition at 0.33 M KCl and 0.66 M NaCl with sarcosine, and 25% inhibition at 0.33 M KCl and 0.66 M NaCl with dimethylglycine, decrease inhibition by betaine, overview Halorhodospira halochloris
2.1.1.157 additional information no inhibition by N,N-dimethylglycine Halorhodospira halochloris
2.1.1.157 S-adenosyl-L-homocysteine recombinant enzyme: competitive product inhibition, 50% inhibition at 0.5 mM with glycine, and at 2.3 mM with sarcosine Halorhodospira halochloris

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.156 0.28
-
 S-adenosyl-L-methionine recombinant enzyme, pH 7.4, 37°C, with sarcosine Halorhodospira halochloris
2.1.1.156 0.42
-
 S-adenosyl-L-methionine recombinant enzyme, pH 7.4, 37°C, with glycine Halorhodospira halochloris
2.1.1.156 2.3
-
 sarcosine recombinant enzyme, pH 7.4, 37°C, with S-adenosyl-L-methionine Halorhodospira halochloris
2.1.1.156 18
-
 glycine recombinant enzyme, pH 7.4, 37°C, with S-adenosyl-L-methionine Halorhodospira halochloris
2.1.1.157 0.16
-
 S-adenosyl-L-methionine recombinant enzyme, pH 7.4, 37°C, with glycine Halorhodospira halochloris
2.1.1.157 0.21
-
 S-adenosyl-L-methionine recombinant enzyme, pH 7.4, 37°C, with sarcosine Halorhodospira halochloris
2.1.1.157 4.9
-
 dimethylglycine recombinant enzyme, pH 7.4, 37°C, with S-adenosyl-L-methionine Halorhodospira halochloris
2.1.1.157 6.1
-
 sarcosine recombinant enzyme, pH 7.4, 37°C, with S-adenosyl-L-methionine Halorhodospira halochloris

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.1.1.156 additional information no requirement for Ca2+, Mg2+, EDTA, Mn2+, Co2+, or Zn2+ Halorhodospira halochloris
2.1.1.157 additional information no requirement for Ca2+, Mg2+, EDTA, Mn2+, Co2+, or Zn2+ Halorhodospira halochloris

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.1.156 31000
-
 1 * 42000, recombinant enzyme, SDS-PAGE, 1 * 31000, about, amino acid sequence calculation Halorhodospira halochloris
2.1.1.156 40000
-
 gel filtration Halorhodospira halochloris
2.1.1.156 42000
-
 1 * 42000, recombinant enzyme, SDS-PAGE, 1 * 31000, about, amino acid sequence calculation Halorhodospira halochloris
2.1.1.157 25000
-
 gel filtration Halorhodospira halochloris
2.1.1.157 32000
-
 1 * 36000, recombinant enzyme, SDS-PAGE, 1 * 32000, about, amino acid sequence calculation Halorhodospira halochloris
2.1.1.157 36000
-
 1 * 36000, recombinant enzyme, SDS-PAGE, 1 * 32000, about, amino acid sequence calculation Halorhodospira halochloris

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.156 additional information Halorhodospira halochloris the enzyme catalyzes the first 2 steps of glycine betaine synthesis from glycine in a 3-step process, betaine is required for balancing osmotic pressure under high salt living conditions ?
-
 ?
2.1.1.156 S-adenosyl-L-methionine + glycine Halorhodospira halochloris
-
 S-adenosyl-L-homocysteine + sarcosine i.e. 2-methylglycine ?
2.1.1.156 S-adenosyl-L-methionine + sarcosine Halorhodospira halochloris
-
 S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
 ?
2.1.1.157 additional information Halorhodospira halochloris the enzyme catalyzes the last 2 steps of betaine synthesis from glycine in a 3-step process, betaine is required for balancing osmotic pressure under high salt living conditions ?
-
 ?
2.1.1.157 S-adenosyl-L-methionine + N,N-dimethylglycine Halorhodospira halochloris
-
 S-adenosyl-L-homocysteine + betaine
-
 ?
2.1.1.157 S-adenosyl-L-methionine + sarcosine Halorhodospira halochloris
-
 S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.156 Halorhodospira halochloris
-
 i.e. Halorhodospira halochloris
-
2.1.1.157 Halorhodospira halochloris
-
 i.e. Halorhodospira halochloris
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.156 recombinant enzyme from Escherichia coli by ammonium sulfate fractionation, hydrophobic and anion exchange chromatography, and ultrafiltration to homogeneity Halorhodospira halochloris
2.1.1.157 recombinant enzyme from Escherichia coli by ammonium sulfate fractionation, hydrophobic and anion exchange chromatography, and ultrafiltration to homogeneity Halorhodospira halochloris

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.1.156 additional information
-
-
 Halorhodospira halochloris
2.1.1.156 0.075
-
 purified recombinant enzyme, substrate sarcosine Halorhodospira halochloris
2.1.1.156 0.16
-
 purified recombinant enzyme, substrate glycine Halorhodospira halochloris
2.1.1.157 additional information
-
-
 Halorhodospira halochloris
2.1.1.157 0.68
-
 purified recombinant enzyme, substrate sarcosine Halorhodospira halochloris
2.1.1.157 3.4
-
 purified recombinant enzyme, substrate dimethylglycine Halorhodospira halochloris

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.156 additional information the enzyme catalyzes the first 2 steps of glycine betaine synthesis from glycine in a 3-step process, betaine is required for balancing osmotic pressure under high salt living conditions Halorhodospira halochloris ?
-
 ?
2.1.1.156 additional information the enzyme shows strict specificity for glycine and sarcosine as substrates, no activity with ethanolamine, monomethylethanolamine, nor several L-amino acid, overview Halorhodospira halochloris ?
-
 ?
2.1.1.156 S-adenosyl-L-methionine + glycine
-
 Halorhodospira halochloris S-adenosyl-L-homocysteine + sarcosine i.e. 2-methylglycine ?
2.1.1.156 S-adenosyl-L-methionine + glycine the recombinant enzyme from Escherichia coli is 3.3fold less active than the wild-type enzyme Halorhodospira halochloris S-adenosyl-L-homocysteine + sarcosine i.e. 2-methylglycine ?
2.1.1.156 S-adenosyl-L-methionine + sarcosine
-
 Halorhodospira halochloris S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
 ?
2.1.1.156 S-adenosyl-L-methionine + sarcosine the recombinant enzyme from Escherichia coli is 2.5fold less active than the wild-type enzyme Halorhodospira halochloris S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
 ?
2.1.1.157 additional information the enzyme catalyzes the last 2 steps of betaine synthesis from glycine in a 3-step process, betaine is required for balancing osmotic pressure under high salt living conditions Halorhodospira halochloris ?
-
 ?
2.1.1.157 additional information the enzyme shows strict specificity for glycine and sarcosine as substrates, no activity with ethanolamine, monomethylethanolamine, nor several L-amino acid, overview Halorhodospira halochloris ?
-
 ?
2.1.1.157 S-adenosyl-L-methionine + N,N-dimethylglycine
-
 Halorhodospira halochloris S-adenosyl-L-homocysteine + betaine
-
 ?
2.1.1.157 S-adenosyl-L-methionine + sarcosine
-
 Halorhodospira halochloris S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
 ?
2.1.1.157 S-adenosyl-L-methionine + sarcosine i.e. methylglycine Halorhodospira halochloris S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.1.156 monomer 1 * 42000, recombinant enzyme, SDS-PAGE, 1 * 31000, about, amino acid sequence calculation Halorhodospira halochloris
2.1.1.157 monomer 1 * 36000, recombinant enzyme, SDS-PAGE, 1 * 32000, about, amino acid sequence calculation Halorhodospira halochloris

Synonyms

EC Number Synonyms Comment Organism
2.1.1.156 glycine sarcosine N-methyltransferase
-
 Halorhodospira halochloris
2.1.1.156 GSMT
-
 Halorhodospira halochloris
2.1.1.157 sarcosine dimethylglycine N-methyltransferase
-
 Halorhodospira halochloris
2.1.1.157 SDMT
-
 Halorhodospira halochloris

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.1.156 37
-
 assay at Halorhodospira halochloris
2.1.1.157 37
-
 assay at Halorhodospira halochloris

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.156 7.4
-
 recombinant enzyme, with substrate glycine Halorhodospira halochloris
2.1.1.156 7.9
-
 recombinant enzyme, with substrate sarcosine Halorhodospira halochloris
2.1.1.157 additional information
-
 pH-optimum depends on the buffer system Halorhodospira halochloris
2.1.1.157 7.6
-
 recombinant enzyme, with substrate dimethylglycine, triethanolamine buffer Halorhodospira halochloris
2.1.1.157 8 9 recombinant enzyme, with substrate sarcosine, pH-optimum depends on the buffer system Halorhodospira halochloris