In 1785, merchants and townspeople of the Merchant Suburb of Kherson appealed to Archbishop Nikifor with a request to build a church in honor of the Assumption of the Blessed Virgin Mary with two chapels: Saint Nicholas the Wonderworker and Saint Barbara. This was necessary as the existing Church of Saint Sophia was Greek, and services were conducted in Greek. The church was laid in 1786 in the center of the suburb on Greek Square. The main benefactors were merchants Pyotr Fendrikov and Ivan Sizarev. The Russo-Turkish War of 1787 delayed construction, so a wooden church of Saint Archangel Michael was temporarily moved, consecrated in 1788. Ten years later, it was dismantled, as on April 14, 1798, the Assumption Church was consecrated. It was cross-domed, 46 meters long, 31 meters wide, and 40.5 meters high. A wooden bell tower 45 meters high was added. The chapels were consecrated later: the southern in honor of Nicholas the Wonderworker on May 16, 1799, and the northern in honor of Saint Barbara. According to legend, the construction was completed thanks to a donation from one of the Zaporozhians. In 1828, after the transfer of the Kherson Admiralty institutions to Mykolaiv, the Assumption Church became the city cathedral. In 1833, the wooden bell tower was replaced with a stone one, and in 1897, an iron spire with a cross was installed on it.

Over time, an ensemble of church buildings grew around the temple. In front of the entrance, from the side of Rishelievskaya Street, two two-story houses were built: in 1822 to the north and in 1829 to the south. The first was rented out, while the second housed the clergy and the Spiritual Administration, and from 1849 to 1874, the Spiritual School.

In 1853, Archbishop Innocent achieved the opening of a vicarage in the Kherson province, and the Assumption Cathedral became a cathedral. The first vicar bishop was Polycarp, who held this position from July 5, 1853, to July 12, 1858. In 1859-1861, the cathedral underwent major repairs, and a fence with a metal grille and three gates appeared around it.

In 1867-1868, a two-story house for priests was built, and in 1896, a reading-library house and an auditorium for religious-moral readings. In the 1850s, two important events occurred: on May 8, 1852, the first procession with the Kasperov Icon of the Mother of God arrived, and on October 26, 1855, Emperor Alexander II visited the cathedral. After the end of the Crimean War, he sent a letter of gratitude to the residents of the Kherson province. Among the benefactors of the cathedral were known figures such as Yakov Doroshenko, Ivan Volokhin and his son Alexander Ivanovich Volokhin, as well as merchant Daniil Galukhin.

The Assumption Cathedral, the largest in Kherson, was simple in architecture but richly decorated inside. The grand iconostasis, 17 meters high, contained 50 icons. Only one, the Blessed Virgin Mary, has survived. The first icons were likely painted by Kyiv masters. In the late 1850s, Timofey Tolmachevsky painted the interior in Byzantine style, and Kherson artist Z.I. Ryabkov painted the chapels and frescoes of the church. None of this has survived.

In the Soviet years, the cathedral faced the fate of many churches. In 1922, valuables were confiscated, in 1929 the bells were removed, and in 1930 the cathedral was closed and repurposed as a sports club, destroying the interior. During the occupation, the church operated, but in 1944 it was closed again and turned into a Palace of Sports. From 1945 to 1993, the cathedral housed the sports club 'Petrovets'. In September 1993, the church was returned to the Ukrainian Orthodox Church and restoration began.

The Assumption Cathedral is a monument of 18th-century architecture and is part of the protected zone of the city's historical development.