Engineering <i>Streptomyces coelicolor</i> for heterologous expression of the thiopeptide GE2270A—A cautionary tale

The thiopeptide GE2270A is a clinically relevant, ribosomally synthesised, and post-translationally modified peptide naturally produced by Planobispora rosea. Due to the genetically intractable nature of P. rosea, heterologous expression is considered a possible route to yield improvement. In this study, we focused on improving GE2270A production through heterologous expression of the biosynthetic gene cluster (BGC) in the model organism Streptomyces coelicolor M1146. A statistically significant yield improvement was obtained in the S. coelicolor system through the data-driven rational engineering of the BGC, including the introduction of additional copies of key biosynthetic and regulatory genes. However, despite our best efforts, the highest production level observed in the strains generated in this study is 12× lower than published titres achieved in the natural producer and 50× lower than published titres obtained using Nonomuraea ATCC 39727 as expression host. These results suggest that, while using the most genetically amenable strain as host can be the right choice when exploring different BGC designs, the choice of the most suitable host has a major effect on the achievable yield and should be carefully considered. The analysis of the multiomics data obtained in this study suggests an important role of PbtX in GE2270A biosynthesis and provides insights into the differences in production metabolic profiles between the different strains. One Sentence Summary: Data-driven rational engineering of Streptomyces coelicolor for heterologous production of the thiopeptide antibiotic GE2270A resulted in increased production but encountered unexpected challenges compared to production in the natural producer or the alternative host Nonomuraea ATCC 39727.

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Journal
Journal of Industrial Microbiology & Biotechnology
Published
2024-12-31
DOI
https://doi.org/10.1093/jimb/kuaf019
Primary Topic
Microbial Natural Products and Biosynthesis
Type
article
Field-Weighted Citation Impact
0.00

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article

Engineering Streptomyces coelicolor for heterologous expression of the thiopeptide GE2270A—A cautionary tale

Michelle Rudden, Rosario Pèrez‐Redondo, Antonio Rodríguez‐García, Oksana Bilyk et al.
Journal of Industrial Microbiology & Biotechnology
Microbial Natural Products and Biosynthesis
article

Engineering Streptomyces coelicolor for heterologous expression of the thiopeptide GE2270A—A cautionary tale

Michelle Rudden, Rosario Pèrez‐Redondo, Antonio Rodríguez‐García, Oksana Bilyk, Marianna Iorio, Arianna Tocchetti, Alkisti Manousaki, Suhui Ye, Kamila Schmidt, Erik K. R. Hanko, Olga Genilloud, Emmanuele Severi, Francesco Del Carratore, Dominic R Whittall, Eriko Takano, Stefano Donadio, Rainer Breitling, Gavin H. Thomas, Mercedes Pérez‐Bonilla, Margherita Sosio, Emily J Johnson, Tom Kirkwood
article en

Abstract

The thiopeptide GE2270A is a clinically relevant, ribosomally synthesised, and post-translationally modified peptide naturally produced by Planobispora rosea. Due to the genetically intractable nature of P. rosea, heterologous expression is considered a possible route to yield improvement. In this study, we focused on improving GE2270A production through heterologous expression of the biosynthetic gene cluster (BGC) in the model organism Streptomyces coelicolor M1146. A statistically significant yield improvement was obtained in the S. coelicolor system through the data-driven rational engineering of the BGC, including the introduction of additional copies of key biosynthetic and regulatory genes. However, despite our best efforts, the highest production level observed in the strains generated in this study is 12× lower than published titres achieved in the natural producer and 50× lower than published titres obtained using Nonomuraea ATCC 39727 as expression host. These results suggest that, while using the most genetically amenable strain as host can be the right choice when exploring different BGC designs, the choice of the most suitable host has a major effect on the achievable yield and should be carefully considered. The analysis of the multiomics data obtained in this study suggests an important role of PbtX in GE2270A biosynthesis and provides insights into the differences in production metabolic profiles between the different strains. One Sentence Summary: Data-driven rational engineering of Streptomyces coelicolor for heterologous production of the thiopeptide antibiotic GE2270A resulted in increased production but encountered unexpected challenges compared to production in the natural producer or the alternative host Nonomuraea ATCC 39727.

Journal of Industrial Microbiology & BiotechnologyVol. 52
University of Liverpool (GB), Universidad de Oviedo (ES), University of Manchester (GB), Northumbria University (GB), Fincons Group (Italy) (IT), Junta de Castilla y León (ES), Universidad de León (MX), Fundación Medina (ES), Instituto de Biotecnología de León (ES), University of York (GB), Universidad de León (ES)
UK Research and Innovation, Horizon 2020 Framework Programme, Engineering and Physical Sciences Research Council, Biotechnology and Biological Sciences Research Council
Openalex Percentile: Top 74%
Microbial Natural Products and Biosynthesis
DOIOpen Access PDFOpenAlex
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