Exodeoxyribonuclease I (EC 3.1.11.1, Escherichia coli exonuclease I, E. coli exonuclease I, exonuclease I) is an enzyme that catalyses the following chemical reaction:

Exonucleolytic cleavage in the 3′- to 5′-direction to yield nucleoside 5′-phosphates

Preference for single-stranded DNA. The Escherichia coli enzyme hydrolyses glucosylated DNA.

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