Kelham Island Brewery opened in 1990 in the beer garden of the Fat Cat in Alma Sreet, and at the time was the city's first independent brewery in almost a century. The original site was later converted to a visitor centre and a new brewhouse opened in 1999.
By the end of the 1990s Sheffield's other main independent brewers had all shut, leaving Kelham Island as the oldest.
The owners posted on their Facebook page: "It is with deep sadness that we are having to announce that we have brewed our final batch of Pale Rider and the brewery will shortly be closing.
"We would like to thank everybody who supported us over the last 32 years - especially our staff and customers.
"We have brewery tours scheduled for Saturday 7 and Saturday 14 May, these will be going ahead as planned, but no further tours will be booked in. If you have a brewery tour voucher with a code on the back, these can now be redeemed up to a value of £25 against any item on the shop on our website.
"The Fat Cat will remain open, with Kelham Island Brewery beer available on cask and in bottles/cans while stocks last.
"It is our intention that all creditors will be repaid in full and this includes any customers that have vouchers they do not wish to redeem at the shop. Should you have any queries in this regard, please contact Begbies Traynor using the following e-mail address:- [email protected] Furthermore, any interest in any of the assets should also be directed to this email address."
The early operation produced 10 barrels per week, but by 1999 capacity increased to 50.
By 2008, the brewery expanded again and the business ended with a weekly output of 100 barrels.