Otto I was Count of Burgundy from 1190 to his death and briefly Count of Luxembourg from 1196 to 1197. He was the fourth son of Frederick I, Holy Roman Emperor by his second wife Beatrice I, Countess of Burgundy, daughter of Count Renaud III.
Upon the death of his mother in 1184, his father granted him the Burgundian county, elevating him to the rank of a Count palatine (Freigraf). Haughty Otto however soon entered into several feuds: not only with the Anscarid lords of Auxonne and Mâcon, who claimed late Beatrice's heritage, but also with the Counts of Montbéliard, the French Odo III, Duke of Burgundy and Berthold V, Duke of Zähringen. In the course of negotiations in 1195, he killed Count Amadeus of Montbèliard with his own hands, followed by the assassination of Alsatian Count Ulric of Ferrette in 1197 and the execution of a brother of Konrad von Hüneburg, Bishop of Strasbourg, in 1198.
When Henry IV, Count of Luxembourg died without heirs in 1196, his county escheated to the Emperor and Henry VI enfeoffed his brother Otto. Theobald I, Count of Bar, who had married Ermesinde, daughter of late Count Henry IV, negotiated the renunciation of Luxembourg with Otto the next year.
Meanwhile, Count Palatine Otto's regional conflicts had become a severe threat to the power politics of his Hohenstaufen relatives. After Philip of Swabia had been elected King of the Romans in 1198, rivaling with the Welf duke Otto of Brunswick, he tried to settle the numerous quarrels picked by his brother. When i 1200 Otto was assassinated at Besançon, his death came in useful to many political actors.
Otto was buried at St Stephen's Cathedral, today the site of the Citadel of Besançon.