Poundstretcher on Bridge Street is set to close in December.
However, the company's second shop off Newgate Street will remain open.
Nadir Lalani, Poundstretcher Executive Chairman: “We would like to thank all our loyal customers and the hard-working staff of our Bridge Street store.
“We are currently attempting to transfer staff members following this closure to other stores.
“Whilst there is another store remaining open in the local area at The Mayfair Centre, with a wider selection of branded household products all at bargain prices, we have also opened 50 stores across the UK from April this year, with more stores opening within the coming months.”
The shop will be closing on Saturday, December 8.
Philip Jackson chairman of Worksop Business Forum has urged Worksop residents to support the high street in light of the latest shop closure.
He said: “We are extremely disappointed that Poundstretcher have taken the decision to close their Bridge Street store.
“I have said many times that if the people of Worksop don’t support the high street they risk loosing it.
“We must encourage people to shop local.”
Poundworld which had a shop next to the Poundstretcher on Bridge Street closed in August after the company fell into administration leaving two large shops vacant on the street.