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Literature summary extracted from

  • Gonzalez-Porque, P.; Strominger, J.L.
    Enzymatic synthesis of cytidine diphosphate 3,6-dideoxyhexoses. VI. Purification to homogeneity and some properties of cytidine diphosphate-D-glucose oxidoreductase, enzyme E1 and enzyme E3 (1972), J. Biol. Chem., 247, 6748-6756.
    View publication on PubMed

General Stability

EC Number General Stability Organism
4.2.1.45 unstable in dilute salt solutions Yersinia pseudotuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.17.1.1 5,5'-dithiobis(2-nitrobenzoate)
-
 Yersinia pseudotuberculosis
1.17.1.1 N-ethylmaleimide enzyme E1: dithiothreitol in excess protects against inhibition Yersinia pseudotuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.17.1.1 35000 45000 enzyme E3, thin-layer chromatography on Sephadex G-100 Yersinia pseudotuberculosis
1.17.1.1 40000
-
 enzyme E3: 1 * 40000, SDS-PAGE, enzyme E1: 1 * 61000, SDS-PAGE, 2 proteins E1 and E3 are involved but no partial reaction has been observed in the presence of either alone Yersinia pseudotuberculosis
1.17.1.1 50000 70000 enzyme E1, thin-layer chromatography on Sephadex G-100 Yersinia pseudotuberculosis
1.17.1.1 61000
-
 enzyme E3: 1 * 40000, SDS-PAGE, enzyme E1: 1 * 61000, SDS-PAGE, 2 proteins E1 and E3 are involved but no partial reaction has been observed in the presence of either alone Yersinia pseudotuberculosis
4.2.1.45 43000
-
 2 * 43000, SDS-PAGE Yersinia pseudotuberculosis
4.2.1.45 70000 100000 gel filtration Yersinia pseudotuberculosis

Organism

EC Number Organism UniProt Comment Textmining
1.17.1.1 Yersinia pseudotuberculosis
-
 25 VO
-
1.17.1.1 Yersinia pseudotuberculosis 25VO
-
 25 VO
-
4.2.1.45 Yersinia pseudotuberculosis
-
 Pasteurella pseudotuberculosis
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.17.1.1 the first three steps are common to the purification of enzyme E1, enzyme E3 and cofactor, their separation can be accomplished after step 4, step 1: preparation of the crude extract, step 2: streptomycin sulfate precipitation, step 3: ammonium sulfate precipitation and dialysis, step 4: DEAE-cellulose chromatography, step 5: purification of enzyme E1, gel filtration on Sephadex G-100, step 6: second DEAE-cellulose chromatography, step 7: preparative polyacrylamide gel electrophoresis, step 5': purification of enzyme E3, gel filtration on Sephadex G-100, step 6': second DEAE-cellulose chromatography, step 7': third DEAE-cellulose chromatography Yersinia pseudotuberculosis
4.2.1.45 partial Yersinia pseudotuberculosis

Reaction

EC Number Reaction Comment Organism Reaction ID
1.17.1.1 CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O = CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H + H+ mechanism Yersinia pseudotuberculosis
a
1.17.1.1 CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O = CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H + H+ E1 binds the substrate pyridoxamine 5'-phosphate essential for binding, E3 possesses NADH oxidase activity, may be the reductase Yersinia pseudotuberculosis
a
1.17.1.1 CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O = CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H + H+ role of the enzymes E1 and E3, as well as role of the cofactor. Multicomponent system consisting of substrate, NADH or NADPH, enzyme E1: recognition unit of the system by binding to the substrate through the amino group of the coenzyme, enzyme E3: acts as the dehydrogenase of the system, and cofactor: pyridoxamine 5'-phosphate Yersinia pseudotuberculosis
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.17.1.1 additional information
-
-
 Yersinia pseudotuberculosis

Storage Stability

EC Number Storage Stability Organism
1.17.1.1 lyophilized powder: enzyme E3 not stable Yersinia pseudotuberculosis
1.17.1.1 lyophilized powder: for months, enzyme E1 Yersinia pseudotuberculosis
4.2.1.45 -20°C, several months, no loss of activity Yersinia pseudotuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.45 CDP-D-glucose
-
 Yersinia pseudotuberculosis CDP-4-dehydro-6-deoxy-D-glucose + H2O
-
 ?
4.2.1.45 CDP-D-glucose
-
 Yersinia pseudotuberculosis 25VO CDP-4-dehydro-6-deoxy-D-glucose + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.17.1.1 monomer enzyme E3: 1 * 40000, SDS-PAGE, enzyme E1: 1 * 61000, SDS-PAGE, 2 proteins E1 and E3 are involved but no partial reaction has been observed in the presence of either alone Yersinia pseudotuberculosis
4.2.1.45 dimer 2 * 43000, SDS-PAGE Yersinia pseudotuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.17.1.1 25
-
 assay at Yersinia pseudotuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.17.1.1 7.5
-
 assay at Yersinia pseudotuberculosis

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
4.2.1.45 5.5
-
 not stable below Yersinia pseudotuberculosis

Cofactor

EC Number Cofactor Comment Organism Structure
1.17.1.1 additional information 4.3 SH per mol of enzyme E1 in the presence of SDS, calculated using a molecular weight of 61000 Da Yersinia pseudotuberculosis
1.17.1.1 additional information 0.37 SH groups per molecule of enzyme E3, in the absence of SDS and 1 SH group per molecule of enzyme E3 in the presence of SDS, the single SH group in enzyme E3 is essential for activity Yersinia pseudotuberculosis
1.17.1.1 pyridoxamine 5'-phosphate bound to enzyme E1 through an ionic interaction with a positive charge on the surface of the enzyme, the cofactor is needed for the binding of the substrate to the enzyme Yersinia pseudotuberculosis