The Turks knew that Belgrade was the greatest obstacle to their conquests in central Europe. After the fall of Smederevo in 1440, Sultan Murad II led an army of over 100,000 Turkish soldiers to besiege Belgrade's fortress. Determined to capture the town, the Turks built their own stronghold on the nearby Žrnov hill, present day Avala, from which they were able to observe and control the surrounding area. Belgrade was able to repel the Turkish attacks for an entire century, until 28 August, 1521. Led by Sultan Suleyman the Magnificent, the Turks finally managed to conquer Belgrade - a key stronghold of Hungary and the Christian world. The town was razed to the ground and burned, thus clearing the path to Western Europe.

With the border now further to the north, Belgrade's strategic significance changed, and the town enjoyed 150 years of relative peace, growing into a major trade and transport hub. The seat of the sancak was relocated from Smederevo to Belgrade and the town began to develop. Crafts and trades flourished in this oriental-looking, newly reconstructed town, a meeting place for traders from Dubrovnik, Venice, Greece and Austria, as well as Turkish, Armenian, Gypsy and Serbian craftsmen. Of the tradesmen, tailors, cooks and armourers were some of the most valued in the town. Near the central town area, numerous caravansaries, covered market places ('bezistans') and mosques were built. The town spread out from beyond its mediaeval ramparts and began to spread along the well-known trade routes, which connected the Orient with Europe. Under the Turks, the town reached a high point in the 17th century, with a population numbering some 100,000.

At the end of the 17th century, Belgrade was struck by plague, which in addition to the fires and revolts by the Janissaries brought the town's growth to a standstill. After 167 years of relatively peaceful development, the town became caught up in the midst of military conflict. Following their September 1688 victory over the Turks in Vienna, the Austrians moved on to conquer Belgrade. Two years later, the Turks retook control of Belgrade, but the town was left in ruins after the conflicts and because of their cooperation with the Austrians, the town's inhabitants were subjected to killings, persecution and pillage. Belgrade yet again became a border town until 1717, when the Austrians, led by Prince Eugene of Savoy, recaptured the town. On the site of the ruined Belgrade fortress, the Austrians built a more modern fortress in response to the strategic military requirements of that time.

The emergence of a Serbian national consciousness and the events surrounding the slaughter of Serbian dukes resulted in Karađorđe leading the First Serbian Uprising of 1804, with the aim of liberating Belgrade. After two years of fighting, Karađorđe and his 25,000 followers liberated the town of Belgrade in 1806 and then the fortress in 1807, both of which they found to have suffered extensive damage. Belgrade became the capital of all the recently liberated areas of Serbia and a symbol of the Serbian people's tradition of freedom. After rebuilding it became an important economic, trade and cultural centre. In 1807, the Serbian government (Praviteljstvujući sovjet) was based in Belgrade and from 1811 the first ministries were established. Prominent people and intellectuals, including Sima Milutinović and Dositej Obradović, who established the first High School (Velika škola) in 1808, came to Belgrade from Vojvodina and other regions.

The Turkish conquest in 1813 interrupted Belgrade's dynamic development, and the repression that followed led to another uprising in 1815. Prince Miloš Obrenović, who led the uprising, managed to introduce more diplomacy into relations with the Turks. By granting special privileges, he enticed more and more Serbs to leave the south and relocate to Belgrade, while the Turks sold their houses and land for far less than they were worth. The Turks kept a hold of the fortress, while the town belonged to the Serbs. Important buildings, such as the residence of Princess Ljubica (Konak kneginje Ljubice), the Cathedral (Saborna crkva) and the palace at Topčider were built during these years. Besides its economic function, Belgrade also became a significant cultural centre. A printing house was relocated from Kragujevac to Belgrade in 1835 and so the newspaper "Novine srpske" began to be published and the Theological School (Bogoslovija) and the first secondary school ('gimnazija' - gymnasium) were opened. The town became a stronghold for the artists of the time such as Vuk Karadžić, Sterija Popović, Joakim Vujić and Dimitrije Davidović.

The presence of the Turkish army in the fortress was an impediment to Belgrade's development. However, during a bombing campaign on the town, the Turks killed a Serbian boy at the infamous Čukur Fountain (Čukur česma), which was the basis for the start of negotiations on the permanent retreat of the Turks from Serbian towns. After 346 years of rule, the Turks permanently retreated from Belgrade on April 18th, 1867. Belgrade was liberated with a symbolic handing-over of the keys, when Serbian soldiers replaced the Turkish guards, as the Serbian flag was raised next to the Turkish flag. Prince Mihailo Obrenović moved the capital from Kragujevac to Belgrade. This provided the fuel for the town's accelerated economic and cultural development.

In the second half of the 19th century, city planning gave Belgrade a more European appearance. Knez Mihaila street (ulica Kneza Mihaila) became the central focus of the city, as it was the shortest connection between the fortress and the town, and quickly became the main trade and business area, which it remains today. Traditional crafts were replaced by industry and the construction of the Belgrade-Niš railway line in 1884 improved trade and transport. Electricity was introduced to the city, alongside trams and a steamboat society. Important scientific and cultural institutions were also established in Belgrade, such as the High School (Velika škola) in 1863 and the National Theatre (Narodno pozorište) in 1869.