Microfluidization is a unique high-pressure homogenization technique combining various forces such as high-velocity impact, high-frequency vibration, instantaneous pressure drop, intense shear rate, and hydrodynamic cavitation. Even though it is mainly used on emulsion-based systems and known for its effects on particle size and surface area, it also significantly alters physicochemical and functional properties of macromolecules including hydration properties, solubility, viscosity, cation-exchange capacity, rheological properties, and bioavailability. Besides, the transformation of structure and conformation due to the combined effects of microfluidization modifies the material characteristics that can be a base for new innovative food formulations. Therefore, microfluidization is being commonly used in the food industry for various purposes including the formation of micro- and nano-sized emulsions, encapsulation of easily degradable bioactive compounds, and improvement in functional properties of proteins, polysaccharides, and dietary fibers. Although the extent of modification through microfluidization depends on processing conditions (e.g., pressure, number of passes, solvent), the nature of the material to be processed also changes the outcomes significantly. Therefore, it is important to understand the effects of microfluidization on each food component. Overall, this review paper provides an overview of microfluidization treatment, summarizes the applications on macromolecules with specific examples, and presents the existing problems.