In modern organic chemistry the synthesis of chiral enantiopure compounds is an extremely important objective. To achieve this, biocatalysts have emerged as key tools. The broad reactivity of hydrolytic enzymes combined with their ability to discriminate between enantiomers make them ideal catalysts for resolving racemic organic compounds to provide enantiomerically enriched products. This chapter gives an overview of how the enantioselectivity of hydrolases can be exploited to furnish virtually enantiopure compounds via kinetic resolution of racemic compounds and desymmetrisation of symmetric compounds. Enantioselectivity and E-values are discussed. Reversibility of hydrolase-catalysed reactions and how it can be avoided by using various irreversible acyl donors as well as the determination and optimization of enantioselectivity are briefly discussed. The bulk of the chapter consists of selected examples of hydrolase catalysed reactions of some important classes of compounds. Thus the usefulness of hydrolase-catalyzed reactions is demonstrated by several examples of reactions in organic solvents of primary, secondary, tertiary alcohols, various amines, and acid derivatives.