Javier Cabello takes the reins for the short-term just as he did in the brief period after Argentine Luis Zubeldia was dismissed in September.
"The blue-and-whites have decided to terminate Gianni De Biasi's contract after the latest La Liga results," Alaves said in a statement.
"For the moment Javier Cabello will be handed the leadership of the squad."
Former Albania coach De Biasi, 61, suffered six defeats in eight league games in charge but also earned the club their only top-flight victories this season, against Levante and Espanyol.
(Reporting by Rik Sharma, editing by Ed Osmond)
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