The diversification of the energy matrix, combined with the use of renewable and less polluting fuels in internal combustion engines, has encouraged numerous research efforts both nationally and internationally. In this context, the utilization of waste for biofuel production stands out as a promising alternative, offering a clean and economically viable energy source. Biogas is one of the most sustainable options and has been widely used in the industry. However, it presents low lower heating values (LHV) and difficulties in burning stoichiometric mixtures, which compromise engine performance, resulting in higher specific fuel consumption and lower power output compared to fossil fuels. To address this challenge, this study aimed to improve biogas combustion in internal combustion engines by investigating the application of a new pre-chamber ignition system in the combustion process and engine performance parameters. For this, experimental tests were conducted with two biofuel concentrations for evaluation: (100% CNG) and (85% CNG + 15% CO2), enriching the stoichiometric mixture and applying calibration methodologies in a single-cylinder engine adapted to operate with biogas, assessing engine performance parameters and gas emissions. The application of pre-chamber ignition showed significant improvements in energy efficiency, resulting in approximately a 12% torque gain in stoichiometric mixtures, contributing to more efficient combustion and a reduction in hydrocarbon emissions. The use of mixtures in the range of 1.0 to 1.2 led to emission reductions between 60% and 35% compared to the engine without the pre-chamber, demonstrating the pre-chamber’s ability to promote more complete combustion even in leaner mixtures. The data obtained provide valuable insights for the development and application of new technologies in biogas-powered internal combustion engines, contributing to advancements in this research area.