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In 1513 he entered the congregation of the Order of the Augustinian Canons of San Salvatore of Bologna, taking up residence in the monastery of San Secundo, one of the order's houses in Gubbio. In 1524 he went to the mother cloister in Bologna, from where he briefly attended courses in Hebrew and rhetoric at the University of Bologna. In 1529 he was sent by his congregation to the Monastery of Sant' Antonio di Castello in Venice, where, due to his expertise in biblical languages and humanist textual criticism, he was placed in charge of the monastery's library, donated to the canons by Cardinal Domenico Grimani. Many of the collection's biblical, Hebrew, and philosophical works had once been owned by Pico della Mirandola.
Over the next several years (1529–33) Steuco wrote a series of polemical works against Luther and Erasmus, the latter of whom he accused of helping to foment the Protestant Revolt against the Church. These works show Steuco's staunch support of the traditions and practices of the Church, including a strident defense of papal authority. Part of his output during this period included a major set of annotations on the Pentateuch, titled Veteris testamenti ad Hebraicam veritatem recognitio, in which he used Hebrew and Greek manuscripts from the Grimani Library to correct Jerome's Vulgate translation of the Old Testament text. When explaining the text, he never strayed from the literal and historical meaning. An interesting juxtaposition to this work of humanist biblical exegesis was a syncretic philosophical work that he wrote in this period, to which he gave the title Cosmopoeia.
His polemical and exegetical works attracted the notice of Pope Paul III, and in 1538 the pope made Steuco bishop of Chisamo on the island of Crete, and librarian of the papal collection of manuscripts and printed works in the Vatican. While he never visited his bishopric in Crete, Steuco did actively fulfill his role as Vatican Librarian until his death in 1548.
While in Rome he authored Old Testament annotations on the Psalms and Job, again relying heavily upon Hebrew sources to help annotate and correct the texts. Also from this period dates a major work entitled the De perenni philosophia, dedicated to Paul III, in which he attempted to show that many of the ideas expounded by the sages, poets, and philosophers from classical antiquity were in essential harmony with the central tenets of the Catholic faith. This work has a slight polemical edge to it, as Steuco crafted a number of his arguments to lend support to several theological positions that had recently come under question in Italy by reformers and critics of the traditional Catholic faith.
As a Roman humanist, he also took a deep interest in the classical ruins of Rome, and in the urban renewal efforts of Paul III throughout the city. Of particular note in this vein are a series of short orations that he wrote and possibly delivered at the papal court, urging Paul III to refurbish the aqueduct known as the Aqua Virgo, in order to supply Rome with adequate fresh water, and as a major key to the revitalization of the city itself.
In 1547 Steuco was sent by Paul III to attend the Council of Trent, where he could be counted upon to uphold papal prerogatives and authority. He died in 1548 while in Venice on break from the Council. He is now buried in Gubbio.
- Freudengerger, Theobald. Augustinus Steuchus aus Gubbio, Augustinerchorherr und papstlicher Bibliothekar (1497-1548). Munster in Westfalen: Aschendorffsche, 1935
- Delph, Ronald K. "From Venetian Visitor to Curial Humanist: The Development of Agostino Steuco's ‘Counter’-Reformation Thought." Renaissance Quarterly 47 (1994): 102-39.
- Delph, Ronald K. "Renovatio, Reformatio, and Humanist Ambition in Rome." In Heresy, Culture and Religion in Early Modern Religion. Edited by Ronald K. Delph, Michelle M.Fontaine, and John Jeffries Martin, pp. 73–92. Kirksville, Mo: Truman State University Press, 2006.
- Herbermann, Charles, ed. (1913). "Agostino Steuco". Catholic Encyclopedia. New York: Robert Appleton Company.
- Chisholm, Hugh, ed. (1911). "Steuco, Agostino". Encyclopædia Britannica. 25 (11th ed.). Cambridge University Press. p. 905.