Titanium (Ti) alloys are well-known for their unique combination of properties such as high specific strength at room and elevated temperatures, excellent ductility and good corrosion resistance [1]. Owing to these superior properties, Ti alloys find applications in chemical, biomedical, marine as well as aerospace industries [2]. Amongst the different categories of Ti alloys, a metastable β Ti-5Al-5V-5Mo-3Cr (Ti-5553) alloy is developed for landing gear application in aerospace industries [3]. The present study focuses on assessing the microstructural evolution and generation of mechanical properties of Ti-5553 alloy through cold rolling (CR) technique. Three different degree of deformations i.e. 5%, 10% and 20% thickness reductions are employed on the as-cast (AC) Ti-5553 alloy. The microstructures of Ti-5553-AC, Ti-5553-CR-5, Ti-5553-CR-10 and Ti-5553-CR-20 alloys are analyzed using optical, scanning electron and transmission electron microscopy techniques. Ti-5553-AC alloy exhibits single β phase microstructure, as confirmed using EBSD and X-ray diffraction techniques. On the other hand, cold rolling activates various deformation mechanisms that is related to the extent of deformation employed. While, Ti-5553-CR-5 alloy deforms mainly via slip, Ti-5553-CR-20 alloy shows a combination of parallel slip lines, zigzag slip lines and deformation band. Interestingly, XRD study indicates the occurrence of stress induced martensitic phase for all the cold rolled alloys. EBSD analysis quantifies that volume fraction of this martensite phase is directly related to the degree of deformation. Presence of {332} <113>_β twin along the <110> direction in the β matrix is also observed for the Ti-5553-CR-10 and Ti-5553-CR-20 alloys. These microstructural modification are expected to influence the mechanical properties as well. In this regard, hardness, compressive yield and peak strengths are estimated for the as-cast and different cold rolled alloys. Ti-5553-CR-20 alloy exhibits highest magnitude for these mechanical properties amongst all the conditions. Overall, this study is a unique one in establishing the structure-property correlation for Ti-5553 alloy with respect to the degree of deformation in a systematic way.