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Literature summary for 2.4.1.82 extracted from

  • Gangl, R.; Behmueller, R.; Tenhaken, R.
    Molecular cloning of AtRS4, a seed specific multifunctional RFO synthase/galactosylhydrolase in Arabidopsis thaliana (2015), Front. Plant Sci., 6, 789.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.4.1.82

Cloned(Commentary)

Cloned (Comment) Organism
gene RFS4, cloning from seeds and functional recombinant expression of His-tagged enzyme in Escherichia coli Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
additional information isolation of a T-DNA insertional mutant in the AtRS4 gene, only semi-quantitative PCR from wild-type siliques shows a specific transcriptional AtRS4PCR product. Metabolite measurements in seeds of DELTAAtRS4 mutant plants reveal a total loss of stachyose in DELTAAtRS4 mutant seeds Arabidopsis thaliana

General Stability

General Stability Organism
Mg2+, bovine serum albumin, Triton, Tween 20, and DTT stabilize the enzyme, the latter is used in the enzyme assay Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Arabidopsis thaliana
1.17
-
 alpha-D-galactosyl-(1->3)-1D-myo-inositol pH 7.0, 25°C, recombinant His-tagged enzyme Arabidopsis thaliana

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
98010
-
 x * 98010, about, sequence calculation, x * 100000, recombinant His-tagged enzyme, SDS-PAGE Arabidopsis thaliana
100000
-
 x * 98010, about, sequence calculation, x * 100000, recombinant His-tagged enzyme, SDS-PAGE Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose Arabidopsis thaliana
-
 myo-inositol + raffinose
-
 ?
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose Arabidopsis thaliana Col-0
-
 myo-inositol + raffinose
-
 ?
additional information Arabidopsis thaliana the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase ?
-
 ?
additional information Arabidopsis thaliana Col-0 the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase ?
-
 ?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q9SYJ4 gene RFS4, isozyme RS4
-
Arabidopsis thaliana Col-0 Q9SYJ4 gene RFS4, isozyme RS4
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography Arabidopsis thaliana

Source Tissue

Source Tissue Comment Organism Textmining
seed isozyme RS4 is seed-specific Arabidopsis thaliana
-
silique
-
 Arabidopsis thaliana
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose
-
 Arabidopsis thaliana myo-inositol + raffinose
-
 ?
alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose
-
 Arabidopsis thaliana Col-0 myo-inositol + raffinose
-
 ?
additional information the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase Arabidopsis thaliana ?
-
 ?
additional information the multifunctional enzyme is a raffinose and high affinity stachyose synthase, EC 2.4.1.82 and EC 2.4.1.67, as well as stachyose and galactinol specific galactosylhydrolase Arabidopsis thaliana Col-0 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 98010, about, sequence calculation, x * 100000, recombinant His-tagged enzyme, SDS-PAGE Arabidopsis thaliana

Synonyms

Synonyms Comment Organism
More cf. EC 2.4.1.67 Arabidopsis thaliana
RafS
-
 Arabidopsis thaliana
RFO synthase/galactosylhydrolase
-
 Arabidopsis thaliana
RS4
-
 Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Arabidopsis thaliana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Arabidopsis thaliana

General Information

General Information Comment Organism
malfunction metabolite phenotype of DELTAAtRS4 mutant seeds, overview Arabidopsis thaliana
metabolism biosynthesis of RFOs proceeds by stepwise transfer of galactosyl units. The second step involves raffinose synthase, AtRS5, EC 2.4.1.82, which transfers the galactosyl unit from galactinol to the C6 position of the glucose unit in sucrose forming an alpha-1,6-galactosidic linkage to yield the trisaccharide raffinose. In a third step, stachyose synthase, AtRS4, EC 2.4.1.67, transfers the galactosyl moiety from galactinol to the C6 position of the galactose unit in raffinose to yield the tetrasaccharide stachyose Arabidopsis thaliana
physiological function AtRS4 is the only stachyose synthase in the genome of Arabidopsis thaliana. It represents a key regulation mechanism in the raffinose family oligosaccharide physiology of Arabidosis thaliana due to its multifunctional enzyme activity. AtRS4 is possibly the second seed-specific raffinose synthase beside AtRS5, which is responsible for raffinose accumulation under abiotic stress Arabidopsis thaliana