Christopher Mutsvangwa told a press conference the "game was up" for Mugabe and his wife Grace.
He called for a huge turnout at street protests planned for tomorrow in Harare.
"The generals have done a fantastic job. It's done, it's finished," he said.
"We want to restore our pride and tomorrow is the day... we can finish the job which the army started.
"There's no going back about Mugabe. He must leave."
Mugabe attended a university graduation ceremony today, making a defiant first public appearance since the military takeover that appeared to signal the end of his 37-year reign.
The generals took over late on Tuesday after vice president Emmerson Mnangagwa was abruptly sacked and Mugabe's wife Grace emerged in prime position to succeed her increasingly frail husband.
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