Abstract: Bacterial resistance to penicillin antibiotics can be overcome by making semisynthetic penicillin derived from the 6-amino penicillanic acid (6-APA) raw material. 6-APA is the result of penicillin hydrolysis by the activity of penicillin acylase. Penicillin acylase is an enzyme that catalyzes the hydrolysis of penicillin into 6-amino penicillanic acid (6-APA). The use of penicillin acylase has reached 85% in the pharmaceutical industry in the world. The presence or absence of enzyme penicillin acylase produced by bacteria depends on the type and habitat of bacteria. This research was conducted with several stages, sample isolation, srceening of bacteria, and bacteria identification.