Information on EC 1.1.1.365 - D-galacturonate reductase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.1.1.365
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RECOMMENDED NAME
GeneOntology No.
D-galacturonate reductase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-galactonate + NADP+ = D-galacturonate + NADPH + H+
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
D-galacturonate degradation III
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L-ascorbate biosynthesis V
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ascorbate metabolism
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degradation of sugar acids
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Pentose and glucuronate interconversions
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Ascorbate and aldarate metabolism
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
L-galactonate:NADP+ oxidoreductase
The enzyme from plants is involved in ascorbic acid (vitamin C) biosynthesis [1,2]. The enzyme from the fungus Trichoderma reesei (Hypocrea jecorina) is involved in a eukaryotic degradation pathway of D-galacturonate. It is also active with D-glucuronate and glyceraldehyde [3]. Neither enzyme shows any activity with NADH.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
a D-galacturonic acid-utilizing, psychrophilic yeast strain
HV538330
GenBank
Manually annotated by BRENDA team
a D-galacturonic acid-utilizing, psychrophilic yeast strain
HV538330
GenBank
Manually annotated by BRENDA team
cv. Guinong 5, a rose species with high ascorbic acid content
UniProt
Manually annotated by BRENDA team
anamorph Trichoderma reesei
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D-galactose + NADPH + H+
?
show the reaction diagram
D-galacturonate + NADH + H+
L-galactonate + NAD+
show the reaction diagram
D-galacturonate + NADPH + H+
L-galactonate + NADP+
show the reaction diagram
D-glucose + NADPH + H+
?
show the reaction diagram
D-glucuronate + NADH + H+
?
show the reaction diagram
D-glucuronate + NADPH + H+
?
show the reaction diagram
D-glucuronic acid + NADPH + H+
?
show the reaction diagram
D-xylose + NADPH + H+
?
show the reaction diagram
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29.9% activity compared to D-glucuronate
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?
DL-glyceraldehyde + NADPH + H+
?
show the reaction diagram
L-arabinose + NADPH + H+
?
show the reaction diagram
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14.2% activity compared to D-glucuronate
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ir
L-galactonate + NAD+
D-galacturonate + NADH + H+
show the reaction diagram
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r
L-galactonate + NADP+
D-galacturonate + NADPH + H+
show the reaction diagram
L-galactose + NADPH + H+
?
show the reaction diagram
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18.8% activity compared to D-glucuronate
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ir
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
D-galacturonate + NADPH + H+
L-galactonate + NADP+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cu2+
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135.3% activity at 5 mM
additional information
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not influenced by Ca2+, Co2+, Mn2+, Ni2+, Zn2+, and Fe2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
N-ethylmaleimide
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84% residual activity at 1 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
H2O2
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the enzyme is activated by 0.1 mM H2O2
additional information
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not influenced by EDTA, BCS, dithiothreiol, , and p-chloromercuribenzoate, L-GalL, and AsA
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.175 - 7.11
D-galacturonate
0.12 - 4.67
D-glucuronate
11
D-glucuronic acid
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in 100 mM sodium phosphate (pH 7.0), at 22C
8.48
D-xylose
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in 50 mM Tris-HCl (pH 7.2), at 25C
6
DL-glyceraldehyde
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in 100 mM sodium phosphate (pH 7.0), at 22C
4
L-galactonate
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in 100 mM sodium phosphate (pH 7.0), at 22C
0.326
NADH
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with D-galacturonate as cosubstrate, in 10 mM sodium phosphate, at pH 7.2 and 22C
0.001
NADP+
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in 100 mM sodium phosphate (pH 7.0), at 22C
0.03 - 0.0625
NADPH
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.19 - 6.88
D-galacturonate
0.13 - 3.11
D-glucuronate
0.06
D-xylose
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in 50 mM Tris-HCl (pH 7.2), at 25C
0.08 - 0.16
L-galactonate
4.98
NADH
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with D-galacturonate as cosubstrate, in 10 mM sodium phosphate, at pH 7.2 and 22C
7.11
NADPH
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with D-galacturonate as cosubstrate, in 10 mM sodium phosphate, at pH 7.2 and 22C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05095 - 39.29
D-galacturonate
0.04135
D-glucuronate
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in 50 mM Tris-HCl (pH 7.2), at 25C
0.00697
D-xylose
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in 50 mM Tris-HCl (pH 7.2), at 25C
15.72
NADH
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with D-galacturonate as cosubstrate, in 10 mM sodium phosphate, at pH 7.2 and 22C
197.2
NADPH
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with D-galacturonate as cosubstrate, in 10 mM sodium phosphate, at pH 7.2 and 22C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00422
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crude extract, at pH 7.2 and at 25C
0.12
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with NAD+ and L-galactonate as substrates, in 10 mM sodium phosphate, at pH 7.2 and 22C
0.17
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with NADH and D-glucuronate as substrates, in 10 mM sodium phosphate, at pH 7.2 and 22C
0.21
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with NADP+ and L-galactonate as substrates, in 10 mM sodium phosphate, at pH 7.2 and 22C
0.671
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after 159fold purification, at pH 7.2 and at 25C
3.9
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with NADPH and D-glucuronate as substrates, in 10 mM sodium phosphate, at pH 7.2 and 22C
6.9
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with NADH and D-galacturonate as substrates, in 10 mM sodium phosphate, at pH 7.2 and 22C
8.7
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with NADPH and D-galacturonate as substrates, in 10 mM sodium phosphate, at pH 7.2 and 22C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5
HV538330
in vivo activity
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
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about 75, 65 and 18.5% of maximum activity at pH 6.5, 8.0 and 9.0, respectively
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36900
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x * 36900, isoform GAR1, calculated from amino acid sequence
38000
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1 * 38000, SDS-PAGE
39000
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gel filtration
40000
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x * 40000, SDS-PAGE
44000
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x * 44000, His6-tagged enzyme, SDS-PAGE
49200
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x * 49200, isoform GAR2, calculated from amino acid sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CM-Sepharose column chromatography, ammonium sulfate precipitation, butyl Toyopearl column chromatography, and HiTrap Blue column chromatography
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Fractogel TSK DEAE-650 column chromatography
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glutathione agarose column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Arabidopsis thaliana and in Escherichia coli XL1-Blue MRF' cells
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expressed in Escherichia coli BL21 cells
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expressed in Escherichia coli DH5alpha cells
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expressed in Saccharomyces cerevisiae strain CEN.PK2-1B
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expressed in Solanum tuberosum cultivar Taedong Valley
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expressed in Solanum tuberosum via Agrobacterium tumefaciens-mediated transformation
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gene gaaA, expression analysis in wild-type strain and in a strain overexpressing the D-galacturonic acid-specific GatA transporter
gene GalUR, real-time quantitative PCR enzyme expression analysis
gene GalUR, recombinant overexpression in Solanum lycopersicum cv. Ailsa Craig leaves and fruits via Agrobacterium tumefaciens strain C58C1 transfection resulting in 2fold and 1.6fold increase in ascorbate level in tomato fruit and leaf, respectively, which correlates positively with FaGalUR transcriptional abundance and enzyme GalUR activity compared to wild-type plants, real-time PCR expression analysis
gene GalUR, recombinant stable overexpression in Solanum lycopersicum cv. H15 leaves and fruits using Agrobacterium tumefaciens strain EHA 105 transfection method, the pNW2300 vector, and the CaMV35S promoter. The transgenic plants show increased enzyme content and activity compared to wild-type. Semi-quantitative RT-PCR expression analysis, increased ascorbic acid content is correlated with high level of transcript expression, that is 2-3fold increased compared to wild-type
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gene GalUR, recombinant stable overexpression in Solanum lycopersicum cv. Zhongshu 4 leaves and fruits via Agrobacterium tumefaciens strain C58C1 transfection method using the pCB302-3 vector and CaMV 35S promoter, reverse transcriptase-PCR expression analysis. The transgenic plants show 2.2-2.5fold increased enzyme activity compared to wild-type
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gene GAR1, DNA and amino acid sequence determination and analysis, functional recombinant expression in Saccharomyces cerevisiae strain IFO 10455 from a genome integrated copy, integration of the pGK406GAR1 plasmid into the ura3 allele of the IFO 10455 genome, subcloning in Escherichia coli strain DH5alpha
HV538330
ORF FaGalUR is recombinantly expressed in Solanum lycopersicum from plasmid pPGGUSI, which contains a 4.8 Kb promoter fragment of the polygalacturonase (PG) gene and its terminator, or from a plasmid containing the 35S CaMV promoter, mobilized from Escherichia coli by triparental mating, via Agrobacterium tumefaciens strain LBA4404 in tomato plants, PCR expression analysis. FaGalUR expression driven by the CaMV 35S promoter causes the expression of the gene in most parts of the plants, while in the second, FaGalU, is expressed under the control of the tomato fruit ripening-specific PG promoter. All of the transgenic lines are morphologically indistinguishable over different generations from control lines both in vegetative traits, such as leaf size or plant height, and fruit traits such as color or size. The majority of transgenic plants display a slight increase in fruit yield, up to 1.4fold, which is a consequence of an increase in the number of fruits rather than an increase in fruit weight. No significant changes in soluble solids of transgenic plants, but a reduction in acidity
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
isoform GAR1 is induced about 10fold, while isoform GAR2 is strongly induced in cultures with D-galacturonic acid, pectate, and pectin as carbon source at 3 h after transfer, compared to continuous growth in glucose-containing culture
the enzyme is not regulated by L-galactono-1,4-lactone and ascorbate
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nutrition
accumulation of high levels of ascorbate using GalUR gene overepression in tomato fruits. Tomato fruits are considered a major dietary source of vitamin C in many countries, because it is consumed regularly and in large quantities. Tomato also serves as a fruit model for other crops species with fleshy berry. Accordingly, it is very important to monitor and to increase the vitamin C content in tomato fruit, for meeting the consumer demand and health requirements for high nutrition, e.g. by overexpressing D-galacturonate reductase