The transformation of Interstate 4 could take five years and more than $2 billion to complete.
More than $250 million are up for grabs for small or disadvantaged businesses.
Express toll lanes are planned, more than 50 new bridges are to be built for the project and all exits will be on the right.
A workshop Thursday bridged the gap between businesses and the multimillion-dollar opportunity.
Ann McNeill of MCO Construction spent the morning trying to convince four teams to hire her construction firm for the I-4 project.
"Because of the economy a few years ago, this is a major boost to all firms; small, medium and large firms," McNeil said.
One of those four teams will ultimately be chosen for the project.
Dee Carlson of Arcpoint Labs went from room to room selling her 2-year-old Altamonte Springs company.
She's offering to do onsite drug testing of construction workers and said her new business needs the work.
"I think it would be increasingly difficult to get your word out to as many employers as you can reach in this type of function," Carlson said.
More than 250 businesses from around the state have signed up to do interviews over the next two days.
"To have an opportunity to be a part of the process and this is the first time in a long time that we've had that opportunity," McNeil said.
The Florida Department of Transportation will announce the team chosen to work on the I-4 project early next year.