Peroxygenases are attractive biocatalysts for the selective introduction of oxygen into organic molecules under mild conditions with hydrogen peroxide as the oxygen source. In addition to the identification of primary peroxygenases, different classes of enzymes were shown to display promiscuous peroxygenase activity. Even though enzymes with peroxygenase activity are promising industrial biocatalysts, further optimization of their properties is required for their effective use in industrial applications. Here we give a comprehensive overview of enzymes with peroxygenase activity and review diverse strategies, including directed evolution, rational approaches and the assistance of small functional molecules to improve the expression, catalytic activity, substrate scope or selectivity of these promising enzymes. Furthermore, we discuss the exploration of modified or unnatural cofactors to design artificial peroxygenases for desired reactions. The rapidly expanding field of hydrogen peroxide-utilizing enzymes bears a great potential to provide biocatalysts for selective oxyfunctionalization chemistry, contributing to the development of environmentally friendly and sustainable oxidation processes.

过氧合酶是有吸引力的生物催化剂，用于在温和条件下以过氧化氢为氧源将氧选择性地引入有机分子中。除了鉴定初级过氧化酶以外，还显示了不同种类的酶显示混杂的过氧化酶活性。尽管具有过氧合酶活性的酶是有前途的工业生物催化剂，但要使其在工业应用中有效使用，还需要对其性能进行进一步优化。在这里，我们对具有过氧合酶活性的酶进行了全面概述，并综述了各种策略，包括定向进化，合理的方法以及辅助小功能分子以改善这些有前途的酶的表达，催化活性，底物范围或选择性。此外，我们讨论了修饰或非天然辅因子的探索，以设计用于所需反应的人工过氧化酶。利用过氧化氢的酶的迅速发展的领域具有巨大的潜力，可以提供用于选择性氧官能化化学的生物催化剂，有助于发展环境友好和可持续的氧化过程。