Globally, the energy demand has risen in the recent years, owing partly to the worlds massive population boom. Fossil fuels, the most often used fuel today, are responsible for a number of environmental issues. Due to this, there is a need in reduction of usage in the fossil fuels, and as a precaution, the researchers paved more attention on the biofuels. For obtaining energy source at a sustainable rate, the biofuels gained attention and it also helps in meeting the demand for energy in the forthcoming days. There are three types of biofuels: 1st generation, 2nd generation and 3rd generation biofuels. Large amount of areas is required for cultivating the food crops and it is termed as first generation or even second generation biofuels. The sources from biofuel which is considered as our third generation biofuel do not act as a reason for any food source and has attracted many researchers. The process of transesterification using microalgae with the addition of various catalysts is widely used for preparing biodiesel. Biodiesel is a renewable fuel that has the possible to replace diesel engines. Due to its biodegradability, oxygenation, sulfur-free nature, and lubricity the best alternate for diesel is explored as biodiesel. This investigation aims at finding out the potential of using bio-oil from chlorella algae. The main focus of the paper is to measure the performance and emission characteristics of B20 Chlorella Algae Methyl Ester (CAME) and to measure the fuel flow and emission characteristics of B20 chlorella algae methyl ester with acetylene injected at 1 L per minute to 4 L per minute at all loads in the increment of 1lpm. © 2025 Elsevier B.V., All rights reserved.