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Literature summary for 3.5.2.5 extracted from
- Tissue abundance and characterization of two purified proteins with allantoinase activity from French bean (Phaseolus vulgaris) (2007), Physiol. Plant., 131, 355-366.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| beta-mercaptoethanol | 14% increase of activity at 0.1 mM for IsoI and 11% increase of activity at 1 mM for IsoII | Phaseolus vulgaris |  |
| diethyl dicarbonate | 63% increase of activity at 0.1 mM for IsoI and 38% increase of activity at 1 mM for IsoII | Phaseolus vulgaris | |
| dithiothreitol | 20% increase of activity at 1 mM for IsoI and% increase of activity at 0.1 mM for IsoII | Phaseolus vulgaris | |
| phenyl phosphoramidate | 4% increase of activity at 1 mM for IsoI and 10% increase of activity at 1 mM for IsoII | Phaseolus vulgaris | |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-hydroxymercuribenzoate | inactivation, prevented by DTT | Phaseolus vulgaris | |
| Ca2+ | 62% residual activity at 1 mM for IsoI and 34% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
| Co2+ | 40% residual activity at 1 mM for IsoI and 30% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
| Cu2+ | 14% residual activity at 1 mM for IsoI and 8% residual activity at 1 mM for IsoII; high inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
| diethyl dicarbonate | 89% residual activity at 10 mM for IsoI and 90% residual activity at 10 mM for IsoII | Phaseolus vulgaris | |
| EDTA | 49% residual activity at 1 mM for IsoI and 40% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII, Fe2+ and Mn2+ restore the enzyme activity partially, while Ca2+ restores it completely | Phaseolus vulgaris | |
| Fe2+ | 75% residual activity at 1 mM for IsoI and 58% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
| iodoacetic acid | 16% residual activity at 5 mM for IsoI and 13% residual activity at 5 mM for IsoII | Phaseolus vulgaris | |
| Mg2+ | 44% residual activity at 1 mM for IsoI and 49% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
| Mn2+ | 81% residual activity at 1 mM for IsoI and 51% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
| additional information | the enzyme is inhibited by sulfhydryl-reactive agents | Phaseolus vulgaris | |
| N-ethylmaleimide | 54% residual activity at 5 mM for IsoI and 67% residual activity at 5 mM for IsoII | Phaseolus vulgaris | |
| Na+ | 51% residual activity at 1 mM for IsoI and 43% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
| p-hydroxymercuribenzoate | 10% residual activity at 0.5 mM for IsoI and 15% residual activity at 0.5 mM for IsoII | Phaseolus vulgaris | |
| Zn2+ | 29% residual activity at 1 mM for IsoI and 38% residual activity at 1 mM for IsoII; inhibition of isozymes IsoI and IsoII | Phaseolus vulgaris | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Phaseolus vulgaris | |
| 0.059 | - |
allantoin | pH 7.8, 40°C, isozyme IsoI, different methods | Phaseolus vulgaris | |
| 0.066 | - |
allantoin | pH 7.8, 40°C, isozyme IsoII, different methods | Phaseolus vulgaris | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mn2+ | - |
Phaseolus vulgaris | |
| additional information | metalloenzyme | Phaseolus vulgaris |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 42000 | - |
IsoII, native PAGE with acrylamide gradient | Phaseolus vulgaris |
| 42000 | - |
IsoII, SDS-PAGE, after purification two peaks of 45000 and 42000 Da length showing allantoinase activity are obtained | Phaseolus vulgaris |
| 45000 | - |
IsoI, SDS-PAGE, after purification two peaks of 45000 and 42000 Da length showing allantoinase activity are obtained | Phaseolus vulgaris |
| 53000 | - |
IsoI, native PAGE with acrylamide gradient | Phaseolus vulgaris |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| allantoin + H2O | Phaseolus vulgaris | a key reaction step in the biosynthesis and degradation of ureides, overview | allantoate | - |
? | |
Organic Solvent Stability
| Organic Solvent | Comment | Organism |
|---|---|---|
| deoxycholate | 20% increase of activity in the presence of 0.1% (w/v) deoxycholate | Phaseolus vulgaris |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Phaseolus vulgaris | - |
- |
- |
| Phaseolus vulgaris | - |
cv. Great Northern, two isozymes IsoI and IsoII | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| DEAE-Sephacel column chromatography, MonoQ HR column chromatography, and Sephacryl S-300 gel filtration | Phaseolus vulgaris |
| native isozymes 1 and 2 from developing fruits 150fold and 286fold, respectively, to homogeneity involving heat treatment at 55°C for 20 min, anion exchange chromatography, gel filtration, and another step of anion exchange chromatography | Phaseolus vulgaris |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| fruit | highest activity | Phaseolus vulgaris | - |
| fruit | developing, high enzyme expression level | Phaseolus vulgaris | - |
| leaf | - |
Phaseolus vulgaris | - |
| leaf | IsoI is the predominant form in all the tissues analysed except leaves, where most of the allantoinase activity corresponds to IsoII enzyme | Phaseolus vulgaris | - |
| additional information | tissue distribution, expression especially during fertilization, ubiquitous expression of isozymes in bean tissues, one isozyme is the main form in leaves and the other isozyme is the main form in roots, overview | Phaseolus vulgaris | - |
| root | - |
Phaseolus vulgaris | - |
| root | the amount of IsoII is very low in roots | Phaseolus vulgaris | - |
| seedling | - |
Phaseolus vulgaris | - |
| shoot | - |
Phaseolus vulgaris | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
activity of isozymes in different bean tissues and during development, overview | Phaseolus vulgaris |
| 2 | - |
crude extract | Phaseolus vulgaris |
| 295 | - |
purified isozyme IsoII | Phaseolus vulgaris |
| 400 | - |
after 200fold purification | Phaseolus vulgaris |
| 563 | - |
purified isozyme IsoI | Phaseolus vulgaris |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-allantoin + H2O | - |
Phaseolus vulgaris | allantoate | - |
? | |
| allantoin + H2O | - |
Phaseolus vulgaris | allantoate | - |
? | |
| allantoin + H2O | a key reaction step in the biosynthesis and degradation of ureides, overview | Phaseolus vulgaris | allantoate | - |
? | |
| additional information | no activity with hydantoin and 5-bromouracil | Phaseolus vulgaris | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | - |
Phaseolus vulgaris |
| monomer | 1 * 42000, isozyme IsoII, SDS-PAGE, 1 * 45000, isozyme IsoI, SDS-PAGE | Phaseolus vulgaris |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| allantoin amidohydrolase | - |
Phaseolus vulgaris |
| IsoI | 45000 Da enzyme | Phaseolus vulgaris |
| IsoII | 42000 Da enzyme | Phaseolus vulgaris |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | - |
- |
Phaseolus vulgaris |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | 70 | the activity of both protein variants IsoI and IsoII is very resistant to heat treatment because the enzyme is fully active after treatment for 30 min at 60°C and shows more than 50% activity after 30 min at 70°C | Phaseolus vulgaris |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 201.7 | - |
allantoin | pH 7.8, 40°C, isozyme IsoII, different methods | Phaseolus vulgaris | |
| 420 | - |
allantoin | pH 7.8, 40°C, isozyme IsoI, different methods | Phaseolus vulgaris | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.7 | - |
different methods | Phaseolus vulgaris |
| 7.8 | - |
different methods | Phaseolus vulgaris |
| 7.8 | - |
IsoI | Phaseolus vulgaris |
| 8.1 | - |
IsoII | Phaseolus vulgaris |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 8 | - |
Phaseolus vulgaris |
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