Nanocomposite consisting of 5 mol% crystalline CuI-doped hydroxyapatite (HA) was prepared for the first time by facile chemical method and calcined at different temperatures such as 300 °C, 500 °C, 700 °C and 900 °C. In this study, HA played a role as the matrix and CuI was the reinforcement. The effect of calcination temperature on XRD pattern, TEM, SEM, BET, FTIR, luminescence intensity, photoluminescence quantum yield (PLQY), fluorescence lifetime, charge mobility measurements, electronic speckle pattern interferometry and mechanical properties were investigated. The modified Scherrer equation of the composites showed that the size of the particles increased with increasing calcination temperature from 66.64 to 87.20 nm. The photoluminescence intensity of CuI (5 mol%)/HA was quenched at room temperature, while the represented constant decay time for the calcined compound was increased at 700 °C. Furthermore, the CIE coordinates were shown 0.39333, 0.18493 for CuI (5 mol%)/HA calcined at 700 °C. The charge mobility values of the nanocomposites were extracted by space-charge limited current (SCLC) method and the range of effective mobility was from 3.645 × 10−4 to 6.697 × 10−4 cm2V−1s−1. Mechanical properties were fully discussed via ASTM-E9 standard and CuI (5 mol%)/HA calcined at 900 °C was in better range than other compounds, the σyc and hardness values were reported 7.32 Mpa and 40.81 HV respectively. Speckle interferometry was used to demonstrate that there was no large imperfection in the surface of each sample. The components were very sensitive to the calcination temperatures. In other words, the combination of CuI (5 mol%)/HA showed that the replacement of Cu ions with Ca was helpful to improve the photophysical and mechanical properties.