Pearl millet, an underutilized crop, is a relatively good source of nutrients and has varied uses over cereals. New applications must be explored to popularize the millet. In the present study, flour and isolated starch from pearl millet cultivars-Kalukombu and Maharashtra Rabi Bajra (MRB)-were investigated for physicochemical properties, nutritionally important starch fractions and x-ray diffraction. The yield of starch was significantly low (Kalukombu: 34.5 g/100 g and MRB: 39.4 g/100 g) with traces of non-starch components (protein, fat, and ash) indicating its purity. Starch could be classified as non-waxy type based on low amylose content (2.86-4.96 g/100 g). Low amylose lead to fragility of swollen starch granules which disintegrated easily at 65°C as observed in swelling power and solubility. Isolated starch was characterized with low water-and oil-holding capacity which could be attributed to the low protein content and absence of fiber in the starch. However, MRB starch showed higher oil uptake compared to flour, possibly due to its larger surface area that increased oil uptake. X-ray diffraction patterns showed sharp peaks at 2θ values 15 and 23º and a diffused peak at 2θ of 17 and 18º which is characteristic of A-type pattern. Low resistant starch (RS) and high readily digestible starch (RDS) content observed in the isolated starch could be attributed to the A-type pattern of starch which is more susceptible to enzyme hydrolysis and also due to elimination of structural obstruction to amylase hydrolysis during the process of starch isolation and gelatinization.