The month-long contest, called the Python Challenge, begins on Saturday.
About 550 people have signed up to compete for two prizes: $1,000 (£620) for the longest python killed and $1,500 for the most pythons taken.
First imported as pets, the feral snakes vie with native animals for food and have devastated mammal populations.
The US banned the import of the carnivorous snakes last year. But some wildlife officials said the ban came too late - after the southern Asia natives had begun breeding in the Everglades.
Burmese pythons, which can grow up to 23ft (7m) long, eat small mammals, birds and alligators.
Hunters from more than 25 states have signed up for the event. Among them will be US Senator Bill Nelson, a 70-year-old Democrat.
The state will train hunters to avoid mistaking the python for native snakes, Florida Fish and Wildlife Conservation Commission (FWC) spokeswoman Carli Segelson said.
The FWC recommends shooting the snakes in the head or using a device that drives a bolt into the brain. Hunters will be able to keep the pelts.
Decapitation is not recommended unless the brain is immediately destroyed, as snakes can remain conscious for a time after the head is separated from the body.
|The largest Burmese python captured in the Everglades was 17ft 7in (5.3m)|