The former WWE and UFC champion pinned the Deadman -- real name Mark William Calaway -- to the ground with his signature F-5 achieving what 21 other wrestlers couldn't.
"The STREAK is OVER. @BrockLesnar DEFEATS #Undertaker!
And we never thought we'd tweet those words. #21and1" Wrestlemania tweeted after the match.
The event's result left the New Orleans audience in awe and reportedly shouting "no" and then "yes" once the referee counted 1-2-3 on the Taker's shoulder.
While Lesnar won the fight 21-1, The Undertaker -- who is the only remaining active competitor from 1993's WWE Raw -- still got a standing ovation from spectators at the Superdome.
Sunday's match results have led many to believe that the fight will be Undertaker's last in WWE. Bleacher Report pointed to the Undertaker's age -- he just turned 49 -- as another factor in the company's decision to end his run with them.