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Literature summary for 1.5.8.4 extracted from

  • Brizio, C.; Brandsch, R.; Bufano, D.; Pochini, L.; Indiveri, C.; Barile, M.
    Over-expression in Escherichia coli, functional characterization and refolding of rat dimethylglycine dehydrogenase (2004), Protein Expr. Purif., 37, 434-442.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Escherichia coli strain JM109 as N-terminally His-tagged enzyme being equally distributed in the soluble fraction and the insoluble inclusion body fraction Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
N,N-dimethylglycine pH 7.0, 30°C, recombinant holoenzyme, 2 Km-values Rattus norvegicus
9.4
-
N,N-dimethylglycine pH 7.0, 30°C, recombinant holoenzyme, 2 Km-values Rattus norvegicus

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
Rattus norvegicus 5739
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
90000
-
x * 90000, native enzyme, SDS-PAGE, x * 93000, recombinant enzyme, SDS-PAGE Rattus norvegicus
93000
-
x * 90000, native enzyme, SDS-PAGE, x * 93000, recombinant enzyme, SDS-PAGE Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5,10-methenyltetrahydrofolate + FADH2 Rattus norvegicus
-
5,10-methylene tetrahydrofolate + FAD
-
?
N,N-dimethylglycine + H2O + FAD Rattus norvegicus
-
sarcosine + formaldehyde + FADH2
-
?

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
male Wistar rats
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme from liver mitochondria, recombinant His-tagged enzyme from Escherichia coli 45fold by nickel affinity chromatography Rattus norvegicus

Renatured (Commentary)

Renatured (Comment) Organism
refolding by 80fold dilution after solubilization from inclusion bodies by urea treatment, 50-60% reactivation yield, overview Rattus norvegicus

Source Tissue

Source Tissue Comment Organism Textmining
liver
-
Rattus norvegicus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.24
-
purified recombinant enzyme from soluble fraction Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methenyltetrahydrofolate + FADH2
-
Rattus norvegicus 5,10-methylene tetrahydrofolate + FAD
-
?
N,N-dimethylglycine + H2O + FAD
-
Rattus norvegicus sarcosine + formaldehyde + FADH2
-
?
N,N-dimethylglycine + H2O + oxidized 1-methoxy-5-methylphenazinium methylsulfate
-
Rattus norvegicus sarcosine + formaldehyde + reduced 1-methoxy-5-methylphenazinium methylsulfate
-
?
N,N-dimethylglycine + H2O + oxidized 2,6-dichlorophenolindophenol
-
Rattus norvegicus sarcosine + formaldehyde + reduced 2,6-dichlorophenolindophenol
-
?

Subunits

Subunits Comment Organism
? x * 90000, native enzyme, SDS-PAGE, x * 93000, recombinant enzyme, SDS-PAGE Rattus norvegicus

Synonyms

Synonyms Comment Organism
Me2GlyDH
-
Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
recombinant and native enzyme Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
recombinant and native enzyme Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
FAD absolutely required, covalently bound to the apoenzyme via a histidinyl(N3)-(8alpha)FAD linkage, cofactor-free apoenzyme shows 70% reduced activity compared to the holoenzyme Rattus norvegicus
additional information 1-methoxy-5-methylphenazinium methylsulfate can act as electron acceptor Rattus norvegicus
additional information 2,6-dichlorophenolindophenol can act as electron acceptor Rattus norvegicus