The application of the valuable natural product thaxtomin A, a potent bioherbicide from the potato scab pathogenic strains, has been greatly hindered by the low yields from its native producers. Here, we developed an orthogonal transcription system, leveraging extra-cytoplasmic function (ECF) sigma (σ) factor 17 (ECF17) and its cognate promoter P, to express the thaxtomin gene cluster and improve the production of thaxtomin A. The minimal P promoter was determined, and a P promoter library with a wide range of strengths was constructed. Furthermore, a cumate inducible system was developed for precise temporal control of the ECF17 transcription system in ISP5230. Theoretically, the switchable ECF17 transcript... More
The application of the valuable natural product thaxtomin A, a potent bioherbicide from the potato scab pathogenic strains, has been greatly hindered by the low yields from its native producers. Here, we developed an orthogonal transcription system, leveraging extra-cytoplasmic function (ECF) sigma (σ) factor 17 (ECF17) and its cognate promoter P, to express the thaxtomin gene cluster and improve the production of thaxtomin A. The minimal P promoter was determined, and a P promoter library with a wide range of strengths was constructed. Furthermore, a cumate inducible system was developed for precise temporal control of the ECF17 transcription system in ISP5230. Theoretically, the switchable ECF17 transcription system could reduce the unwanted influences from host and alleviate the burdens introduced by overexpression of heterologous genes. The yield of thaxtomin A was significantly improved to 202.1 15.3 g/mL using the switchable ECF17 transcription system for heterologous expression of the thaxtomin gene cluster in ISP5230. Besides, the applicability of this transcription system was also tested in Streptomyces alb J1074, and the titer of thaxtomin A was raised to as high as 239.3 ± 30.6 μg/mL. Therefore, the inducible ECF17 transcription system could serve as a complement of the generally used transcription systems based on strong native constitutive promoters and housekeeping σ factors for the heterologous expression of valuable products in diverse hosts
