The 63-year-old woman was cycling on Bridge Street, St Andrews, Fife, at around 3.50pm on Saturday when she collided with a man on foot.
The pair both fell on to the road near to the junction with Kinnessburn Road, resulting in the cyclist suffering serious head injuries. She was taken to Ninewells Hospital in Dundee, where she later died.
Road policing officers are asking anyone who can assist with their inquiries to come forward.
Sergeant Iain Hutchison, said they were eager to hear from members of the public who were in the area at the time and particularly an elderly couple and a male pedestrian who were walking down Bridge Street at the time of the accident.