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Literature summary for 1.1.1.138 extracted from

  • Sasaki, Y.; Laivenieks, M.; Zeikus, J.G.
    Lactobacillus reuteri ATCC 53608 mdh gene cloning and recombinant mannitol dehydrogenase characterization (2005), Appl. Microbiol. Biotechnol., 68, 36-41.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene mdh, DNA and amino acid sequence determination and compiational amnipulation, expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Limosilactobacillus reuteri

Inhibitors

Inhibitors Comment Organism Structure
EDTA 93% inhibition at 10 mM within 20 min, reversible by Zn2+ which restores 66% of control activity at 20 mM Limosilactobacillus reuteri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.04
-
NADPH pH 5.4, 37°C, recombinant enzyme Limosilactobacillus reuteri
0.24
-
NADH pH 5.4, 37°C, recombinant enzyme Limosilactobacillus reuteri
0.35
-
NADP+ pH 6.2, 37°C, recombinant enzyme Limosilactobacillus reuteri
9
-
NAD+ pH 6.2, 37°C, recombinant enzyme Limosilactobacillus reuteri
34
-
D-fructose pH 5.4, 37°C, recombinant enzyme Limosilactobacillus reuteri
54
-
D-mannitol pH 6.2, 37°C, recombinant enzyme Limosilactobacillus reuteri

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ content determination at 20°C, 0.87 mol Zn2+ per subunit Limosilactobacillus reuteri

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37279
-
2 * 40000, recombinant enzyme, SDS-PAGE, 2 * 37279, recombinant His6-tagged enzyme, mass spectrometry Limosilactobacillus reuteri
40000
-
2 * 40000, recombinant enzyme, SDS-PAGE, 2 * 37279, recombinant His6-tagged enzyme, mass spectrometry Limosilactobacillus reuteri
75000
-
recombinant enzyme, gel filtration Limosilactobacillus reuteri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-mannitol + NAD(P)+ Limosilactobacillus reuteri
-
D-fructose + NAD(P)H + H+
-
r

Organism

Organism UniProt Comment Textmining
Limosilactobacillus reuteri
-
strain ATCC 53608, gene mdh
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultratfiltration Limosilactobacillus reuteri

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.61
-
crude recombinant Escherichia coli cell extract Limosilactobacillus reuteri
124.6
-
purified recombinant enzyme Limosilactobacillus reuteri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-mannitol + NAD(P)+
-
Limosilactobacillus reuteri D-fructose + NAD(P)H + H+
-
r
D-mannitol + NAD+
-
Limosilactobacillus reuteri D-fructose + NADH + H+
-
?

Subunits

Subunits Comment Organism
dimer 2 * 40000, recombinant enzyme, SDS-PAGE, 2 * 37279, recombinant His6-tagged enzyme, mass spectrometry Limosilactobacillus reuteri

Synonyms

Synonyms Comment Organism
LRMDH
-
Limosilactobacillus reuteri
mannitol dehydrogenase
-
Limosilactobacillus reuteri
MDH
-
Limosilactobacillus reuteri

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
recombinant enzyme Limosilactobacillus reuteri

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
purified recombinant enzyme, 5 min, complete inactivation Limosilactobacillus reuteri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4
-
reverse reaction, recombinant enzyme Limosilactobacillus reuteri
6.2
-
forward reaction, recombinant enzyme Limosilactobacillus reuteri

pH Range

pH Minimum pH Maximum Comment Organism
5 7 reverse reaction, 81% of maximal activity at pH 5.0, 22% at pH 7.0, 72% at pH 5.6, and 60% at pH 7.4 Limosilactobacillus reuteri

Cofactor

Cofactor Comment Organism Structure
NAD(P)+ NADP+ is the preferred cofactor in the forward reaction Limosilactobacillus reuteri
NAD(P)H NADPH is the preferred cofactor in the reverse reaction Limosilactobacillus reuteri