This study evaluates the potential of Ultra High-Pressure Homogenization (UHPH) to enhance fruit juices' safety, quality, and sensory attributes, presenting a promising alternative to traditional thermal pasteurization. The effects of UHPH on E. coli K12 inactivation were investigated across four types of fruit juices —watermelon, cantaloupe, blueberry, and grapefruit—assessing the microbial safety immediately post-treatment. Additionally, the study analyzed the impact of UHPH on key physicochemical properties (pH, titratable acidity, Brix, viscosity, turbidity, color, and particle size distribution) and the natural microflora of each juice immediately post-treatment and over a 45-day refrigerated storage period. These effects were compared with those observed under conventional thermal pasteurization and untreated control to evaluate the stability and quality retention of UHPH-treated juices over time. Finally, consumer acceptance of UHPH-treated juices was compared with thermally pasteurized and untreated fresh juices to determine sensory preferences and potential market acceptance.The results demonstrate that UHPH achieved a 5-log CFU/mL reduction of E. coli K12 across all juice types, with higher pressures (250–300 MPa) and higher inlet temperatures (22°C) reducing E. coli to undetectable levels. UHPH-treated juices also showed better stability in physicochemical properties, such as pH, acidity, and turbidity, over the 45-day storage period, with minimal loss in freshness and sensory qualities. Treatment combinations involving higher pressures (250–300 MPa), higher inlet temperatures (22°C), and intermediate flow rates (1.125 L/min) were particularly effective in preserving juice quality, maintaining color vibrancy, and reducing sedimentation. Consumer acceptance testing further highlighted a clear preference for UHPH-treated juices over thermally pasteurized ones, especially for appearance, flavor, mouthfeel and overall acceptance, suggesting that UHPH better preserves the fresh-like and natural-like characteristics of juices. These findings support the adoption of UHPH as an alternative processing technique that meets microbial safety standards and enhances physicochemical stability and consumer acceptability in fruit juices, making it a viable alternative to conventional pasteurization methods.