Experimental and epidemiological studies show a positive relation between consumption of citrus juices and reduction of risk for some chronic disorders, such as diabetes and cardiovascular diseases. In particular, the bergamot juice is characterized by noticeable amounts of phytochemicals such as flavanone glycosides, limonoids, and quaternary ammonium compounds, all health-beneficial biomolecules. In vitro and in vivo studies have shown anti-inflammatory, cholesterol-lowering, and anti-diabetic activities attributed to these compounds depending on their chemical structure. However, nutritional content of bergamot juice may vary as consequence of different processing techniques, thus needing to address this claim. For this reason, the objective of this research was to evaluate the effects of different processing systems on the proximate constituents, the composition, and the antioxidant activity of the correspondent juices. Overall, the results indicate that the process employed may influence the chemical composition and the functional properties of the ended juice. Screw press method produced a juice with greater content of flavanone glycosides (ranged from 37 to 402 mg/L) and limonoid aglycones (ranged from 65 to 67 mg/L) than the other processes (p < 0.001). However, the process used for extraction of bergamot juice did not affect significantly the N,N-dimethyl-L-proline content (p < 0.5). Moreover, the screw press juice showed the highest antioxidant activity with EC50 value of 9.35 µg/mL, thus suggesting that this method maintains for health the nutritional quality of a fresh-pressed juice.