The incident, which happened on the A921 Kinghorn Road, involved one vehicle, a silver Volkswagon Polo, which struck the railway bridge at around 7am today (Wednesday, July 18).
A 49-year-old man was taken to Victoria Hospital by the Scottish Ambulance Service where he sadly died.
The road is currently closed at the junction with the B923 to allow police to investigate and the public are advised to avoid the area, where possible.
Stagecoach bus services have been diverted and service 7 diverting via B9157 & A909, Kinghorn and Burntisland, will not be served as a result of the accident.
A Police Scotland spokesperson said: “Police in Fife are currently in attendance on the A921, Kinghorn Road near Kirkcaldy following a serious road traffic collision that took place at around 7am on Wednesday 18 July.
“The collision involved a Volkswagen Polo car and emergency services are currently at the scene.
“The road is currently closed at the junction with the B923 and the public are advised to avoid the area, where possible.”
Anyone who witnessed the collision or has any information to help with ongoing inquiries is asked to contact 101, quoting incident number 0623 of July 18.