Isoprenoids are a large class of natural products with myriad applications as bioactive and commercial compounds. Their diverse structures are derived from the biosynthetic assembly and tailoring of their scaffolds, ultimately constructed from two C5 hemiterpene building blocks. The modular logic of these platforms can be harnessed to improve titers of valuable isoprenoids in diverse hosts and to produce new-to-nature compounds. Often, this process is facilitated by the substrate or product promiscuity of the component enzymes, which can be leveraged to produce novel isoprenoids. To complement rational enhancements and even re-programming of isoprenoid biosynthesis, high-throughput approaches that rely on searching through large enzymatic libraries are being developed. This review summarizes recent advances and strategies related to isoprenoid synthetic biology, combinatorial biosynthesis, and chemo-enzymatic synthesis, focusing on the past 5 years. Emerging applications of cell-free biosynthesis and high-throughput tools are included that culminate in a discussion of the future outlook and perspective of isoprenoid biosynthetic engineering.

类异戊二烯是一大类天然产物，作为生物活性和商业化合物具有多种应用。它们的多样化结构源自生物合成组装和支架的剪裁，最终由两个 C5 半萜结构单元构建。这些平台的模块化逻辑可用于提高不同宿主中有价值的类异戊二烯的滴度，并生产新的天然化合物。通常，该过程是由组分酶的底物或产物混杂促进的，可以利用这些酶来生产新型类异戊二烯。为了补充类异戊二烯生物合成的合理增强甚至重新编程，正在开发依赖于大型酶库搜索的高通量方法。本文综述了类异戊二烯合成生物学、组合生物合成和化学酶合成相关的最新进展和策略，重点关注过去 5 年。其中包括无细胞生物合成和高通量工具的新兴应用，最终讨论了类异戊二烯生物合成工程的未来前景和前景。