Since 1901 it has formed part of a national rail link, with the Höllentalbahn and the Ulm–Sigmaringen railway, between Freiburg im Breisgau and Ulm.

The bridges, signal boxes, embankments, gate-keepers; houses, tunnels and stations that were built according to the Baden or Württemberg design "philosophies" are now heritage-listed, but the track infrastructure is not protected.

The connection is particularly known for its attractive location in the Upper Danube Nature Park and on the southern edge of the Swabian Jura, and is considered one of the most scenic railway routes in Germany.

The Tuttlingen–Inzigkofen railway forms part of the shortest rail link between the two cities of Ulm and Freiburg in Baden-Württemberg.

To make matters worse, the Kleber-Express, which had connected Freiburg and Munich since 1954 and had used the entire line from Tuttlingen to Inzigkofen, was discontinued by Deutsche Bahn in 2003.

The situation between the then independent countries of Württemberg and Baden and the Province of Hohenzollern, which had belonged to Prussia since 1850, hampered the construction of the line, which repeatedly crosses the borders of that time.

As in many places, railway committees were founded in the cities and towns along the Danube to promote the construction of such a line.

Since a railway line from Ulm to Vienna already existed at the beginning of the 1860s and Paris was already connected to Chaumont in the east, a line was discussed as the shortest connection between Paris and Vienna that would have closed the gap by running from Ulm along the Danube to Donaueschingen and continuing through the Black Forest to Freiburg im Breisgau, over the Rhine and the Vosges to Chaumont.

A rail connection from the federal fortress of Ulm to the Citadel of Belfort was considered to have central importance.

The existing High Rhine Railway however, passed through the canton of Schaffhausen and Basel and thus through Swiss territory.

The German general staff therefore considered the need for a so-called strategic railways to bypass Swiss territory in the event of war.

From Waldshut to Säckingen trains would then run on the High Rhine Railway, exclusively in German territory.

In 1887, the general staff contractually agreed to close the gap between Inzigkofen and Tuttlingen, and on 26 November 1890, 15 years after the signing of the treaty between Baden and Württemberg, this section was opened for service.

From 1913, express service was also provided from Munich via the Danube Valley Railway to Freiburg, which sometimes featured trains with a dining car.

Freight service was of little significance on the Danube Valley Railway due to the lack of industrialisation along the route.

Towards the end of World War II, Allied air strikes reached the cities and towns along the railway.

In December 1944, Allied bomber squadrons completely destroyed Ulm Hauptbahnhof, as well as the nearby shunting yard at Söflingen.

Heavy damage to the line was then caused by the retreating Wehrmacht, which blew up several railway bridges, making through traffic impossible until 1950, although some sections were operated again from 1946.

In terms of local service, until 1963 it was initially the Württemberg T 5 leading the trains, which started to be replaced from 1961 by the DRG Class 64.

It is served under a two-hourly clock-face timetable, Monday to Friday, at times hourly, with Regional-Express trains running between Ulm and Donaueschingen (some continuing to Villingen).

Between May and October, however, the Naturpark-Express runs on weekends between Gammertingen, Sigmaringen, Tuttlingen and Blumberg-Zollhaus, which provides additional capacity for carrying bicycles.