He was the eldest son of Bernhard I, Prince of Anhalt-Bernburg, by his wife Princess Sophie, daughter of King Abel of Denmark.
After the death of his father in 1287, John succeeded him as ruler of the Anhalt-Bernburg, but he had to rule jointly with his third brother, Bernhard II.
Of the two youngest brothers, Henry also was ordained as priest and renounced his rights, whereas Rudolf was possibly dead by the time of his father's death.
John died unmarried and childless after four years of rule.
He was succeeded by his brother Bernhard II, who became sole ruler.