The currently known methods of enzymatic beta-lactam synthesis, as well as the enzymes and heterogeneous biocatalysts used for this purpose, are presented, and the published reports on advances in the field of enzymatic synthesis of selected antibiotics belonging to the groups of penicillin acids and cephalosporin acids are summarized in the present review. The key conditions and parameters of biocatalytic processes, such as the biocatalyst form, concentration of the precursor compounds, solvent type, pH, temperature, etc. are analyzed and compared, and guidelines for further optimization of beta-lactam synthesis are given. The present review may be of use for a wide range of readers, as well as to enzymology and biotechnology experts.