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Literature summary extracted from

  • Raso, M.J.; Pineda, M.; Piedras, P.
    Tissue abundance and characterization of two purified proteins with allantoinase activity from French bean (Phaseolus vulgaris) (2007), Physiol. Plant., 131, 355-366.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.2.5 beta-mercaptoethanol 14% increase of activity at 0.1 mM for IsoI and 11% increase of activity at 1 mM for IsoII Phaseolus vulgaris
3.5.2.5 diethyl dicarbonate 63% increase of activity at 0.1 mM for IsoI and 38% increase of activity at 1 mM for IsoII Phaseolus vulgaris
3.5.2.5 dithiothreitol 20% increase of activity at 1 mM for IsoI and% increase of activity at 0.1 mM for IsoII Phaseolus vulgaris
3.5.2.5 phenyl phosphoramidate 4% increase of activity at 1 mM for IsoI and 10% increase of activity at 1 mM for IsoII Phaseolus vulgaris

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.2.5 4-hydroxymercuribenzoate inactivation, prevented by DTT Phaseolus vulgaris
3.5.2.5 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
3.5.2.5 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
3.5.2.5 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
3.5.2.5 diethyl dicarbonate 89% residual activity at 10 mM for IsoI and 90% residual activity at 10 mM for IsoII Phaseolus vulgaris
3.5.2.5 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
3.5.2.5 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
3.5.2.5 iodoacetic acid 16% residual activity at 5 mM for IsoI and 13% residual activity at 5 mM for IsoII Phaseolus vulgaris
3.5.2.5 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
3.5.2.5 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
3.5.2.5 additional information the enzyme is inhibited by sulfhydryl-reactive agents Phaseolus vulgaris
3.5.2.5 N-ethylmaleimide 54% residual activity at 5 mM for IsoI and 67% residual activity at 5 mM for IsoII Phaseolus vulgaris
3.5.2.5 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
3.5.2.5 p-hydroxymercuribenzoate 10% residual activity at 0.5 mM for IsoI and 15% residual activity at 0.5 mM for IsoII Phaseolus vulgaris
3.5.2.5 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.2.5 additional information
-
 additional information Michaelis-Menten kinetics Phaseolus vulgaris
3.5.2.5 0.059
-
 allantoin pH 7.8, 40°C, isozyme IsoI, different methods Phaseolus vulgaris
3.5.2.5 0.066
-
 allantoin pH 7.8, 40°C, isozyme IsoII, different methods Phaseolus vulgaris

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.2.5 Mn2+
-
 Phaseolus vulgaris
3.5.2.5 additional information metalloenzyme Phaseolus vulgaris

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.2.5 42000
-
 IsoII, native PAGE with acrylamide gradient Phaseolus vulgaris
3.5.2.5 42000
-
 IsoII, SDS-PAGE, after purification two peaks of 45000 and 42000 Da length showing allantoinase activity are obtained Phaseolus vulgaris
3.5.2.5 45000
-
 IsoI, SDS-PAGE, after purification two peaks of 45000 and 42000 Da length showing allantoinase activity are obtained Phaseolus vulgaris
3.5.2.5 53000
-
 IsoI, native PAGE with acrylamide gradient Phaseolus vulgaris

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.2.5 allantoin + H2O Phaseolus vulgaris a key reaction step in the biosynthesis and degradation of ureides, overview allantoate
-
 ?

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.5.2.5 deoxycholate 20% increase of activity in the presence of 0.1% (w/v) deoxycholate Phaseolus vulgaris

Organism

EC Number Organism UniProt Comment Textmining
3.5.2.5 Phaseolus vulgaris
-
-
-
3.5.2.5 Phaseolus vulgaris
-
 cv. Great Northern, two isozymes IsoI and IsoII
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.2.5 DEAE-Sephacel column chromatography, MonoQ HR column chromatography, and Sephacryl S-300 gel filtration Phaseolus vulgaris
3.5.2.5 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

EC Number Source Tissue Comment Organism Textmining
3.5.2.5 fruit highest activity Phaseolus vulgaris
-
3.5.2.5 fruit developing, high enzyme expression level Phaseolus vulgaris
-
3.5.2.5 leaf
-
 Phaseolus vulgaris
-
3.5.2.5 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
-
3.5.2.5 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
-
3.5.2.5 root
-
 Phaseolus vulgaris
-
3.5.2.5 root the amount of IsoII is very low in roots Phaseolus vulgaris
-
3.5.2.5 seedling
-
 Phaseolus vulgaris
-
3.5.2.5 shoot
-
 Phaseolus vulgaris
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.2.5 additional information
-
 activity of isozymes in different bean tissues and during development, overview Phaseolus vulgaris
3.5.2.5 2
-
 crude extract Phaseolus vulgaris
3.5.2.5 295
-
 purified isozyme IsoII Phaseolus vulgaris
3.5.2.5 400
-
 after 200fold purification Phaseolus vulgaris
3.5.2.5 563
-
 purified isozyme IsoI Phaseolus vulgaris

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.5 (S)-allantoin + H2O
-
 Phaseolus vulgaris allantoate
-
 ?
3.5.2.5 allantoin + H2O
-
 Phaseolus vulgaris allantoate
-
 ?
3.5.2.5 allantoin + H2O a key reaction step in the biosynthesis and degradation of ureides, overview Phaseolus vulgaris allantoate
-
 ?
3.5.2.5 additional information no activity with hydantoin and 5-bromouracil Phaseolus vulgaris ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.2.5 monomer
-
 Phaseolus vulgaris
3.5.2.5 monomer 1 * 42000, isozyme IsoII, SDS-PAGE, 1 * 45000, isozyme IsoI, SDS-PAGE Phaseolus vulgaris

Synonyms

EC Number Synonyms Comment Organism
3.5.2.5 allantoin amidohydrolase
-
 Phaseolus vulgaris
3.5.2.5 IsoI 45000 Da enzyme Phaseolus vulgaris
3.5.2.5 IsoII 42000 Da enzyme Phaseolus vulgaris

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.2.5 60
-
-
 Phaseolus vulgaris

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.2.5 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.2.5 201.7
-
 allantoin pH 7.8, 40°C, isozyme IsoII, different methods Phaseolus vulgaris
3.5.2.5 420
-
 allantoin pH 7.8, 40°C, isozyme IsoI, different methods Phaseolus vulgaris

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.2.5 7.7
-
 different methods Phaseolus vulgaris
3.5.2.5 7.8
-
 different methods Phaseolus vulgaris
3.5.2.5 7.8
-
 IsoI Phaseolus vulgaris
3.5.2.5 8.1
-
 IsoII Phaseolus vulgaris

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.2.5 6 8
-
 Phaseolus vulgaris