E-bikes will be available to hire in Stirling, Falkirk and Clackmannanshire, with 12 charging stations across the council areas.
The £500,000 scheme, run by the Forth Environment Link charity and funded by Transport Scotland, NHS Forth Valley, Sestran and the local authorities, will enable people to hire a bike for £2 per journey or a £65 annual fee for unlimited journeys.
Launching the project at Forth Valley Royal Hospital, Public Health Minister Joe FitzPatrick said: "I'm delighted to join Forth Environment Link and NHS Forth Valley to launch Forth Bike - Scotland's largest electric bike hire scheme.
"This flexible scheme provides affordable and easy access to modern e-bikes at a fraction of the cost required to buy the latest models.
"This progressive initiative, supported by a range of partners and funded by the Scottish Government, will widen access to the benefits of active travel and increase opportunities for sustainable transport over single-occupancy car journeys for everyday trips."
He added: "This initiative supports our climate ambitions, improves air quality and benefits our health and sense of well-being - directly contributing to our objective of developing an active nation where more people can enjoy the benefits of walking and cycling than ever before."
Forth Environment Link executive director Clara Walker said: "Forth Bike aims to connect communities across Forth Valley, opening up access to services and employment opportunities by offering a flexible, inexpensive and environmentally friendly way to travel.
"Cycling is also great for our health. E-bikes give as little or as much assistance as you need, making them great for those getting back in the saddle or new to exercise."
The e-bikes will last for up to 60 miles on a single charge and councillors from Clackmannanshire, Falkirk and Stirling said they hope the scheme will encourage locals and tourists to opt for the bikes over cars.
Falkirk Council environment spokesman Paul Garner said: "This exciting project will deliver low carbon travel options for residents and visitors to explore the area.
"The bike share scheme will significantly add to the facilities for tourism and the town centre.
"We are hopeful that more cycle hire base units will be installed in other sites soon."
Stirling Council environment and housing committee convener Jim Thomson said: "Electric bikes will give residents and tourists the freedom and benefits of cycling, with a little extra assistance.
"By providing the opportunity for people to use electric bicycles in a range of locations throughout Stirling, it is not only good for a person's individual health and well-being, it is also positive for the environment."