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Literature summary extracted from

  • Muratsubaki, H.; Enomoto, K.; Soejima, A.; Satake, K.
    An enzyme cycling method for measurement of allantoin in human serum (2008), Anal. Biochem., 378, 65-70.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.1.47 analysis use of glucose dehydrogenase in enzyme cycling method for measurement of allantoin in human serum Priestia megaterium
3.5.3.9 analysis development of an enzyme cycling method for measuring allantoin concentrations in human serum Metabacillus fastidiosus
6.3.1.5 analysis method for measurement of allantoin in human serum. Serum allantoin is converted to allantoate by the action of allantoinase,and endogenous ammonia is simultaneously removed by the action of glutamine synthetase II. In the second step, L-methionine sulfoximine is used to inhibit glutamine synthetase II, and ammonia is liberated from allantoate by the activity of allantoate amidohydrolase. The ammonia is then converted to NAD by NAD synthetase. Subsequent action of glucose dehydrogenase and diaphorase acts to cycle the formed NAD between its oxidized and reduced forms, resulting in the production of WST-1 formazan, which is monitored at 450 nm. The assay standard curve is linear from 0 to 70 microM allantoin Geobacillus stearothermophilus

General Stability

EC Number General Stability Organism
3.5.3.9 preparation of allantoate amidohydrolase stabilized by the addition of 0.1 mM EDTA Metabacillus fastidiosus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.3.9 Mn2+ essential for enzyme activity, catalyzes the second step of the developed assay, ammonia removal reaction in the presence of Metabacillus fastidiosus

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 Glycine max
-
 allantoate
-
 ?
3.5.3.9 allantoate + H2O Metabacillus fastidiosus
-
 ureidoglycine + NH3 + CO2
-
 ?
3.5.3.9 allantoate + H2O Metabacillus fastidiosus ATCC 29604
-
 ureidoglycine + NH3 + CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.47 Priestia megaterium
-
 commercial preparation
-
3.5.2.5 Glycine max
-
-
-
3.5.3.9 Metabacillus fastidiosus
-
 ATCC29604
-
3.5.3.9 Metabacillus fastidiosus ATCC 29604
-
 ATCC29604
-
6.3.1.5 Geobacillus stearothermophilus
-
 commercial preparation
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.2.5
-
 Glycine max
3.5.2.5 native enzyme from seeds Glycine max

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.5.2.5 seed
-
 Glycine max
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.3.9 additional information
-
 enzyme cycling method for measuring allantoin concentrations in human serum, ammonia is liberated from allantoate by the activity of allantoate amidohydrolase, 70 microM allantoin and a concentration of 2 U/ml allantoate amidohydrolase required for maximum absorbance, measurement of allantoin levels in healthy subjects Metabacillus fastidiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.5 (S)-allantoin + H2O
-
 Glycine max allantoate
-
 ?
3.5.2.5 allantoin + H2O
-
 Glycine max allantoate
-
 ?
3.5.2.5 allantoin + H2O development of a enzyme cycling method for measuring allantoin concentrations in human serum involving allantoinase, glutamine synthetase II, EC 6.3.1.2, allantoate amidohydrolase, EC 3.5.3.9, and NAD synthetase, EC 6.3.1.5, followed by action of glucose dehydrogenase, EC 1.1.1.47, and diaphorase, EC 1.6.99.2, in the presence of glucose and 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, overview, optimal reaction for allantoinase at 0.07 mM allantoin concentration Glycine max allantoate
-
 ?
3.5.3.9 allantoate + H2O
-
 Metabacillus fastidiosus ureidoglycine + NH3 + CO2
-
 ?
3.5.3.9 allantoate + H2O
-
 Metabacillus fastidiosus ATCC 29604 ureidoglycine + NH3 + CO2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.3.9 allantoate amidohydrolase
-
 Metabacillus fastidiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.3.9 37
-
 assay at Metabacillus fastidiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.3.9 7.3
-
 assay at Metabacillus fastidiosus