HMV in Gloucester will close within the next six weeks.
The national music, film and games retailer fell into administration in January but there had been high hopes the store in King's Walk shopping centre would be saved.
However administrators Deloitte announced it was being axed yesterday along with the Cheltenham branch and 35 other stores across the country. The closure will leave another major gap in King's Walk, which has already lost the likes of Jessops, which was next door, and Shakeaway.
No-one was available from the shopping centre's management yesterday to comment on the imminent closure.
No fixed date is set for the closure of the stores which will continue to trade in the meantime. But it is expected closures will take place over the next four to six weeks in parallel with the 66 stores that are already scheduled to close.
Nick Edwards, joint administrator at Deloitte, said: "As part of our ongoing review of HMV's financial position, we have undertaken a further review of the store portfolio and have identified stores for closure.
"This step has been taken to enhance the prospects of the restructured business continuing as a going concern.
"We are extremely grateful to the staff for their continued strong support and commitment during an understandably difficult period.
"Suppliers and landlords remain supportive and we appreciate their ongoing assistance."
The affected stores employ 464 staff.