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Literature summary for 3.2.2.3 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)

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

KM Value [mM]

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
gene rihC
-

Purification (Commentary)

Purification (Comment) Organism
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

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adenosine + H2O
-
Escherichia coli D-ribose + adenine
-
?
cytidine + H2O
-
Escherichia coli D-ribose + cytosine
-
?
inosine + H2O
-
Escherichia coli D-ribose + hypoxanthine
-
?
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 ?
-
?
ribothymidine + H2O
-
Escherichia coli D-ribose + ?
-
?
uridine + H2O best substrate Escherichia coli D-ribose + uracil
-
?
xanthosine + H2O
-
Escherichia coli D-ribose + xanthine
-
?

Subunits

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

Synonyms

Synonyms Comment Organism
inosine-uridine nucleoside hydrolase
-
Escherichia coli
RihC
-
Escherichia coli

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
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]

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