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BACKGROUND Penicillins and cephalosporins are the β-lactam antibiotics most used worldwide for therapeutic treatment of different bacterial infections. Currently, both drugs are produced using penicillin G acylase (PGA) as biocatalyst. OBJECTIVE The objective of this manuscript is to offer an overview of the most recent advances on PGA production and its applications. METHODS A search of scientific reports that include the words " β-lactam antibiotics", "PGA", "PGA production", and "PGA application" was performed. RESULTS PGA producing bacterial strains can be improved through genetic manipulations. However, these genetic manipulations could bring on a potential accumulation of immature precursors involved in the expression of PGA, resulting in low enzyme yield, activity, and/or stability. PGA activity can be enhanced by adding an adequate carbon or nitrogen source to the growth medium at a specific pH or temperature depending on the nature of the bacterial PGA or immobilizing PGA to a solid support to recycle it for a prolonged time Conclusion: In addition to β-lactam antibiotics production, PGA can also be used to obtain key molecules for the synthesis of anti-tumor, antiplatelet, antiemetic, antidepressive, anti-retroviral, antioxidant and antimutagenic drugs.