John V of Anhalt-Zerbst (Dessau, 4 September 1504 – Zerbst, 4 February 1551), is a German prince of the House of Ascania and ruler of the principality of Anhalt-Dessau.

From 1544, he assumes rule of the re-created principality of Anhalt-Zerbst.

John is the second (but eldest surviving) son of Ernest I, Prince of Anhalt-Dessau, by his wife Margarete, daughter of Henry I, Duke of Münsterberg-Oels, and granddaughter of George of Poděbrady, King of Bohemia.