Little Chef is to close 67 of its 161 outlets in a move expected to lead to between 500 and 600 job cuts.
The sites earmarked for closure have been trading unprofitably for a number of years, the company said.
The motorway restaurant group blamed the weak economy and the location of the branches for their poor performance.
The job losses will be a mixture of full and part-time positions. About 1,500 staff will remain.
Little Chef was saved by turnaround group RCapital when it fell into administration in 2007. The following year several branches were revamped by Michelin-starred chef Heston Blumenthal as part of a Channel 4 show. In a statement, the firm said the decision to close a number of its sites was not taken lightly, but was "essential" to allow the company to develop its remaining sites.
The company has had to respond to competition from supermarkets and coffee chains at service stations.
But unions have criticised its current owners for failing to protect jobs.
"This is dreadful news for the employees in these restaurants due for closure," said GMB senior officer Paul Maloney.