Tiago Klimeck, 27, was enacting the suicide of Judas during the performance on Good Friday in the city of Itarare.
While acting the scene in which Judas commits suicide after betraying Jesus, he was supposed to ‘hang himself’ from a rope then pretend to be dead.
But the safety vest the actor was using under his robe is believed to have risen up around his neck, strangling him.
The actor was hanging for four minutes before fellow performers realized something was wrong.
Klimeck was taken to hospital suffering from cerebral hypoxia but died on Sunday.
The Passion play was being performed in Itarare, 345km (214 miles) west of Sao Paulo.
Police are investigating the apparatus that was meant to support Klimeck. It appears the knot may have been erroneously tied.
When the actors realized something was wrong, Klimeck was taken down and found to be unconscious.
The Santa Casa de Itapeva hospital has confirmed the death and an autopsy will take place on Monday.