The centre-right opposition on Thursday filed a motion of no-confidence in Justice Minister Alfonso Bonafede.
The minister has come under fire over the release of 376 top mafiosi for ill health in recent months.
He has also been criticised over an alleged failure to appoint anti-mafia prosecutor Nino Di Matteo as head of the Italian prison system in 2018.
Bonafede reiterated Thursday that there had never been any let-up in the fight against Italy's mafias.
He has said the government has a measure in the pipeline that would revise the release of mafia bosses and mafiosi for health reasons because of the "changed picture" of the coronavirus emergency.
Bonafede also reiterated Thursday there was "no link" between the Di Matteo issue and that of the released bosses.
National anti-mafia prosecutor Cafiero De Raho said Thursday the release of the mafiosi could have been averted.
"People got carried away by fears of contagion, when thermoscanners would have been enough," he told Italian radio.
Bonafede's 5-Star Movement (M5S) said the centre right's move would man "undermining confidence in the fight against corruption, the fight against the mafias, reforms to make trials shorter and more efficient, and measures to make sure no one goes unpunished."
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