He tweeted that he "did not make, and do not have, any such recordings", a day before he was required to hand over any such tapes by Congress.
Mr Trump kick-started speculation of recordings in a tweet he posted days after firing Mr Comey in May.
He said: "James Comey better hope there are no 'tapes' of our conversations".
This tweet resulted in a flurry of allegations and speculation that Mr Comey was fired in order to hamper the FBI's investigation into alleged Russian interference in last year's election and possible collusion between the Trump campaign team and Moscow.
Whatever its intention, the tweet ultimately led to the appointment of a special counsel that has taken over the investigation.
It all stems from last year's presidential election and mounting evidence that Russian cyber hackers targeted US electoral systems in order to help Donald Trump win - something Moscow has strongly denied.