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Literature summary extracted from

  • Kallnik, V.; Schultz, C.; Schweiger, P.; Deppenmeier, U.
    Properties of recombinant Strep-tagged and untagged hyperthermophilic D-arabitol dehydrogenase from Thermotoga maritima (2011), Appl. Microbiol. Biotechnol., 90, 1285-1293 .
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.1.250 synthesis the enzyme catalyzed oxidation of D-arabitol to D-ribulose which is a rare ketopentose sugar that has numerous industrially applications. D-Arabitol 2-dehydrogenase from Thermotoga maritima has the potential to be a useful biocatalyst for the production of D-ribulose starting from the inexpensive D-arabitol due to its regiospecificity and thermostability Thermotoga maritima

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.250 expressed in Escherichia coli Thermotoga maritima

General Stability

EC Number General Stability Organism
1.1.1.250 K+, 60 mM, twofold increase in catalytic rates. K+ likely contributes to the overall stability of the protein and is not required for catalytic activity Thermotoga maritima
1.1.1.250 Mg2+, 55 mM, twofold increase in catalytic rates. Mg2+ likely contributes to the overall stability of the protein and is not required for catalytic activity Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.250 0.027
-
 NADH 85°C, pH 7.5, Strep-tagged enzyme, cosubstrate D-ribulose Thermotoga maritima
1.1.1.250 0.031
-
 NADH 85°C, pH 7.5, tag-less enzyme, cosubstrate D-ribulose Thermotoga maritima
1.1.1.250 0.059
-
 NAD+ 85°C, pH 8.5, tag-less enzyme, cosubstrate D-arabitol Thermotoga maritima
1.1.1.250 0.069
-
 NAD+ 85°C, pH 8.5, Strep-tagged enzyme, cosubstrate D-arabitol Thermotoga maritima
1.1.1.250 2.6
-
 D-arabitol 85°C, pH 8.5, tag-less enzyme Thermotoga maritima
1.1.1.250 7.1
-
 D-arabitol 85°C, pH 8.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 7.2
-
 D-ribulose 85°C, pH 7.5, tag-less enzyme Thermotoga maritima
1.1.1.250 13.9
-
 D-ribulose 85°C, pH 7.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 20
-
 D-xylitol 85°C, pH 8.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 21.7
-
 D-xylitol 85°C, pH 8.5, tag-less enzyme Thermotoga maritima

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.250 K+ 60 mM, twofold increase in catalytic rates. K+ likely contributes to the overall stability of the protein and is not required for catalytic activity Thermotoga maritima
1.1.1.250 Mg2+ 55 mM, twofold increase in catalytic rates. Mg2+ likely contributes to the overall stability of the protein and is not required for catalytic activity Thermotoga maritima
1.1.1.250 additional information no effect on the activity is observed with CaCl2 or NaCl addition, up to 150 mM Thermotoga maritima

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.250 164000
-
 non-denaturing PAGE Thermotoga maritima
1.1.1.250 320000
-
 non-denaturing PAGE Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.250 Thermotoga maritima Q9WYD3
-
-
1.1.1.250 Thermotoga maritima DSM 3109 Q9WYD3
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.250 purified from Escherichia coli with and without a Strep-tag. The tag-less form of D-arabitol dehydrogenase has similar kinetic parameters compared to the tagged enzyme, demonstrating that the Strep-tag is not deleterious to protein function but decreases protein stability Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.250 D-arabitol + NAD+
-
 Thermotoga maritima D-ribulose + NADH + H+
-
 ?
1.1.1.250 D-arabitol + NAD+
-
 Thermotoga maritima DSM 3109 D-ribulose + NADH + H+
-
 ?
1.1.1.250 D-ribulose + NADH + H+
-
 Thermotoga maritima D-arabitol + NAD+
-
 ?
1.1.1.250 D-ribulose + NADH + H+
-
 Thermotoga maritima DSM 3109 D-arabitol + NAD+
-
 ?
1.1.1.250 D-xylitol + NAD+
-
 Thermotoga maritima D-xylulose + NADH + H+
-
 ?
1.1.1.250 D-xylitol + NAD+
-
 Thermotoga maritima DSM 3109 D-xylulose + NADH + H+
-
 ?
1.1.1.250 D-xylulose + NADH + H+
-
 Thermotoga maritima D-xylitol + NAD+
-
 ?
1.1.1.250 D-xylulose + NADH + H+
-
 Thermotoga maritima DSM 3109 D-xylitol + NAD+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.250 homododecamer 12 * 27600, SDS-PAGE Thermotoga maritima
1.1.1.250 homohexamer 6 * 27600, SDS-PAGE Thermotoga maritima

Synonyms

EC Number Synonyms Comment Organism
1.1.1.250 D-arabitol 2-dehydrogenase
-
 Thermotoga maritima
1.1.1.250 TM_0297
-
 Thermotoga maritima

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.250 85
-
 substrate: D-arabitol Thermotoga maritima

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.250 50
-
 inactive at temperatures lower than 50 °C Thermotoga maritima

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.250 85
-
 the tag-less protein is thermostable, retaining 90% of its activity after 90 min. The Strep-tagged enzyme loese 60% of its activity after 10 min Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.250 11
-
 D-arabitol 85°C, pH 8.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 11.2
-
 D-xylitol 85°C, pH 8.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 16.6
-
 D-xylitol 85°C, pH 8.5, tag-less enzyme Thermotoga maritima
1.1.1.250 18.7
-
 D-arabitol 85°C, pH 8.5, tag-less enzyme Thermotoga maritima
1.1.1.250 165.6
-
 D-ribulose 85°C, pH 7.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 193.2
-
 D-ribulose 85°C, pH 7.5, tag-less enzyme Thermotoga maritima

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.250 7.5
-
 reduction of D-ribulose Thermotoga maritima
1.1.1.250 8.5
-
 oxidation of D-arabitol Thermotoga maritima

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.250 4.5 9.5 oxidation of D-arabitol is not observed below pH 4.5 or above pH 9.5 Thermotoga maritima
1.1.1.250 5.5 8.5 pH 5.5: about 40% of maximal activity, pH 8.5: about 10% of maximal activity, reduction of D-ribulose Thermotoga maritima

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.250 NAD+ no activity with NADP+ Thermotoga maritima
1.1.1.250 NADH no activity with NADPH Thermotoga maritima

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.250 0.56
-
 D-xylitol 85°C, pH 8.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 0.74
-
 D-xylitol 85°C, pH 8.5, tag-less enzyme Thermotoga maritima
1.1.1.250 1.5
-
 D-arabitol 85°C, pH 8.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 7.2
-
 D-arabitol 85°C, pH 8.5, tag-less enzyme Thermotoga maritima
1.1.1.250 12
-
 D-ribulose 85°C, pH 7.5, Strep-tagged enzyme Thermotoga maritima
1.1.1.250 27
-
 D-ribulose 85°C, pH 7.5, tag-less enzyme Thermotoga maritima