Jews settled in Pitigliano in the 15th century. The Jewish population continued to grow as Jews sought refuge there when they were forced out of the Papal State. Protected by the Medici family, the Jewish community flourished and the city became known as "Little Jerusalem" (La Piccola Gerusalemme).

The synagogue was constructed in 1598. A school was built during this period and a plot of land was allotted for a Jewish cemetery.

When Pitigliano became part of the Grand Duchy of Tuscany in 1608, Jewish prosperity and freedom were threatened. Discussions took place on whether the Jews would have to move to the ghettos in Florence or Siena, but, instead, a ghetto was built in Pitigliano.

Similar to other Jews in Tuscany, Jews were forced to wear a specific items of clothing that marked them as a Jew, a red hat for men and a red badge worn on the shirt sleeves for women. Jews, occasionally, had to pay special taxes for various projects, such as the building of a guard house and a fountain.

Despite the restrictions, the Jewish community continued to flourish and prosper. In 1773, Pitigliano had a Jewish population of 200. Jews were involved in all sorts of trades and owned shops selling crafts, cloth and spices.

Napoleon's conquest of Italy in 1799 dramatically changed the situation of the Jews of Pitigliano. Jews felt the repercussions of the Viva Maria riots in Arezzo, when houses in the ghetto were ransacked and fourteen members of the community were arrested.

Soon after this incident, life returned to normal for the Jews of Pitigliano. In 1825, Jews owned 94 houses, 20 warehouses, 11 shops and 10 stables in the town. In 1833, a school and a charity organization were started. Friction arose between the Jewish and Christian communities over the issue of forced conversions of Jewish children. Despite the friction, the Jewish population grew and reached 359 by 1841. At this time, the Jewish community had one rabbi, two vice rabbis and several teachers.

In 1859, the 423 Jews of Pitigliano were emancipated and were granted equal rights. This marked the beginning of the decline of the Jewish population of Pitigliano. Many Jews decided to leave Pitigliano and move to larger cities in Tuscany, while others converted to Christianity.

In 1865, a huge library was built, which housed more than 600 books written in Hebrew.

By 1931, the Jewish population had declined to 70. A massive anti-Semitic campaign began in 1936. Racial laws were instituted in 1938. More Jews left Pitigliano and immigrated, while others were deported.

During the Holocaust, efforts were made to hide the Jews and thwart Nazi effort to kill them. All of the Jews of Pitigliano survived the war.

Today, only three members of the Jewish community are left; nevertheless, the Jewish cultural heritage has been preserved. The synagogue was restored and cultural events take place there; a kosher bakery was also reopened. A kosher version of Pitigliano's famous wine is also being manufactured.