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The two rivals were rumoured to be in talks over locking horns one more time but an agreement was not reached and instead Tyson fought Roy Jones Jr in a eight-round exhibition that was declared a draw by the unofficial judges at the Staples Center.
Holyfield, who last fought in 2011, has gone on record to say he would not fight his former opponent again for anything less than 25 million dollars.
But in a new statement, he has urged Tyson, the man who infamously bit his ear in 1997, to sign the contract and make the bout happen.
The Real Deal, 58, said: "My side tried to make the fight happen and we got nothing but excuses. Now I can see why he wanted a tune-up fight before thinking about me.
"Roy Jones was a good local opponent for Mike.
"But a fight with me would be a global event and the only fight that anyone wants to see fight is between us.
"There is absolutely no reason why we shouldn't make it happen.
"No more excuses. This is the fight that must happen for both our legacies. Saturday night you said you were ready to fight me, so sign the contract and get in the ring, Tyson.
"The world is waiting and it's on you. I'm ready."
Tyson himself has already expressed an interest in meeting Holyfield for a third time having lost both fights against the five-time world champion.
"Maybe Evander needs to talk to me because every time my business associates talk to his business associates, it doesn't turn out well," Tyson said after the Jones Jr fight.
"So I don't know. If you could see what we made tonight, if these guys really care about the welfare of Evander they would have had this fight with Evander.
"Maybe we'll do another but whoever's he's with who's handling him is totally wrong."
Holyfield is nearing 60 years of age but remains in superb shape, having posted regular videos of him training in the gym.
He's supposedly fighting Glenn McCrory for Bare Knuckle Boxing in April - even though the two are expected to be wearing gloves.
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