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Published on: August 1, 2018
A Framework for the In Vivo Production of Extensively Engineered Thiopeptides.
Shinta Ijichi1,2, Shotaro Hoshino1,2, Emiko Nagai2
1Department of Life Science, Faculty of Science, Gakushuin University, 1-5-1 Mejiro, Toshima-Ku, Tokyo 171-8588, Japan.
ACS Synthetic Biology
|July 3, 2026
Summary
Researchers developed a new method for producing engineered lactazole-based thiopeptides in microbes. This framework enables large-scale synthesis of diverse thiopeptide pharmaceuticals with significantly altered macrocycles.
Area of Science:
- Biotechnology
- Synthetic Biology
- Microbial Engineering
Background:
- Thiopeptides, ribosomally synthesized and post-translationally modified peptides, feature complex macrocyclic scaffolds with significant pharmaceutical potential.
- Lactazoles, a subclass of thiopeptides, exhibit versatile biosynthetic pathways suitable for bioengineering diverse macrocyclic peptides.
- Previous in vitro efforts successfully designed novel lactazole-based thiopeptides, but in vivo production of significantly engineered variants remained challenging.
Purpose of the Study:
- To establish a robust framework for the in vivo production of lactazole-based thiopeptides with dramatically engineered macrocycles.
- To investigate the feasibility of microbial systems producing lactazole-based thiopeptides with expanded macrocycles and high amino acid divergence.
- To facilitate the large-scale supply and development of novel thiopeptide-based bioactive molecules.
Main Methods:
- Optimized expression cassettes, focusing on transcriptional terminators like the lazA terminator, for precursor gene expression.
- Evaluated and selected suitable heterologous Streptomyces hosts, identifying Streptomyces sp. TP-A0584 ΔgodA as optimal.
- Implemented optimized culture conditions, including low-temperature cultivation, to enhance thiopeptide production.
Main Results:
- Successfully established a framework enabling in vivo production of lactazole-based thiopeptides.
- Over 90% of tested engineered lactazole-based thiopeptides were produced in microbial systems.
- Achieved mg-scale production, with a maximum titer of 46.4 mg/L, even for variants with up to 73% amino acid divergence in their macrocycles.
Conclusions:
- The developed framework enables efficient in vivo production of highly engineered lactazole-based thiopeptides.
- This breakthrough facilitates the large-scale supply of diverse thiopeptide scaffolds for pharmaceutical development.
- The study paves the way for broader applications of thiopeptide-based bioactive molecules through microbial engineering.
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