DeSantis, appearing a bill signing ceremony in St. Petersburg, said the order is meant to bridge the gap before legislation limiting local measures officially comes into effect on July 1.
The legislation, passed by the Florida Legislature last week, bars state and local governments from shuttering businesses and schools during health emergencies and bans "vaccine passports."
"The bill ensures that neither the state or local governments can close businesses or keep kids out of in-person instruction unless they satisfy a demanding and continuous justification," DeSantis told reporters, adding that Florida is no longer in a state of emergency.
DeSantis said the immediate suspension of local COVID-19 restrictions is justified by data showing that vaccines have been effective and that that much of the state's elderly population has already been inoculated.
"I think folks that are saying they need to be policing people at this point, if you're saying that, you're really saying you don't believe in the vaccines, you don't believe in the data, you don't believe in the science," he added. "We've embraced the vaccines."
Florida, which struggled with rising cases last summer, has so far fully vaccinated 37% of its population, according to state health officials. The state has recorded more than 2.2 million cases and 35,000 deaths related to COVID-19 since the start of the pandemic.
DeSantis, who was among the first U.S. governors to allow businesses to reopen following the initial waves of the disease last year, criticized other states for keeping COVID-19 restrictions in place.
"Now in the state of Florida, we worked very hard, particularly since this summer, to jettison those types of policies and we focused on lifting people up," he said.
The governor's move brought negative reactions from some local leaders in Florida.
"To be clear, cities like St. Pete, Tampa, Orlando, Miami and Miami Beach, saved Florida and the governor's behind throughout this pandemic," tweeted St. Petersburg Mayor Rick Kriseman.
"Can you imagine if each city had been led by Ron DeSantis? How many lives would have been lost? What would our economy look like today?"