Sheffield Chamber of Commerce is urging the Government to resolve quickly concerns raised by businesses in the Lower Don Valley affected by the proposed route of high speed trains.
Current plans pose a threat to manufacturing companies.
Chamber executive director Richard Wright said: “It is clearly unacceptable to choose a route that cuts through major businesses. What we need is an early resolution to the consultation to remove the doubt and allow the businesses along the route to get on and operate and invest with confidence.”
Consultation on the proposed route is underway.