Biodiesel is an alternative, sustainable, eco-friendly, clean, biodegradable fuel that helps to get rid of fossil fuels that are not only depleting but also causing health issues for humans and environment. In present study, waste animal fats (fish and chicken waste) and aquatic weeds (Eichhornia crassipes, Pistia stratiotes and Lemna minor) were utilized as oil source for biodiesel production. Highest lipid content shown by fish waste oil that was 36% then followed by 33% in chicken waste oil, 16.7% in Eichhornia crassipies oil, 12.6% in Lemna minor oil and 4.11% in Pistia Stratiotes oil. Fatty acids converted into fatty acid methyl esters (biodiesel) through transesterification method. Then characterized by GCMS that confirmed the presence of highest percentages like 51% of oleic acid in Chicken waste oil biodiesel, 35.7% of palmitic acid in Fish waste oil biodiesel, 14.6% of palmitoleic acid in Eichhornia crassipies oil biodiesel, 61.7% of phthalic acid in Lemna minor oil biodiesel and 10.9% of linoleic acid in Pistia Stratiotes oil biodiesel. Physicochemical analysis of each biodiesel confirmed within the ASTM standard. Emission performance of biodiesel and petro-diesel blends showed maximum decrement in CO (26%, 22%, 24% and 23%), NO (90%, 80%, 60% and 60%), SO2 (70%, 87%, 87% and 83%)and smoke (25%, 50%, 25% and 62%) were observed in fish waste oil biodiesel, Eichhornia crassipies oil biodiesel, Pistia Stratiotes oil biodiesel and Lemna minor oil biodiesel respectively. Similarly increment in CO2 (3.75%, 3.8%, 3.72%, and 3.8%) and O2 (11.62%, 11.32%, 11.57% and 11.68%) was observed respectively because biodiesel have more oxygen content that make the complete combustion of fuel. Thus, biodiesel can adopted as an alternative fuel having less environment impact.