Asphalt pavements are the most popular type of pavements used in roads. Nowadays, asphalt pavement is used in road construction more than 90 percent. Bituminous materials obtained from oil sources are the main determinant factor of the quality and cost for building asphalt pavement. Due to the limited resources of oil and price increase in crude oil and bitumen, the construction cost of asphalt pavements has been increased, so using alternative materials instead of bitumen is logical. In recent years, reusing waste materials produced in industrial processes as the primary raw material in other manufacturing processes has became one of the major environmental goals in scientific and technological research. Vinasse biopolymer is considered as a waste industrial material during ethanol production from sugarcane. The huge volume of vinasse waste has turned it into one of the main concerns. Vinasse is a natural biopolymer, biocompatible which has a cohesive property. Due to the low cost of vinasse compared to bitumen and its similarity in terms of chemical structure to the bitumen, it can be considered as an alternative for bitumen. Mixing of vinasse with bitumen is carried out easily since it is liquid in ambient temperature. Using natural and renewable vinasse in bitumen is an efficient and sustainable way in order to reducing costs for constructing asphalt pavement and positive step towards protecting the environment by disposing this waste. In this study, physical and mechanical properties of bitumen have been investigated by addition of vinasse biopolymer to bitumen. Basic tests such as penetration degree, softening point, ductility, flash point, kinematic viscosity at 135°C and weight loss (oven 163°C) were conducted on bitumen samples containing vinasse (0, 2, 5 and 8 wt.%). The results of basic tests showed an increase in penetration degree, kinematic viscosity and weight loss in sample reinforced 8% vinasse compared to the neat bitumen, and a decrease in ductility and flash point of all reinforced samples compared to neat bitumen, and at the same time the required quantities of standards were kept safetly. Also, the temperature sensitivity of bitumen was decreased by addition of 8% vinasse. Performance tests (SHRP) such as dynamic shear rheometer (DSR), elastic recovery and multiple stress creep recovery (MSCR) were conducted on base bitumen and samples containing 5% and 8% vinasse biopolymer. According to the DSR test results the ability of bitumen to maintain fatigue and rutting resistance, as well as performance grade of reinforced bitumen has not changed and high operating temperature for neat bitumen and samples containing biopolymer has been estimated between 63°C to 65°C. The results of MSCR test indicate the possibility of using high traffic levels, reducing the sensitivity to applied stress compared to the neat bitumen, as well as maintaining the required values of standards based on MSCR test in samples containing 5% and 8% vinasse biopolymer. The addition of biopolymer has increased the rate of elastic recovery by more than four times and increased bituminous elastic domain. Bituminous chemical structure was investigated by performing Fourier transform infrared spectroscopy (FTIR) experiments on bitumen and bitumen samples containing 8% vinasse. The results of this experiment show that adding 8% of biopolymer to bitumen did not change chemical structure of bitumen but reduces oxidation indices. Alcohol contained in biopolymer also has no effect on chemical structure of bitumen. Investigation of samples containing 0, 2, 5 and 8% of vinasse with polarizing microscopy with a magnification of 100 degree indicated homogeneous and uniform mixing of biopolymer with bitumen. Key Words bitumen, vinasse biopolymer, alternative additive, rheological properties of bitumen, physical and mechanical properties of bitumen.