Optimization of FK-506 production in Streptomyces tsukubaensis by modulation of Crp-mediated regulation

FK-506 is a potent immunosuppressive macrocyclic polyketide with growing pharmaceutical interest, produced by Streptomyces tsukubaensis. However, due to low levels synthesized by the wild-type strain, biotechnological production of FK-506 is rather limited. Optimization strategies to enhance the productivity of S. tsukubaensis by means of genetic engineering have been established. In this work primarily global regulatory aspects with respect to the FK-506 biosynthesis have been investigated with the focus on the global Crp (cAMP receptor protein) regulator. In expression analyses and protein-DNA interaction studies, the role of Crp during FK-506 biosynthesis was elucidated. Overexpression of Crp resulted in two-fold enhancement of FK-506 production in S. tsukubaensis under laboratory conditions. Further optimizations using fermentors proved that the strategy described in this study can be transferred to industrial scale, presenting a new approach for biotechnological FK-506 production. Key Points circle The role of the global Crp (cAMP receptor protein) regulator for FK-506 biosynthesis in S. tsukubaensis was demonstrated circle Crp overexpression in S. tsukubaensis was applied as an optimization strategy to enhance FK-506 and FK-520 production resulting in two-fold yield increase

Automated summary

FK-506 is a potent immunosuppressive macrocyclic polyketide produced by Streptomyces tsukubaensis, but its biotechnological production is limited due to low levels synthesized by the wild-type strain. This study investigated the global regulatory aspects of FK-506 biosynthesis, particularly focusing on the role of the Crp regulator. Crp overexpression led to a two-fold increase in FK-506 production, which was successfully transferred to industrial scale using fermentors.