Tiago Klimeck, 27, was enacting the suicide of Judas during the performance on Good Friday in the city of Itarare.
The actor was hanging for four minutes before fellow performers realized something was wrong.
Tiago Klimeck was taken to hospital suffering from cerebral hypoxia but died on Sunday.
The Passion play was being performed in Itarare, 345 km (214 miles) west of Sao Paulo.
Tiago Klimeck was re-enacting the scene in which Judas commits suicide in repentance for his betrayal of Jesus Christ.
Police are investigating the apparatus that was meant to support Tiago Klimeck. It appears the knot may have been wrongly tied.
When the actors realized something was wrong, Tiago Klimeck was taken down and found to be unconscious.
The Santa Casa de Itapeva hospital has confirmed the death and a post-mortem examination took place yesterday.
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