The research and development of advanced biofuels is motivated by the reduction of negative environmental impact caused with the use of fossil fuels throughout the centuries, and encouraged with the legal regulations of European Union, that promote energy diversification, decarbonization of the traffic sector, as well as the principles of circular economy. One of such biofuels is biodiesel, that is produced via alcohol transesterification of a certain feedstock, in the presence of a catalyst. Post purification, it is possible to blend it with fossil fuel, if it fulfills the requirements set by the diesel (HRN EN 590) and biodiesel (HRN EN 14214) standard, and use as a renewable source of energy in diesel engines. In the Republic of Croatia it is allowed to blend up to 7 vol% of biodiesel with diesel fuel for their blend to still be declared as diesel fuel. It is possible to blend higher volume fractions of biodiesel fuel with diesel fuel, as well, however these are referred to as the diesel-biodiesel blends. Since the commercial European biodiesel is most commonly produced from partly renewable methanol, the goal of this doctoral thesis was to investigate the influence of the reactant type and the reaction conditions on the biodiesel synthesis from isomers of selected higher alcohols (from propanol to octanol), which can, as bioalcohols, be completely produced via fermentation from biomass. In the first part of this doctoral thesis, the influence of reaction conditions (temperature, reaction time, molar ratio of the reactants, and mass fraction of the catalyst) on the reaction conversion of the biodiesel systems, was studied. With the use of Design of Experiments by Box-Behnken and Response Surface Methodology, the biodiesel synthesis from either sunflower oil and alcohols 1-propanol or 1-octanol, or waste cooking oil and 2-propanol, 2-butanol, 2-pentanol, or 2-octanol, was analyzed and optimized, in the presence of a catalyst potassium hydroxide. Additionally, the dependence of the reaction conversion on the mentioned reaction conditions, the chemical structure, and the molar mass of the reactants, in the presence of potassium hydroxide, was studied, with the selected feedstock being sunflower oil, rapeseed oil, coconut oil, waste cooking oil, or animal fat, that reacted with 1-propanol, 1-butanol, isobutanol, 1-pentanol, isopentanol, 1-hexanol, or 2-hexanol. In the second part of the thesis, larger amounts of biodiesels (fatty acid methyl, propyl, butyl, isobutyl, pentyl, hexyl, and octyl esters) were synthesized from waste cooking oil, in the presence of KOH, at the conditions of the highest experimental conversions; purified, and blended into the binary and trinary blends with non-additivated mineral diesel and/or corresponding alcohol. The prepared samples underwent the application property analysis via corresponding standardized methods (density, kinematic viscosity, cloud point, cold filter plugging point, pour point, lubricity, simulated distillation), the results of which were compared with the ones prescribed by the standard for diesel fuel HRN EN 590. Additionallly, samples were analyzed via instrumental analytical methods: differential scanning calorimetry, surface tension measurements via goniometer, and friction coefficient measurements. The main results showed that the most significant reaction conditions in the given systems were the molar ratio of the reactants, and the mass fraction of the catalyst KOH, and that in the selected systems experimentally the highest conversions were obtained using coconut oil, while the lowest using animal fat, as a feedstock. The reaction conversions in the synthesis using straight-chain alcohols were generally higher than those with branched alcohols, and the increase in the molecular weight of branched alcohols further led to a decrease in the conversion. Sample characterization determined the addition of selected biodiesel fuels up to 20 vol% into the blends with mineral diesel and/or corresponding alcohol led to the increase in their density, kinematic viscosity, and surface tension, while the addition of the selected higher alcohol up to 15 vol% in the blends with diesel and biodiesel resulted in the reduction of their density, kinematic viscosity, and surface tension. Density and kinematic viscosity values of all the binary diesel-biodiesel (with up to 20 vol% of biodiesel), and trinary diesel-biodiesel-corresponding alcohol blends (with up to 15 vol% of biodiesel) were within the limits required by the standard for diesel fuel HRN EN 590. Furthermore, blending of selected biodiesels and/or corresponding alcohols with mineral diesel did not aggravate its analyzed low-temperature properties (cloud point, cold filter plugging point, pour point). Rheological measurements determined that the addition of fatty acid hexyl esters to non-additivated mineral diesel of just 2.5 vol% significantly improved its lubricity, whereas the change in the volume fraction or the type of biodiesel blended with mineral diesel did not have a significant influence on the diesel’s lubricating properties. Finally, it was observed that for the distillation of the same percentage of the sample, the higher temperatures were required with the addition of fatty acid pentyl esters of 20 vol% to mineral diesel, than with the addition of the same biodiesel in the volume fraction of 10 vol%. On the other hand, differential scanning calorimetry showed that the binary and trinary blends of all biodiesels with mineral diesel and/or corresponding alcohol had onset and peak crystallization temperature values close to the ones of non-additivated mineral diesel. This could serve as an additional indicator for the lack of their negative impact on the diesel’s low-temperature properties.