Orf135 from Escherichia coli Is a Nudix Hydrolase Specific for CTP, dCTP, and 5-Methyl-dCTP

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To characterize the enzymatic properties of Orf135, a Nudix hydrolase from Escherichia coli, specific for CTP, dCTP, and 5-methyl-dCTP.
Cloning and overexpression of the Orf135 gene.
Purification and characterization of the Orf135 protein from Escherichia coli.
Enzymatic activity assays to determine substrate specificity and kinetics.
Orf135 exhibits highest activity on 5-methyl-dCTP (k cat/K m = 301,000 m−1s−1).
Demonstrated lower activities for CTP (k cat/K m = 47,000 m−1 s−1) and dCTP (k cat/K m = 18,000m−1 s−1).
Orf135 requires magnesium ions for optimal activity and operates under alkaline pH conditions.

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Examines Orf135's enzyme activity on CTP, dCTP, and 5-methyl-dCTP, suggesting its role in pyrimidine metabolism.

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O’Handley et al. (2001) studied this question.

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