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Literature summary extracted from

  • Mohanty, S.K.; Yu, C.L.; Das, S.; Louie, T.M.; Gakhar, L.; Subramanian, M.
    Delineation of the caffeine C-8 oxidation pathway in Pseudomonas sp. strain CBB1 via characterization of a new trimethyluric acid monooxygenase and genes involved in trimethyluric acid metabolism (2012), J. Bacteriol., 194, 3872-3882.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.212 gene tmuM, DNA and amino acid sequence determination and analysis, genes involved in the caffeine C-8 oxidation pathway are located in a 25.2-kb genomic DNA fragment of CBB1, including cdhABC (coding for caffeine dehydrogenase) and tmuM (coding for TmuM) Pseudomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.212 0.0012
-
 1-Methylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 0.0013
-
 3,7-dimethylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 0.0102
-
 1,3,7-trimethylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 0.1265
-
 1,3-dimethylurate pH 7.5, 30°C Pseudomonas sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.17.5.2 20000
-
 gamma subunit Pseudomonas sp.
1.17.5.2 32000
-
 beta subunit Pseudomonas sp.
1.17.5.2 90000
-
 alpha subunit Pseudomonas sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.212 1,3,7-trimethylurate + NADH + H+ + O2 Pseudomonas sp.
-
 1,3,7-trimethyl-5-hydroxyisourate + NAD+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.212 Pseudomonas sp. I7ASS7
-
-
1.17.5.2 Pseudomonas sp.
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.212 native enzyme 55fold from strain CBB1 by anion exchange, affinity, and hydrophobic interaction chromatography, followed by gel filtration Pseudomonas sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.212 4.128
-
 purified native enzyme, pH 7.5, 30°C Pseudomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.212 1,3,7-trimethylurate + NADH + H+ + O2
-
 Pseudomonas sp. 1,3,7-trimethyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
1.14.13.212 1,3-dimethylurate + NADH + H+ + O2
-
 Pseudomonas sp. 1,3-dimethyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
1.14.13.212 1-methylurate + NADH + H+ + O2 low activity Pseudomonas sp. 1-methyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
1.14.13.212 3,7-dimethylurate + NADH + H+ + O2 best substrate Pseudomonas sp. 3,7-dimethyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
1.14.13.212 additional information enzyme TmuM prefers trimethyluric acids and, to a lesser extent, dimethyluric acids as substrates, but it exhibits no activity on uric acid Pseudomonas sp. ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.17.5.2 trimer 1 * 90000 + 1 * 32000 1 * 20000, alpha, beta, and gamma subunits Pseudomonas sp.

Synonyms

EC Number Synonyms Comment Organism
1.14.13.212 NADH-dependent trimethyluric acid monooxygenase
-
 Pseudomonas sp.
1.14.13.212 tmuM
-
 Pseudomonas sp.
1.17.5.2 caffeine dehydrogenase
-
 Pseudomonas sp.
1.17.5.2 Cdh
-
 Pseudomonas sp.
1.17.5.2 cdhABC
-
 Pseudomonas sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.13.212 30
-
 assay at Pseudomonas sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.212 0.485
-
 1-Methylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 1.97
-
 3,7-dimethylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 3.08
-
 1,3-dimethylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 7.48
-
 1,3,7-trimethylurate pH 7.5, 30°C Pseudomonas sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.212 7.5
-
 assay at Pseudomonas sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.212 FAD flavoprotein Pseudomonas sp.
1.14.13.212 NADH dependent on Pseudomonas sp.

General Information

EC Number General Information Comment Organism
1.14.13.212 evolution genes involved in the caffeine C-8 oxidation pathway are located in a 25.2-kb genomic DNA fragment of CBB1, including cdhABC (coding for caffeine dehydrogenase) and tmuM (coding for TmuM), Physical map of genes for caffeine transformation in a 25.2-kb gene cluster in Pseudomonas sp. strain CBB1, overview Pseudomonas sp.
1.14.13.212 metabolism the enzyme takes part in the caffeine C-8 oxidation pathway as the second enzyme, a NADH-dependent trimethyluric acid monooxygenase that catalyzes the conversion of trimethylurate to 1,3,7-trimethyl-5-hydroxyisourate. This product spontaneously decomposes to racemic 3,6,8-trimethylallantoin Pseudomonas sp.
1.14.13.212 additional information homology models of trimethyluric acid monooxygenase against uric acid oxidase HpxO (which catalyzes uric acid to 5-hydroxyisourate) reveal a much bigger and hydrophobic cavity to accommodate the larger substrates Pseudomonas sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.212 25
-
 1,3-dimethylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 408.3
-
 1-Methylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 735
-
 1,3,7-trimethylurate pH 7.5, 30°C Pseudomonas sp.
1.14.13.212 1490
-
 3,7-dimethylurate pH 7.5, 30°C Pseudomonas sp.