In 1804, Metropolitan Gavriil, while in Odessa, met Moldovan nobleman Alexander Teutul, whose estate, consisting of two plots of 25 desyatins, was located on the seashore in the area of Bolshoy Fontan. Alexander Teutul repeatedly expressed to the metropolitan his desire to build a church and a lighthouse there. In 1814, he handed Metropolitan Gavriil a written certificate of the transfer of his Fontan estate to the Chisinau Metropolis. The history of this generous donation is full of tragedy. One evening, Teutul lit bonfires on the high seashore. A Greek ship heading to Odessa mistook the lights for a lighthouse, went off course, and crashed, resulting in the death of several people. Shocked by what happened, Teutul, seeing a connection with his unfulfilled vow, gifted the surviving sailors and donated his estate to the Church for the construction of a temple and a lighthouse to prevent similar tragedies. Metropolitan Gavriil appealed to the Odessa governor Duke de Richelieu, and soon permission was obtained to formalize the deed for the donated land.

By 1837, the monastery owned more than 158 desyatins of land, which amounted to 173 hectares. In the second half of the 19th century, its land holdings increased, and by 1910 they reached 356 desyatins, almost 390 hectares. Currently, the monastery owns just over 6 hectares of land. During the Crimean War of 1853-1856, the brotherhood of the Holy Uspensky Odessa Monastery prayed for protection from the 'invasion of foreigners,' for peace, the preservation of the Orthodox faith, and the salvation of souls.

In the 19th century, Odessa became a major city, partly because it was a route for pilgrims to the Orthodox East: to the Holy Land and Mount Athos. Odessa was called the 'gates of Jerusalem,' and it housed the dependencies of Athos monasteries: Panteleimon, Andreevsky, and Ilyinsky. Because of this, the Holy Uspensky Odessa Monastery held significant importance for Orthodox pilgrims, for the life of the city, and for all of southern Russia, having a beneficial influence on the spiritual and cultural development of the Black Sea region.

Monastic life in the Holy Uspensky Odessa Monastery continued until 1922. After the monastery was closed in 1922, the inhabitants were dispersed throughout Russia. In 1944, its revival began. In 1946, the monastery was visited by His Holiness Patriarch Alexy I of Moscow and all Russia, and from that time, a summer residence for the Patriarch was established here, who began to visit the monastery annually. Due to this, the monastery began to be called Patriarchal.

In 1988, the year of the 1000th anniversary of the Baptism of Rus, a helmet-shaped dome was installed over the Holy Nicholas Church, and the bell tower was rebuilt.

The monastery reached its true flourishing under Metropolitan Agafangel, who actively worked on beautifying the monastery. Under his leadership, a new cathedral in honor of the icon of the Mother of God 'Life-Giving Spring' and a bell tower with a church in honor of Saints Boris and Gleb were built, becoming a masterpiece of Odessa architecture.

In 1964, the former Athos monk, renowned elder schema-hieromonk Kuksha Velichko, known for his feats of faith and gift of foresight, passed away in the monastery.

In 2006, a wooden chapel in honor of the holy martyrs Faith, Hope, Love, and their mother Sophia was built in the monastery cemetery, where the unceasing Psalter is now read for the living and the dead.