Middelfart station is located on the main line Copenhagen–Fredericia railway from Copenhagen to Funen and Jutland.

The first station in the town opened in 1865 as the western terminus of the Funen Main Line between Nyborg and Middelfart via Odense.

The first station in the town opened in 1865 as the western terminus of the socalled Queen Louise's Railway, the main line across the island of Funen between Nyborg and Middelfart via Odense.

[4] The station was inaugurated along with the railway line on Queen Louise's 48th birthday the 7 September 1865, with regular traffic commencing the following day.

[6] The following year, on 1 november 1866, a short branch line, the Middelfart–Strib railway line, opened from Middelfart station to the harbour at Strib 5 kilometers north of Middelfart, from where there was connection via railway ferry across the Little Belt to Fredericia in the Jutland peninsula.