Yeast cells are popular microorganisms for use in various bioprocesses because of their ability to produce various enzymes. They are also known for their low price. However, the activity of these biocatalysts is limited by cell membrane diffusion. One of the possible solutions to this problem is permeabilization, which makes cell walls and membranes of microorganisms more permeable to allow for easier reagent diffusion, while also maintaining properties of the cells, such as their structure and enzymatic activity. The reusability of permeabilized baker’s yeast cells is a major asset that allows them to be successfully used in biotransformation processes. Because of extensive research on this process, numerous procedures have been developed regarding the production of highly active biocatalysts in the form of permeabilized cells. The most commonly used technique is chemical permeabilization. Its effectiveness is based on the activity of various intracellular enzymes: one enzyme or several enzymes simultaneously. Multiple chemical substances, mostly organic solvents and detergents, were analyzed to determine their effectiveness as permeabilizing agents. This review provides a state-of-the-science analysis of substances used as permeabilizing agents for yeast permeabilization