Mineralized collagen provides new solutions in the manufacture of porous scaffolds for tissue engineering. In this research, the use of biomimetic mineralization, the process of mineralization of acid soluble collagen (ASC) type I extracted from white fish (Rutilus frisii kutum) scales, in a body simulated fluid (SBF) in the presence and absence of an anionic surfactant, sodium dodecyl benzene sulphonate (SDBS) was thoroughly investigated. The formation of β-tricalcium phosphate (β-TCP) nanocrystals in mineralized nanocomposites was detected by XRD, TGA, and FT-IR techniques. The denaturation temperature (Td) of mineralized ASC was determined at 42.87 °C without surfactant, and at 70.2 °C with surfactant using DSC technique, which indicated a higher thermal stability of ASC without the SDBS surfactant. TEM images illustrated that the formation of β-TCP crystals on collagen nano-fibrils was in an irregular form in the absence of surfactant while it was more regular and in the form of rectangular sheets in the presence of surfactant. The results of UV/Vis spectrophotometer for turbidity measurements during the initial step of biomineralization revealed that SDBS while accelerating the self-assembly process of collagen nano-fibrills, delayed the process of nucleation and the growth of calcium phosphate nanocrystals by increasing the kinetic energy barrier.