Peter and Isobel Doherty were at a bus stop during a holiday in Orlando when a Ford Focus hit them.
Mr Doherty, 68, was left in a coma but died on Sunday. Mrs Doherty suffered broken ribs, bruising and cuts.
A Foreign Office spokesman said: “We can confirm a British national died in hospital on 23 September.
“We are continuing to provide consular assistance to the family.”
The couple, from Dunbartonshire, were waiting for a bus outside Millennia Mall on 16 September when the accident happened.
Relatives of the couple are believed to have flown to the United States. Mr Doherty was the owner of fencing company Doherty & Lafferty.