Zoo spokeswoman Jana Ptacinska Jiratova said the female honey badger named Kaca escaped earlier this week, when she probably climbed over a fence around two meters (six feet) high.
Pavel Brandl, in charge of mammals at the Prague zoo, said the honey badger is considered the most fearless animal in the world.
He said the small, furry creature which weighs about 17 pounds (eight kilograms) and loves honey is unlikely to survive in the wild.
Ptacinska Jiratova says the zoo has received a number of tips from Prague citizens about Kaca's possible whereabouts, but so far the search has been fruitless.
She said the animal, which lives in the wild in Africa, is not dangerous but could become aggressive if she feels threatened.
Prague zoo is one of the few in the world where six honey badgers were born in captivity. The 10-year-old Kaca, who joined her partner Cert in Prague nine years ago, is the mother to all of them. The last of them was born early this year.