Kirkcaldy's Tesco store is to shut on April 4th, leading to the loss of 189 jobs.
Staff were told the news this morning during a meeting.
Politicians say the decision now raises questions over the future of the Postings Shopping Centre and the town centre's only post office, situated within the Tesco store.
Supermarket bosses say the closure is necessary due to business falling significantly short of where they want it to be.
A total of 43 unprofitable stores are to shut across the UK. 2, 000 positions will go across Britain.
Tesco Chief Executive Dave Lewis said:
“Earlier this month I announced that our performance as a business has fallen significantly short of where we would want it to be and that to protect the future of the business in the UK we would close 43 unprofitable stores.
It is with great sadness that I have to tell you that our Kirkcaldy Superstore is one of the 43 we plan to close. The decision to close the store has been exceptionally difficult to take. I recognise it will affect many hard working colleagues, our customers and the local community.
Our priority is to explain what this announcement means for our colleagues and wherever possible, offer them alternative roles with Tesco. I would like to thank all our customers who have shopped in our store. We will continue to do our best to serve them through our other stores in Fife, as well as through our dotcom service.”