Police, who believe the animal is an escaped pet, were first alerted around midnight by two members of the public who recorded mobile phone footage of what appeared to be a wild boar and a lion chasing each other. No boar carcass has been found.
Authorities believe the wild cat could currently be asleep in one of the many lakeland forests in Brandenburg, the state that surrounds Germany's capital.
An operation to track it down using two helicopters, drones and infrared cameras was expanded in the early hours of Thursday as a hundred police officers joined forces with hunters and vets.
They would aim to tranquilise and capture the animal and only kill her if she posed a danger to people, Gruber said, adding that the search was now concentrated in the municipality's northeastern part.
Since no zoos or circuses have reported a missing lioness, police believe she must be an escaped pet.
Animal rights groups criticised successive governments for failing to ban the practice of keeping wild animals as pets.
Circus-keeper Michel Ramon Rogall told Reuters there was no circus with wild animals currently on the road in eastern Germany, "and they wouldn't escape either (if there was)."