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Literature summary extracted from

  • Ishikawa, T.; Masumoto, I.; Iwasa, N.; Nishikawa, H.; Sawa, Y.; Shibata, H.; Nakamura, A.; Yabuta, Y.; Shigeoka, S.
    Functional characterization of D-galacturonic acid reductase, a key enzyme of the ascorbate biosynthesis pathway, from Euglena gracilis (2006), Biosci. Biotechnol. Biochem., 70, 2720-2726.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.365 H2O2 the enzyme is activated by 0.1 mM H2O2 Euglena gracilis
1.1.1.365 additional information not influenced by EDTA, BCS, dithiothreiol, , and p-chloromercuribenzoate, L-GalL, and AsA Euglena gracilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.365 N-ethylmaleimide 84% residual activity at 1 mM Euglena gracilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.365 0.0625
-
 NADPH in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis
1.1.1.365 3.79
-
 D-galacturonate in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis
1.1.1.365 4.67
-
 D-glucuronate in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis
1.1.1.365 8.48
-
 D-xylose in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.365 Cu2+ 135.3% activity at 5 mM Euglena gracilis
1.1.1.365 additional information not influenced by Ca2+, Co2+, Mn2+, Ni2+, Zn2+, and Fe2+ Euglena gracilis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.365 38000
-
 1 * 38000, SDS-PAGE Euglena gracilis
1.1.1.365 39000
-
 gel filtration Euglena gracilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.365 D-galacturonate + NADPH + H+ Euglena gracilis
-
 L-galactonate + NADP+
-
 ir
1.1.1.365 D-galacturonate + NADPH + H+ Euglena gracilis Z
-
 L-galactonate + NADP+
-
 ir

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.365 Euglena gracilis
-
-
-
1.1.1.365 Euglena gracilis Z
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.365 CM-Sepharose column chromatography, ammonium sulfate precipitation, butyl Toyopearl column chromatography, and HiTrap Blue column chromatography Euglena gracilis

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.365 0.00422
-
 crude extract, at pH 7.2 and at 25°C Euglena gracilis
1.1.1.365 0.671
-
 after 159fold purification, at pH 7.2 and at 25°C Euglena gracilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.365 D-galactose + NADPH + H+ 14.9% activity compared to D-glucuronate Euglena gracilis ?
-
 ir
1.1.1.365 D-galactose + NADPH + H+ 14.9% activity compared to D-glucuronate Euglena gracilis Z ?
-
 ir
1.1.1.365 D-galacturonate + NADH + H+ 17.7% activity compared to NADPH Euglena gracilis L-galactonate + NAD+
-
 ir
1.1.1.365 D-galacturonate + NADPH + H+
-
 Euglena gracilis L-galactonate + NADP+
-
 ir
1.1.1.365 D-galacturonate + NADPH + H+ 100% activity Euglena gracilis L-galactonate + NADP+
-
 ir
1.1.1.365 D-galacturonate + NADPH + H+
-
 Euglena gracilis Z L-galactonate + NADP+
-
 ir
1.1.1.365 D-glucose + NADPH + H+ 14.6% activity compared to D-glucuronate Euglena gracilis ?
-
 ir
1.1.1.365 D-glucose + NADPH + H+ 14.6% activity compared to D-glucuronate Euglena gracilis Z ?
-
 ir
1.1.1.365 D-glucuronate + NADPH + H+ 82.4% activity compared to D-glucuronate Euglena gracilis ?
-
 ?
1.1.1.365 D-glucuronate + NADPH + H+ 82.4% activity compared to D-glucuronate Euglena gracilis Z ?
-
 ?
1.1.1.365 D-xylose + NADPH + H+ 29.9% activity compared to D-glucuronate Euglena gracilis ?
-
 ?
1.1.1.365 DL-glyceraldehyde + NADPH + H+ 14.2% activity compared to D-glucuronate Euglena gracilis ?
-
 ir
1.1.1.365 L-arabinose + NADPH + H+ 14.2% activity compared to D-glucuronate Euglena gracilis ?
-
 ir
1.1.1.365 L-galactose + NADPH + H+ 18.8% activity compared to D-glucuronate Euglena gracilis ?
-
 ir
1.1.1.365 additional information the enzyme does not catalyze the reverse reaction with L-galactonate and NADP+ and does not utilize 4-nitrobenzaldehyde or menadione Euglena gracilis ?
-
 ?
1.1.1.365 additional information the enzyme does not catalyze the reverse reaction with L-galactonate and NADP+ and does not utilize 4-nitrobenzaldehyde or menadione Euglena gracilis Z ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.365 monomer 1 * 38000, SDS-PAGE Euglena gracilis

Synonyms

EC Number Synonyms Comment Organism
1.1.1.365 D-galacturonic acid reductase
-
 Euglena gracilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.365 0.06
-
 D-xylose in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis
1.1.1.365 0.19
-
 D-glucuronate in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis
1.1.1.365 0.19
-
 D-galacturonate in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.365 7.2
-
-
 Euglena gracilis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.365 6.5 9 about 75, 65 and 18.5% of maximum activity at pH 6.5, 8.0 and 9.0, respectively Euglena gracilis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.365 NADPH the enzyme prefers NADPH over NADH (ratio of 10 to 1.8) Euglena gracilis

Expression

EC Number Organism Comment Expression
1.1.1.365 Euglena gracilis the enzyme is not regulated by L-galactono-1,4-lactone and ascorbate additional information

General Information

EC Number General Information Comment Organism
1.1.1.365 metabolism D-galacturonic acid reductase is a key enzyme of the ascorbate biosynthesis pathway Euglena gracilis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.365 0.00697
-
 D-xylose in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis
1.1.1.365 0.04135
-
 D-glucuronate in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis
1.1.1.365 0.05095
-
 D-galacturonate in 50 mM Tris-HCl (pH 7.2), at 25°C Euglena gracilis