The present research work introduces a novel mixture optimization methodology for biodiesel fuels using an Evolutionary Computation method inspired by biological evolution. Specifically, the optimal biodiesel composition is deduced from the application of a nature-inspired adaptive genetic algorithm that first examines percentages of the ingredients in the optimal mixtures. The innovative approach’s effectiveness lies in problem simulation with improvements in the evaluation of the specific function and the way to define and tune the genetic algorithm. Environmental imperatives in the era of climate change currently impose the optimized production of alternative environmentally friendly biofuels to replace fossil fuels. Biodiesel in particular, appears to be more attractive in recent years, as it originates from renewable bio-derived resources. The main ingredients of the specific biofuel mixture investigated in this research are diesel and biodiesel (100% from bioresources). The assessment of the new biodiesel examined was performed using a fitness function that estimated both the density and cost of the fuel. Beyond the evaluation criterion of cost, density also influences the suitability of this biofuel for commercial use and market sale. The outcomes from the modeling process can be beneficial in saving cost and time for new biodiesel production by using this novel decision-making tool in comparison with randomized laboratory experimentations.