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Born in Orsa, Dalarna, Zetterstéen began his studies at Uppsala University in 1884, became a Ph.D. and docent of Semitic languages in 1895. He was acting professor of Oriental languages at Lund University 1895-1904 and professor of Semitic languages in Uppsala 1904-1931. He also studied under Eduard Sachau at the University of Berlin.
As a semitist, Zetterstéen was foremost an Arabic philologist, but he was also well-oriented in non-Semitic languages such as Persian, Turkish and Nubian. Beside a large number of text editions and studies, he published a Swedish translation of the Qur'an, a number of popular works, and wrote several articles in the Nordisk familjebok.