Shakespeare at The George hotel in Huntingdon first began in 1959.
Since then, the production company has put on 563 open air performances in front of over 75,000 people.
But this summer's production of Hamlet will be the last at The George.
Actors say the owner, Greene King, has terminated their annual residency contract.
The group is now seeking alternative venues in the Huntingdon area and say they welcome any offers.