Information on EC 2.1.1.156 - glycine/sarcosine N-methyltransferase

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The expected taxonomic range for this enzyme is: Bacteria, Archaea

EC NUMBER
COMMENTARY hide
2.1.1.156
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RECOMMENDED NAME
GeneOntology No.
glycine/sarcosine N-methyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + glycine = S-adenosyl-L-homocysteine + sarcosine
show the reaction diagram
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S-adenosyl-L-methionine + sarcosine = S-adenosyl-L-homocysteine + N,N-dimethylglycine
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glycine, serine and threonine metabolism
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glycine betaine biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:glycine(or sarcosine) N-methyltransferase [sarcosine(or N,N-dimethylglycine)-forming]
Cells of the oxygen-evolving halotolerant cyanobacterium Aphanocthece halophytica synthesize betaine from glycine by a three-step methylation process. This is the first enzyme and it leads to the formation of either sarcosine or N,N-dimethylglycine, which is further methylated to yield betaine (N,N,N-trimethylglycine) by the action of EC 2.1.1.157, sarcosine/dimethylglycine N-methyltransferase. Differs from EC 2.1.1.20, glycine N-methyltransferase, as it can further methylate the product of the first reaction. Acetate, dimethylglycine and S-adenosyl-L-homocysteine can inhibit the reaction [3].
CAS REGISTRY NUMBER
COMMENTARY hide
294210-82-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GSMT; the enzyme is encoded in the gene cluster of Mpgsmt-sdmt, gene Mpgsmt
UniProt
Manually annotated by BRENDA team
gene gsmA encodes a Gsm homologue protein
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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disruption of the gsmA gene, encoding Gsm homologue protein, generates a mutant that exhibits a longer lag period of growth and delayed spore germination under osmostress, it shows normal growth in CYE medium at 30C, phenotype, overview
metabolism
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glycine sarcosine methyltransferase and sarcosine dimethylglycine methyltransferase, EC 2.1.1.157, catalyze glycine betaine synthesis from glycine
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 S-adenosyl-L-methionine + glycine
2 S-adenosyl-L-homocysteine + N,N-dimethylglycine
show the reaction diagram
overall reaction
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?
S-adenosyl-L-methionine + glycine
S-adenosyl-L-homocysteine + N-methylglycine
show the reaction diagram
S-adenosyl-L-methionine + glycine
S-adenosyl-L-homocysteine + sarcosine
show the reaction diagram
S-adenosyl-L-methionine + N-methylglycine
S-adenosyl-L-homocysteine + N,N-dimethylglycine
show the reaction diagram
S-adenosyl-L-methionine + sarcosine
S-adenosyl-L-homocysteine + N,N-dimethylglycine
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + glycine
S-adenosyl-L-homocysteine + N-methylglycine
show the reaction diagram
S-adenosyl-L-methionine + glycine
S-adenosyl-L-homocysteine + sarcosine
show the reaction diagram
S-adenosyl-L-methionine + N-methylglycine
S-adenosyl-L-homocysteine + N,N-dimethylglycine
show the reaction diagram
S-adenosyl-L-methionine + sarcosine
S-adenosyl-L-homocysteine + N,N-dimethylglycine
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
dramatic activating effects of sodium and potassium ions on the in vitro methyltransferase activities of MpGSMT
Na+
dramatic activating effects of sodium and potassium ions on the in vitro methyltransferase activities of MpGSMT
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-chloromercuribenzoate
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recombinant enzyme: over 95% inhibition at 1.33 mM, completely reversible by 5.3 mM DTT
acetate
; 80% inhibition at 250 mM, with glycine
betaine
dimethylglycine
versus glycine, 65% inhibition at 250 mM
ethanolamine
; 45% inhibition at 250 mM, with glycine
KCl/NaCl
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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
monomethyl ethanolamine
; 35% inhibition at 250 mM, with glycine
N,N-dimethylglycine
phenylalanine
10% inhibition at 250 mM, with glycine
proline
; 30% inhibition at 250 mM, with glycine
S-adenosyl-L-homocysteine
sarcosine
competitive inhibition, versus glycine, 70% inhibition at 250 mM
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
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
betaine
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recombinant enzyme: 20% activation at 0.5 M with sarcosine, no activation with glycine
additional information
abiotic stess-inducible enzyme, e.g. high salt content
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1 - 2320
glycine
0.8 - 2.94
N-methylglycine
0.28 - 75
S-adenosyl-L-methionine
0.8 - 3020
sarcosine
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05 - 0.07
S-adenosyl-L-homocysteine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.023
activity with glycine and S-adenosyl-L-methionine in crude cell extracts
0.058
activity with glycine and S-adenosyl-L-methionine in crude cell extracts
0.075
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purified recombinant enzyme, substrate sarcosine
0.16
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purified recombinant enzyme, substrate glycine
0.19
activity with sarcosine and S-adenosyl-L-methionine, purified enzyme
0.23
activity with sarcosine and S-adenosyl-L-methionine in crude cell extracts
0.34
activity with sarcosine and S-adenosyl-L-methionine in crude cell extracts
0.52
activity with glycine and S-adenosyl-L-methionine, purified enzyme
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
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recombinant enzyme, with substrate glycine
7.9
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recombinant enzyme, with substrate sarcosine
8
with N-methylglycine as substrate
8.2
with glycine as substrate
9
glycine or N-methylglycine
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 11
glycine or N-methylglycine, very low activity below pH 8
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 7
sequence calculation
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
29040
2 * 29040, sequence calculation, 2 * 35000, SDS-PAGE, quaternary structure analysis of MpGSMT by analytical ultracentrifugation
29500
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x * 29500, about, sequence calculation
31000
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1 * 42000, recombinant enzyme, SDS-PAGE, 1 * 31000, about, amino acid sequence calculation
31210
calculated from the deduced amino acid sequence
32000
gel filtration; recombinant enzyme, gel filtration
33000
1 * 33000, recombinant enzyme, SDS-PAGE; 1 * 33000, SDS-PAGE, 1 * 32000, gel filtration
35500
gel filtration
40000
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gel filtration
42000
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1 * 42000, recombinant enzyme, SDS-PAGE, 1 * 31000, about, amino acid sequence calculation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 29500, about, sequence calculation
monomer
additional information
enzyme aggregated to tetramers is inactive
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme is instable during purification, activity is lost within 30 min
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; recombinant enzyme from Escherichia coli by ammonium sulfate fractionation and ion exchange chromatography to homogeneity
by hydrophobic interaction chromatography, ammonium sulfate fractionation, gel filtration, ion exchange chromatography and ultrafiltration
recombinant enzyme from Escherichia coli by ammonium sulfate fractionation, hydrophobic and anion exchange chromatography, and ultrafiltration to homogeneity
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recombinant MpGSMT from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant protein using His-tag
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination, functional coexpression in Escherichia coli with sarcosine dimethylglycine methyltransferase, EC 2.1.1.157, conferring betaine accumulation and increased salt tolerance to the Escherichia coli cells
DNA and amino acid sequence determination, the single gene encodes the glycine sarcosine methyltransferase as well as the sarcosine dimethylglycine methyltransferase, EC 2.1.1.157, the latter being probably a proteolytic processing product of the first, poor expression in Escherichia coli of the full length and the truncated enzyme
expressed as soluble His-tag fusion protein in Escherichia coli BL21(DE3)pLysS
expressed in Arabidopsis thaliana
expressed in Escherichia coli BL21(DE3); 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)
expressed in Oryza sativa cultivar Nipponbare
expressed in Zea mays
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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
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gene gsmA, DNA and amino acid sequence determination and analysis, sequence comparison
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gene Mpgsmt, DNA and amino acid sequence determination and analysis and sequence comparisons, phylogenetic analysis of GSMT, expression of complete gene cluster of Mpgsmt-sdmt in Escherichia coli strain BL21(DE3) and in strain MKH13 deficient in betaine transport, the recombinant enzyme is functional to synthesize and accumulate betaine and confers elevated survival ability in betaine transport deficient mutant strain MKH13 under high salt stress. The recombinantly expressed enzyme shows 10000fold lower catalytic efficiency compared with the enzyme isolated from the original cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
betaine downregulates GSMT expression enhanced with increasing temperature, transcription unit and transcription level of Mpgsmt-sdmt is reduced under temperature stresses up to 37C, but recovers at 45C to the level at 20C
exposure of embryos to benzo(a)pyrene causes increase in expression of mRNA
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the enzyme shows increased expression levels under drought conditions
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R169E
inactive; site-directed mutagenesis, completely inactive mutant
R169K
drastic increase of Km of the methyl-acceptor substrates, 2-fold increase in Km for S-adenosyl-L-methionine; site-directed mutagenesis, reduced activity and highly increased Km for ATP with both glycine and sarcosine compared to the wild-type enzyme
additional information
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construction of a chromosomal gsmA gene disruption knockout mutant, phenotype, overview
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
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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
biotechnology
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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
nutrition
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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
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