Romney and Holyfield, wearing full protective gear, will spar in the boxing ring for a lighthearted fight before the night's three fights by professional boxers.
The black-tie event is raising money for the Utah-based organization CharityVision, which helps doctors in developing countries perform surgeries to restore vision in people with curable blindness.
Romney's son Josh Romney, who lives in Utah, serves as a volunteer president for CharityVision.
Corporate sponsorships for the event range from $25,000 to $250,000. Organizers say the event is expected to raise $1,000,000.
Ahead of the fight, Romney and Holyfield participated in an official weigh-in Thursday night.
Romney, 68, weighed in at 179 pounds and Holyfield, 52, weighed in at 236 pounds.
In a web video previewing the event, Romney and Holyfield hype the event while working out and wearing boxing shorts.
"It's true, I don't have much of a right hook, but when I get somebody's ear, I can be pretty formidable," Romney said.
"Mitt you can't run, you can't hide. Come get your whoopin'," Holyfield said.
Romney, the most-high profile Mormon in America, is hugely popular in the state, where more than 60 per cent of the residents are members of The Church of Jesus Christ of Latter-day Saints.
Beyond his religious connections, the former Massachusetts governor is remembered by many for turning around Salt Lake City's 2002 Winter Olympics after a bribery scandal.
Romney has recently built a home in the Salt Lake City area and registered as a Utah voter.Suggest a correction