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Literature summary extracted from

  • Arivett, B.; Farone, M.; Masiragani, R.; Burden, A.; Judge, S.; Osinloye, A.; Minici, C.; Degano, M.; Robinson, M.; Kline, P.
    Characterization of inosine-uridine nucleoside hydrolase (RihC) from Escherichia coli (2014), Biochim. Biophys. Acta, 1844, 656-662.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.2.3 gene rihC, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS E. Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.2.3 0.408
-
 uridine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 0.416
-
 adenosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 0.421
-
 ribothymidine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 0.422
-
 Inosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 0.454
-
 Xanthosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 0.682
-
 cytidine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.2.3 33642
-
 2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 33642, His-tagged enzyme, sequence calculation Escherichia coli
3.2.2.3 36000
-
 2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 33642, His-tagged enzyme, sequence calculation Escherichia coli
3.2.2.3 69000
-
 recombinant His-tagged enzyme, gel filtration Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
3.2.2.3 Escherichia coli
-
 gene rihC
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.2.3 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS E. by nickel affinity chromatography, ultrafiltration, dialysis, and anion exchange chromatography Escherichia coli

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.2.3 uridine + H2O = D-ribose + uracil catalytic mechanism with catalytic triad consisting of Tyr223, Tyr227, and His239, overview Escherichia coli
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.2.3 adenosine + H2O
-
 Escherichia coli D-ribose + adenine
-
 ?
3.2.2.3 cytidine + H2O
-
 Escherichia coli D-ribose + cytosine
-
 ?
3.2.2.3 inosine + H2O
-
 Escherichia coli D-ribose + hypoxanthine
-
 ?
3.2.2.3 additional information substrate specificity, overview. No activity with 2'-OH and 3'-OH deoxynucleosides. Nucleosides in which additional groups are added to the exocyclic N6 amino group also exhibit no activity. Nucleosides lacking the 5'-OH group or with the 2'-OH group in the arabino configuration exhibit greatly reduced activity. Purine nucleosides and pyrimidine nucleosides in which the N7 or N3 nitrogens respectively are replaced with carbon also have no activity Escherichia coli ?
-
 ?
3.2.2.3 ribothymidine + H2O
-
 Escherichia coli D-ribose + ?
-
 ?
3.2.2.3 uridine + H2O best substrate Escherichia coli D-ribose + uracil
-
 ?
3.2.2.3 xanthosine + H2O
-
 Escherichia coli D-ribose + xanthine
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.2.3 homodimer 2 * 36000, recombinant His-tagged enzyme, SDS-PAGE, 2 * 33642, His-tagged enzyme, sequence calculation Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
3.2.2.3 inosine-uridine nucleoside hydrolase
-
 Escherichia coli
3.2.2.3 RihC
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.2.3 32
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.2.3 1.12
-
 cytidine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 1.15
-
 adenosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 1.29
-
 ribothymidine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 4.31
-
 Inosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 6.3
-
 Xanthosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 10.85
-
 uridine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.2.3 7.5
-
-
 Escherichia coli

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.2.3 5 9 pH profile is bell-shaped with the enzyme showing significant activity over a wide pH range, 50% activity at pH 5.0 and pH 9.0 Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.2.3 2.8
-
 adenosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 10.2
-
 Inosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 13
-
 Xanthosine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 16
-
 cytidine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 26
-
 uridine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli
3.2.2.3 31
-
 ribothymidine pH 7.2, 32°C, recombinant His-tagged enzyme Escherichia coli