St Clement's Church, on Star Lane, has been closed since 1979 but is being transformed into Ipswich Arts Centre.
After two years of planning, the first night of live music took place as part of SPILL Festival of Performance.
Carol Gant, the centre's director, said it was "wonderful to hear the church filled with live music and singing again after so many years".
Dead Rat Orchestra, She Shanties, Cath & Phil Tyler, Documents and MacGillvray were on the line-up at the former church, which was built in the 15th Century.
Mrs Gant said about 200 people attended the gig.
"The audience and performers were so impressed by the way the voices echoed around the space. It was a wonderful night," she said.
"After 30 years of disuse" it was "a great moment for the town", Cad Taylor, part of the community interest company behind the centre, added.
Despite recently receiving permission to change the use of the church into a venue, the group is still fundraising to complete work at the centre.
It is currently asking people to back a bid for Â£25,000 from Aviva.
More than 30 years of silence ended at a Suffolk church on Friday, as it hosted its first gig as an arts centre.