Recombinant proteins, obtained through precision fermentation, present promising opportunities as novel ingredients for food applications. However, research into characterising their food-relevant functional properties is limited. The present study investigated the physicochemical properties of recombinant bovine β-casein and compared these properties to those of their native bovine counterpart. Both were characterised in terms of solubility, zeta potential, calcium sensitivity, particle size distribution, radius of gyration, secondary structure, and rheological properties. When dissolved in a 50 mM sodium phosphate buffer, recombinant β-casein had a lower zeta potential over a different range of observed pHs. Its isoelectric point (pI) was shifted to a higher pH of 5.5 compared to 4.5 for native β-casein. Solubility trends generally matched the zeta potential profiles, with the most pronounced difference observed at pH 5.5, where recombinant β-casein showed lower solubility (33.02 ± 0.03 %) compared to native β-casein (78.45 ± 1.22 %). Secondary structures, determined using circular dichroism (CD) and small-angle X-ray scattering (SAXS) measurements, showed similar conformations and radii of gyration for both proteins. Recombinant β-casein demonstrated lower calcium sensitivity and formed stronger self-supporting gels with an earlier onset of acid-induced gelation compared to native β-casein. To further assess its application potential, a cheese model system was developed, demonstrating that recombinant β-casein can form stable, self-supporting gels with desirable texture properties comparable to those of native casein. These results provide the first comprehensive characterisation of the functional properties of recombinant β-casein in food-relevant conditions and highlight its potential use in developing next-generation dairy analogues developed through precision-fermented ingredients.