Poundland in Hitchin town centre will permanently close on July 16, the Comet can reveal.
A spokesperson for the discount store chain said the company had not been able to secure a long-term lease for the unit in Bancroft.
He said: "We’ve sadly decided to close our Hitchin store on July 16.
"I'm afraid we've been unable to secure a long-term lease that would enable us to keep trading.
"We know how disappointing this will be to customers and colleagues."
Asked if the closure will result in redundancies, the Poundland spokesperson said: "Whenever we have to close a store, we do all we can to look for other opportunities for colleagues, and that work is now under way."
Poundland moved into the former Wilko unit in Bancroft in October last year, after Wilko went into administration.
Poundland owner Pepco had agreed to purchase 71 Wilko stores, including the Hitchin branch, following the collapse of the high street chain.
Poundland previously had a unit in Hitchin's High Street.
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