Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Song, S.H.; Ahluwalia, N.; Leduc, Y.; Delbaere, L.T.; Vieille, C.
    Thermotoga maritima TM0298 is a highly thermostable mannitol dehydrogenase (2008), Appl. Microbiol. Biotechnol., 81, 485-495.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.67 expressed in Escherichia coli BL21(DE3) cells Thermotoga maritima
1.1.1.67 MtDH is expressed in Escherichia coli BL21(DE3) Limosilactobacillus reuteri
1.1.1.67 the TM0298 gene subcloned into the NdeI and XhoI sites of pET24a(+) to yield plasmid pTmMtDH, from which MtDH is expressed with a C-terminal His6-tag in Escherichia coli BL21(DE3) Thermotoga maritima

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.67 EDTA activity of MtDH pretreated with 10 mM EDTA for 20 min at 37°C decreases 96%. Adding 20 mM ZnCl2 to the EDTA-treated enzyme restores activity up to 80% of the control (enzyme not treated with EDTA). Adding 20 mM CoCl2 and 20 mM MnCl2 to the EDTA-treated enzyme restores activity up to 132% and 94% of the control, respectively. In contrast, adding 20 mM MgCl2 or CaCl2 does not increase the activity of the EDTA-treated enzyme.; activity of MtDH pretreated with 10 mM for 20 min at 37°C decreases 96%. Adding 20 mM ZnCl2 to the EDTA-treated enzyme restores activity up to 80% of the control. Adding 20 mM CoCl2 and 20 mM MnCl2 to the EDTA-treated enzyme restores activity up to 132% and 94% of the control, respectively Thermotoga maritima
1.1.1.67 mannitol
-
 Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.67 0.037
-
 NADH at 60°C and pH 6.1 Thermotoga maritima
1.1.1.67 0.037
-
 NADH at 60°C, pH 6.1 Thermotoga maritima
1.1.1.67 0.048
-
 NADH at 80°C and pH 6.1 Thermotoga maritima
1.1.1.67 0.048
-
 NADH at 80°C, pH 6.1 Thermotoga maritima
1.1.1.67 0.14
-
 NAD+ at 80°C and pH 8.3 Thermotoga maritima
1.1.1.67 0.14
-
 NAD+ at 80°C, pH 8.3 Thermotoga maritima
1.1.1.67 0.17
-
 NADPH at 80°C and pH 6.1 Thermotoga maritima
1.1.1.67 0.17
-
 NADPH at 80°C, pH 6.1 Thermotoga maritima
1.1.1.67 5.51
-
 D-mannitol at 80°C and pH 8.3 Thermotoga maritima
1.1.1.67 5.51
-
 D-mannitol at 80°C, pH 8.3 Thermotoga maritima
1.1.1.67 7.5
-
 NADP+ at 80°C and pH 8.3 Thermotoga maritima
1.1.1.67 7.5
-
 NADP+ at 80°C, pH 8.3 Thermotoga maritima
1.1.1.67 13.23
-
 D-mannitol at 60°C and pH 6.1 Thermotoga maritima
1.1.1.67 13.23
-
 D-mannitol at 60°C, pH 6.1 Thermotoga maritima
1.1.1.67 50
-
 D-fructose at 60°C and pH 6.1 Thermotoga maritima
1.1.1.67 50
-
 D-fructose at 60°C, pH 6.1 Thermotoga maritima
1.1.1.67 50.97
-
 D-fructose at 80°C and pH 6.1 Thermotoga maritima
1.1.1.67 50.97
-
 D-fructose at 80°C, pH 6.1 Thermotoga maritima

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.67 Co2+
-
 Thermotoga maritima
1.1.1.67 Mn2+
-
 Thermotoga maritima
1.1.1.67 additional information 20 mM MgCl2 or CaCl2 do not increase the activity of the EDTA-treated enzyme Thermotoga maritima
1.1.1.67 Zn2+ contains 0.69 mol of Zn2+ per subunit of enzyme Thermotoga maritima
1.1.1.67 Zn2+ MtDH contains a single catalytic zinc per subunit Thermotoga maritima

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.67 34000
-
 4 * 34000, SDS-PAGE Thermotoga maritima
1.1.1.67 120000
-
 ultracentrifugation Thermotoga maritima
1.1.1.67 120000 143000 gel filtration or analytical ultracentrifugation Thermotoga maritima
1.1.1.67 120000 134000 gel filtration Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.67 Limosilactobacillus reuteri Q6ECH5
-
-
1.1.1.67 Thermotoga maritima Q9WYD4
-
-
1.1.1.67 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 Q9WYD4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.67 by heat treatment at 85°C and on Ni-NTA column Thermotoga maritima
1.1.1.67 Ni-NTA agarose column chromatography Thermotoga maritima

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.67 54
-
 purified enzyme at 80°C with D-fructose as the substrate and NADH as the cofactor Thermotoga maritima
1.1.1.67 54
-
 purified enzyme, at 80°C with fructose as the substrate and NADH as the cofactor Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.67 D-fructose + NADPH + H+ also active on fructose with NADPH Thermotoga maritima D-mannitol + NADP+
-
 ?
1.1.1.67 D-fructose + NADPH + H+ also active on fructose with NADPH Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 D-mannitol + NADP+
-
 ?
1.1.1.67 D-mannitol + NAD(P)+
-
 Thermotoga maritima D-fructose + NAD(P)H + H+
-
 r
1.1.1.67 D-mannitol + NAD(P)+
-
 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 D-fructose + NAD(P)H + H+
-
 r
1.1.1.67 D-mannitol + NAD+ 100% activity Thermotoga maritima D-fructose + NADH + H+
-
 r
1.1.1.67 D-mannitol + NAD+ 100% activity Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 D-fructose + NADH + H+
-
 r
1.1.1.67 D-tagatose + NAD(P)+ 29% relative activity compared to D-fructose Thermotoga maritima ? + NAD(P)H
-
 ?
1.1.1.67 D-tagatose + NAD+ 29% relative activity on D-tagatose compared to 100% activity on D-fructose Thermotoga maritima ?
-
 ?
1.1.1.67 D-xylulose + NAD(P)+ 18% relative activity compared to D-fructose Thermotoga maritima ? + NAD(P)H
-
 ?
1.1.1.67 D-xylulose + NAD+ 18% relative activity on D-xylulose compared to 100% activity on D-fructose Thermotoga maritima ?
-
 ?
1.1.1.67 L-sorbose + NAD(P)+ 5% relative activity compared to D-fructose Thermotoga maritima ? + NAD(P)H
-
 ?
1.1.1.67 L-sorbose + NADH + H+ 5% relative activity on L-sorbose compared to 100% activity on D-fructose Thermotoga maritima L-sorbitol + NAD+
-
 ?
1.1.1.67 additional information mannitol can be produced directly from glucose in a two-step enzymatic process, using a Thermotoga neapolitana xylose isomerase mutant and TmMtDH at 60°C. No activity with glucose, xylose, threonine, arabinose, acetaldehyde, 2-butanone, sorbitol, xylitol, ethanol, or 2-butanol Thermotoga maritima ?
-
 ?
1.1.1.67 additional information TM0298 shows no detectable activity on glucose, xylose, threonine, arabinose, acetaldehyde, 2-butanone, sorbitol, xylitol, ethanol, or 2-butanol Thermotoga maritima ?
-
 ?
1.1.1.67 additional information mannitol can be produced directly from glucose in a two-step enzymatic process, using a Thermotoga neapolitana xylose isomerase mutant and TmMtDH at 60°C. No activity with glucose, xylose, threonine, arabinose, acetaldehyde, 2-butanone, sorbitol, xylitol, ethanol, or 2-butanol Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 ?
-
 ?
1.1.1.67 additional information TM0298 shows no detectable activity on glucose, xylose, threonine, arabinose, acetaldehyde, 2-butanone, sorbitol, xylitol, ethanol, or 2-butanol Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.67 dimer gel filtration at room temperature, MtDH in solution Limosilactobacillus reuteri
1.1.1.67 octamer gel filtration at room temperature, MtDH in solution Limosilactobacillus reuteri
1.1.1.67 tetramer 4 * 34000, SDS-PAGE Thermotoga maritima
1.1.1.67 tetramer gel filtration at room temperature, MtDH in solution Limosilactobacillus reuteri
1.1.1.67 tetramer gel filtration or analytical ultracentrifugation, MtDH in solution Thermotoga maritima

Synonyms

EC Number Synonyms Comment Organism
1.1.1.67 MtDH
-
 Thermotoga maritima
1.1.1.67 MtDH
-
 Limosilactobacillus reuteri
1.1.1.67 TM0298
-
 Thermotoga maritima

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.67 90 100 most active around 95°C Thermotoga maritima
1.1.1.67 95
-
-
 Thermotoga maritima

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.67 90 120 MtDH retains 63% of its activity at 120°C but shows no detectable activity at 25°C Thermotoga maritima
1.1.1.67 95 120 retains 63% of its activity at 120°C but shows no detectable activity at room temperature Thermotoga maritima

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.67 80
-
 half-life: 57 min Thermotoga maritima
1.1.1.67 80
-
 half-lives of 57 min Thermotoga maritima
1.1.1.67 95
-
 half-life: 6 min Thermotoga maritima
1.1.1.67 95
-
 half-lives of 6 min Thermotoga maritima

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.67 additional information
-
 pH 5.5-6 is the optimum for D-fructose reduction and pH 8.3-8.6 is the optimum for mannitol reduction Thermotoga maritima
1.1.1.67 5.5 6 optimally reduces L-fructose at pH values between pH 5.5 and 6.0 Thermotoga maritima
1.1.1.67 8.3 8.6 optimally oxidizes L-mannitol between pH 8.3 and 8.6 Thermotoga maritima

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.67 8.5
-
 TM0298 is sensitive to proteolytic degradation in purification protocols at pH 8.5 Thermotoga maritima

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.67 NAD(P)+ its catalytic efficiency is 33times higher with NAD+ than with NADP+ Thermotoga maritima
1.1.1.67 NAD(P)H MtDH has a higher Vmax with NADPH than with NADH, whereas its catalytic efficiency is 2.2times higher with NADH than with NADPH. Cofactor specificity is due to the high density of negatively charged residues (Glu193, Asp195, and Glu196) downstream of the NAD(P) interaction site, the glycine motif Thermotoga maritima
1.1.1.67 NAD+ although MtDH has a higher Vmax with NADPH than with NADH, its catalytic efficiency is 33 times higher with NAD+ than with NADP+ Thermotoga maritima
1.1.1.67 NADH although MtDH has a higher Vmax with NADPH than with NADH, its catalytic efficiency is 2.2times higher with NADH than with NADPH Thermotoga maritima

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.67 99
-
 mannitol
-
 Thermotoga maritima