Duke Nicholas was born at Carlsruhe, Kingdom of Prussia (now Pokój, Poland) was the third child of Duke Eugen of Württemberg (1788–1857, son of Duke Eugen of Württemberg and Princess Louise of Stolberg-Gedern) by his second marriage to Princess Helene of Hohenlohe-Langenburg (1807–1880, daughter of Charles Louis, Prince of Hohenlohe-Langenburg and Countess Amalie of Solms-Baruth).

Nicholas had three half-siblings by his father's previous marriage with Princess Mathilde of Waldeck and Pyrmont.

Since 1888, Duke Nicholas lived in Carlsruhe (now Karlsruhe), where he operated as agriculture and forestry, and carried out studies on the fishing industry.

At his death the third branch of the House of Württemberg (Carlsruhe) became extinct in the male line.

After The King's death Carlsruhe and the headship of the House of Württemberg went to Duke Albrecht (1865–1939).