In 1859, the Orenburg City Administration provided the women's community with 10 desyatins of land for free, north of the Christian cemetery. On this land, merchants Nikolai and Stefan Mikhailovich Deev built several buildings. By 1864, the number of sisters had increased so much that the construction of the first capital building, the "white" cell building, began, completed in 1865. On February 17, 1866, the Assumption Women's Community was officially opened, and in honor of this event, the construction of the home church of St. Nicholas began, consecrated in 1867. On October 20, 1867, the community received permission for its own seal. On August 9, 1867, the abbess of the community, nun Tatiana Alexeyevna Konnova, was tonsured under the name of Thais. By decree of the Holy Synod on September 28, 1872, the community was renamed the Assumption Monastery. According to other sources, this occurred on January 15, 1874. On February 24, 1874, nun Thais was elevated to the rank of abbess and given the staff.

In 1868, the construction of the main two-story church of the Assumption of the Blessed Virgin Mary began, designed by architect K.A. Ton. The church was built on charitable donations and funds collected by Orenburg nuns throughout Russia. The first altar was consecrated on September 14, 1875, and the last one on September 30, 1884. In 1885, the construction of a five-tiered bell tower began, completed in 1888. On its second floor, a church in honor of the Iveron Icon of the Mother of God was arranged in 1893, and on September 12, 1906, tower clocks were installed on the bell tower thanks to the funds and efforts of hieromonk Vitaly. The first abbess Thais, who passed away in 1891, is considered the founder of the monastery, while her successor, abbess Innocentia, is regarded as the builder. Her work was recognized with awards, including the Golden Pectoral Cross with decorations, awarded on May 6, 1912.

After the revolution, in the 1920s, the monastery buildings were used by GUBON and FACT to house orphanages, a medical college, workshops, a school, a children's town, and a party school. From November 1926, all buildings were leased to the Military School of Air Combat for housing pilots' families. The church buildings and stone fence were dismantled, family crypts and graves in the cemetery were destroyed, where military vehicles stood for a long time. Only a few bushes remained from the lilac gardens. Stone production and residential buildings were constructed on the territory. In 1998, due to the construction of a multi-story residential building, the buildings of the livestock yard and stone wing were demolished.

In the post-war Soviet period, the monastery territory was managed by the Ministry of Defense of the Russian Federation. Only in 2010, after long appeals from the Russian Orthodox Church, was the building of the monastery refectory returned. By the blessing of Metropolitan Valentin, nun Flora had been working since December 2006 on the documentation and negotiations with "Rosimushchestvo" and the Ministry of Defense for the return of the buildings and land belonging to the Holy Assumption Orenburg Women's Monastery. In August 2010, the refectory building was returned.