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Literature summary for 1.14.13.212 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 PubMed
Search for more literature for 1.14.13.212

Cloned(Commentary)

Cloned (Comment) Organism
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. I7ASS7
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,3,7-trimethylurate + NADH + H+ + O2
-
 Pseudomonas sp. 1,3,7-trimethyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
1,3-dimethylurate + NADH + H+ + O2
-
 Pseudomonas sp. 1,3-dimethyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
1-methylurate + NADH + H+ + O2 low activity Pseudomonas sp. 1-methyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
3,7-dimethylurate + NADH + H+ + O2 best substrate Pseudomonas sp. 3,7-dimethyl-5-hydroxyisourate + NAD+ + H2O
-
 ?
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. ?
-
 ?

Synonyms

Synonyms Comment Organism
NADH-dependent trimethyluric acid monooxygenase
-
 Pseudomonas sp.
tmuM
-
 Pseudomonas sp.

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

Cofactor

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

General Information

General Information Comment Organism
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.
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.
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]

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