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Literature summary for 2.4.1.82 extracted from
- Functional expression of a cDNA encoding pea (Pisum sativum L.) raffinose synthase, partial purification of the enzyme from maturing seeds, and steady-state kinetic analysis of raffinose synthesis (2002), Planta, 215, 839-846.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| functional expression as His-tagged protein in Spodoptera frugiperda Sf21 cells via baculovirus infection | Pisum sativum |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 1-deoxygalactonojirimycin | strong, competitive towards galactinol | Pisum sativum |  |
| myo-inositol | competitive against sucrose | Pisum sativum | |
| Ni2+ | destabilization, 30.2% activity at 1 mM | Pisum sativum | |
| sucrose | - |
Pisum sativum | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1 | - |
galactinol | hydrolysis | Pisum sativum | |
| 7.3 | - |
galactinol | partially purified native enzyme, raffinose synthesis | Pisum sativum | |
| 7.9 | - |
galactinol | recombinant enzyme in Sf21 cell extract, raffinose synthesis | Pisum sativum | |
| 22.6 | - |
sucrose | recombinant enzyme in Sf21 cell extract, raffinose synthesis | Pisum sativum | |
| 22.9 | - |
sucrose | partially purified native enzyme, raffinose synthesis | Pisum sativum | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 88700 | - |
amino acid sequence determination | Pisum sativum |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pisum sativum | Q8VWN6 | cv. Wunder von Kelvedon | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| no success in purifying the recombinant His-tagged enzyme from Sf21 insect cells | Pisum sativum |
| partial | Pisum sativum |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| alpha-D-galactosyl-(1->3)-1D-myo-inositol + sucrose = myo-inositol + raffinose | ping-pong reaction mechanism | Pisum sativum |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| seed | maturing | Pisum sativum | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.00005 | - |
recombinant in Sf21 insect cells | Pisum sativum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1-alpha-D-galactosyl-myo-inositol + D-ononitol | D-ononitol is 1-D-4-O-methyl-myo-inositol | Pisum sativum | myo-inositol + alpha-D-galactosyl-D-ononitol | - |
r | |
| 1-alpha-D-galactosyl-myo-inositol + D-pinitol | D-pinitol is 1-D-3-O-methyl-chiro-inositol | Pisum sativum | myo-inositol + galactopinitol A | i.e. O-alpha-D-galactopyranosyl-(1,2)-4-O-methyl-D-chiro-inositol | r | |
| 1-alpha-D-galactosyl-myo-inositol + H2O | hydrolysis, in the absence of acceptor substrate | Pisum sativum | myo-inositol + D-galactose | - |
? | |
| 1-alpha-D-galactosyl-myo-inositol + sucrose | - |
Pisum sativum | myo-inositol + raffinose | - |
r | |
| 4-nitrophenol-alpha-D-galactopyranoside + sucrose | - |
Pisum sativum | 4-nitrophenol + raffinose | - |
? | |
| alpha-D-galactosyl-D-ononitol + sucrose | - |
Pisum sativum | D-ononitol + raffinose | - |
? | |
| additional information | substrate specificity | Pisum sativum | ? | - |
? | |
| additional information | raffinose is no acceptor substrate | Pisum sativum | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| raffinose synthase | - |
Pisum sativum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Pisum sativum |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Pisum sativum |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.189 | - |
1-deoxygalactonojirimycin | - |
Pisum sativum | |
| 3.7 | - |
sucrose | hydrolysis reaction of galactinol, noncompetitive against galactinol | Pisum sativum | |
| 10.1 | - |
myo-inositol | competitive against sucrose | Pisum sativum | |
| 22.3 | - |
myo-inositol | noncompetitive against galactinol | Pisum sativum | |
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