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Literature summary extracted from

  • Ren, Y.; Perepelov, A.V.; Wang, H.; Zhang, H.; Knirel, Y.A.; Wang, L.; Chen, W.
    Biochemical characterization of GDP-L-fucose de novo synthesis pathway in fungus Mortierella alpina (2010), Biochem. Biophys. Res. Commun., 391, 1663-1669.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.271 DTT 2.1fold activation Mortierella alpina
4.2.1.47 dithiothreitol enhances GMD activity by 2.3fold Mortierella alpina
4.2.1.47 DTT enhances GMD activity by 2.3fold Mortierella alpina

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.271 expression of the His-tagged enzyme in Escherichia coli strain BL21 (DE3) Mortierella alpina
4.2.1.47 into pET28a+ to construct pLW1534, expressed in Escherichia coli BL21 as His-tagged fusion protein Mortierella alpina
4.2.1.47 recombinant expression of His-tagged GMD in Escherichia coli Mortierella alpina

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.271 Co2+
-
 Mortierella alpina
1.1.1.271 Cu2+ complete inhibition at 5 mM Mortierella alpina
1.1.1.271 Fe2+
-
 Mortierella alpina
1.1.1.271 Mn2+
-
 Mortierella alpina
4.2.1.47 Co2+ 5 mM, strong inhibition; strong inhibition at 5 mM Mortierella alpina
4.2.1.47 Cu2+ 5 mM, strong inhibition; strong inhibition at 5 mM Mortierella alpina
4.2.1.47 Fe2+ 5 mM, strong inhibition; strong inhibition at 5 mM Mortierella alpina
4.2.1.47 additional information is not affected by Ca2+ and Mn2+ Mortierella alpina

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.271 additional information
-
 additional information Michaelis-Menten kinetiic modeling Mortierella alpina
1.1.1.271 1.025
-
 NADPH recombinant His-tagged enzyme, pH 8.0, 37°C Mortierella alpina
1.1.1.271 1.047
-
 GDP-4-dehydro-6-deoxy-alpha-D-mannose recombinant His-tagged enzyme, pH 8.0, 37°C Mortierella alpina
1.1.1.271 1.157
-
 NADH recombinant His-tagged enzyme, pH 8.0, 37°C Mortierella alpina
4.2.1.47 0.77
-
 GDP-mannose pH 8.0, 37°C Mortierella alpina
4.2.1.47 0.77
-
 GDP-mannose recombinant enzyme, pH 8.0, 37°C Mortierella alpina

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.271 Ca2+ slight activation at 5 mM Mortierella alpina
1.1.1.271 Mg2+ slight activation at 5 mM Mortierella alpina
1.1.1.271 additional information no requirement of divalent cations Mortierella alpina
4.2.1.47 Mg2+ 5 mM, activity is significantly enhanced Mortierella alpina
4.2.1.47 Mg2+ activates at 5 mM Mortierella alpina
4.2.1.47 additional information divalent cations are not required for GMD activity Mortierella alpina
4.2.1.47 additional information GMD activity is not affected by Ca2+ at 5 mM Mortierella alpina

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.271 40000
-
 x * 40000, recombinant His-tagged enzyme, SDS-PAGE Mortierella alpina
4.2.1.47 41200
-
 3 * 45000, recombinant His-tagged GMD, SDS-PAGE, 3 * 41200, about, sequence calculation Mortierella alpina
4.2.1.47 45000
-
 3 * 45000, SDS-PAGE Mortierella alpina
4.2.1.47 45000
-
 3 * 45000, recombinant His-tagged GMD, SDS-PAGE, 3 * 41200, about, sequence calculation Mortierella alpina
4.2.1.47 140000
-
 gel filtration Mortierella alpina
4.2.1.47 140000
-
 recombinant His-tagged GMD, native PAGE Mortierella alpina

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.271 GDP-4-dehydro-6-deoxy-alpha-D-mannose + NADPH + H+ Mortierella alpina
-
 GDP-beta-L-fucose + NADP+
-
 ?
4.2.1.47 GDP-mannose Mortierella alpina
-
 GDP-4-keto-6-deoxy mannose + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.271 Mortierella alpina D3JU54
-
-
4.2.1.47 Mortierella alpina D3JU53
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.271 recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography Mortierella alpina
4.2.1.47 recombinant His-tagged GMD from Escherichia coli by nickel affinity chromatography Mortierella alpina
4.2.1.47 to near homogeneity by nickel ion affinity chromatography Mortierella alpina

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.271 GDP-4-dehydro-6-deoxy-alpha-D-mannose + NADH + H+ the enzyme synthesizes GDP-L-fucose from its substrate GDP-4-keto-6-deoxy-D-mannose. The reaction involves epimerization at both C-3 and C-5 followed by an NADPH-dependent reduction of the carbonyl at C-4 Mortierella alpina GDP-beta-L-fucose + NAD+ product identification by capillary electrophoresis, electro-spray ionization-mass spectrometry, and nuclear magnetic resonance spectroscopy ?
1.1.1.271 GDP-4-dehydro-6-deoxy-alpha-D-mannose + NADPH + H+
-
 Mortierella alpina GDP-beta-L-fucose + NADP+
-
 ?
1.1.1.271 GDP-4-dehydro-6-deoxy-alpha-D-mannose + NADPH + H+ the enzyme synthesizes GDP-L-fucose from its substrate GDP-4-keto-6-deoxy-D-mannose. The reaction involves epimerization at both C-3 and C-5 followed by an NADPH-dependent reduction of the carbonyl at C-4 Mortierella alpina GDP-beta-L-fucose + NADP+ product identification by capillary electrophoresis, electro-spray ionization-mass spectrometry, and nuclear magnetic resonance spectroscopy ?
4.2.1.47 GDP-mannose
-
 Mortierella alpina GDP-4-dehydro-6-deoxy-D-mannose + H2O
-
 ?
4.2.1.47 GDP-mannose
-
 Mortierella alpina GDP-4-keto-6-deoxy mannose + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.271 ? x * 40000, recombinant His-tagged enzyme, SDS-PAGE Mortierella alpina
4.2.1.47 trimer 3 * 45000, SDS-PAGE Mortierella alpina
4.2.1.47 trimer 3 * 45000, recombinant His-tagged GMD, SDS-PAGE, 3 * 41200, about, sequence calculation Mortierella alpina

Synonyms

EC Number Synonyms Comment Organism
1.1.1.271 GDP-keto-6-deoxymannose 3,5-epimerase/4-reductase
-
 Mortierella alpina
1.1.1.271 GMER
-
 Mortierella alpina
4.2.1.47 GDP-D-mannose 4,6-dehydratase
-
 Mortierella alpina
4.2.1.47 GMD
-
 Mortierella alpina

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.271 37
-
 recombinant His-tagged enzyme Mortierella alpina
4.2.1.47 37
-
-
 Mortierella alpina
4.2.1.47 37
-
 recombinant enzyme Mortierella alpina

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.271 15 55 activity range Mortierella alpina
4.2.1.47 25 55 25°C: about 60% of maximal activity, 55°C: about 70% of maximal activity Mortierella alpina

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.47 5.96
-
 GDP-mannose pH 8.0, 37°C Mortierella alpina
4.2.1.47 5.96
-
 GDP-mannose recombinant enzyme, pH 8.0, 37°C Mortierella alpina

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.271 7
-
 recombinant His-tagged enzyme Mortierella alpina
4.2.1.47 9
-
-
 Mortierella alpina
4.2.1.47 9
-
 recombinant enzyme Mortierella alpina

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.271 5 11 activity range Mortierella alpina
4.2.1.47 5 11 pH 5: 45% of maximal activity, pH 11: 75% of maximal activity Mortierella alpina

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.271 additional information both NADH and NADPH may be used as the cofactor by the enzyme, which shows similar affinities to NADH and NADPH, but the Vmax value is higher for NADPH Mortierella alpina
1.1.1.271 NADH
-
 Mortierella alpina
1.1.1.271 NADPH
-
 Mortierella alpina
4.2.1.47 NAD+
-
 Mortierella alpina
4.2.1.47 NADP+
-
 Mortierella alpina

General Information

EC Number General Information Comment Organism
4.2.1.47 metabolism GMD catalyzes the first step in the de novo synthesis pathways of GDP-L-fucose, overview Mortierella alpina