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• Optimization of UAE with single-factor experiments and response surface methodology • Optimal extract obtained with 50% water, 50% amplitude in 17.5 min • Identification of polyphenols by HPLC-ESI-QTOF-MS • Optimal extract exhibited stronger antimicrobial and anticancer activities Actinidia arguta fruit (also known as kiwiberry) has a remarkable phytochemical profile, being reported as a promising preventive agent/treatment for chronic diseases and a desirable source of bioactive molecules for pharmaceutical and cosmetic industries. Despite these promising indications, investigating the full potential of A. arguta fruit remains an unmet need for health applications. This work aims to determine the optimal extraction conditions of antioxidant/antiradical compounds from A. arguta fruit (commonly known as kiwiberry), using the Response Surface Methodology (RSM). The effects of probe amplitude (30%-70%), sonication time (5-30 min) and water:ethanol solvent ratio (0%-100%) were assessed. The optimal extraction conditions were achieved using 50% of water, during 17.5 min and using an amplitude of 50%. A total of 22 compounds, including 6 flavonoids and 4 phenolic acids, were identified in the optimal extract through HPLC-ESI-QTOF-MS, which exhibited outstanding antioxidant and antiradical activities (TPC = 18.705 mg GAE/g dw; FRAP = 186.876 µmol FSE/g dw; ABTS = 16.334 mg AAE/g dw; O 2 ·− IC 50 = 829.384 µg/mL; HOCl IC 50 = 16.895 µg/mL; ROO· = 0.18 µg TE/mg dw). Moreover, it displayed antimicrobial activity against Staphylococcus aureus (MIC = 32 mg/mL) and Pseudomonas gingivalis (MIC = 64 mg/mL) and reduced the growth rate of Escherichia coli. In-vitro assays demonstrated that the optimal extract successfully decreased the viability of two human carcinoma cell lines, namely TR146 and HSC-3, at 500 µg/mL and above 500 µg/mL, respectively. The present study showed that A. arguta fruit is a rich source of compounds with potential to be used for pro-healthy purposes. [Display omitted] [ABSTRACT FROM AUTHOR]