Acies Bio d.o.o., Tehnološki Park 21, SI-1000 Ljubljana, Slovenia.
Metab Eng. 2012 Jan;14(1):39-46. doi: 10.1016/j.ymben.2011.11.003. Epub 2011 Nov 12.
FK506, a widely used immunosuppressant, is produced by industrial fermentation processes using various Streptomyces species. Independently of the strain, structurally related compound FK520 is co-produced, resulting in complex and costly isolation procedures. In this paper, we report a chemobiosynthetic approach for exclusive biosynthesis of FK506. This approach is based on the Streptomyces tsukubaensis strain with inactivated allR gene, a homologue of crotonyl-CoA carboxylase/reductase, encoded in the FK506 biosynthetic cluster. This strain produces neither FK506 nor FK520; however, if allylmalonyl-S-N-acetylcysteamine precursor is added to cultivation broth, the production of FK506 is reestablished without FK506-related by-products. Using a combination of metabolic engineering and chemobiosynthetic approach, we achieved exclusive production of FK506, representing a significant step towards development of an advanced industrial bioprocess.
FK506 是一种广泛使用的免疫抑制剂，通过使用各种链霉菌属（Streptomyces）物种的工业发酵工艺生产。无论菌株如何，都会共同产生结构相关的化合物 FK520，导致复杂且昂贵的分离程序。在本文中，我们报告了一种用于 FK506 专属性生物合成的化学生物合成方法。该方法基于链霉菌属（Streptomyces）筑波变种（Streptomyces tsukubaensis），其 allR 基因失活，该基因是 FK506 生物合成簇中编码的丙二酰辅酶 A 羧化酶/还原酶的同源物。该菌株既不产生 FK506 也不产生 FK520；然而，如果将烯丙基丙二酰-S-N-乙酰半胱氨酸前体添加到培养物中，则会重新建立 FK506 的产生，而没有 FK506 相关的副产物。通过代谢工程和化学生物合成方法的结合，我们实现了 FK506 的专属性生产，这是朝着开发先进的工业生物工艺迈出的重要一步。
