20th Century Fox had earlier suspended the production on the film titled, "Bohemian Rhapsody", due to Singer's "unexpected unavailability".
The film stars Rami Malek as the legendary singer and shooting was underway in London.
"Bryan Singer is no longer the director of Bohemian Rhapsody," the studio said in a statement, according to The Hollywood Reporter.
However, the studio has not named Singer's replacement as yet.
Singer had taken a leave citing "personal health matter concerning Bryan and his family" leading to the studio suspending production.
"Twentieth Century Fox Film has temporarily halted production on 'Bohemian Rhapsody' due to the unexpected unavailability of Bryan Singer," the studio had said.
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