Identification and application of a strong bidirectional acmN2p promoter from actinomycin D-producing streptomycetes

Abstract

Natural product biosynthesis is controlled at multiple levels. Characterization of naturally occurring promoters has facilitated the study of the synthetic biology of natural products. Herein, we report the discovery of two high-yield actinomycin D (ActD)-producing streptomycetes and the identification of a strong bidirectional acmN2p promoter from the ActD gene clusters and its application in heterologous expression of three core genes involved in the bacterial alkaloid bohemamine biosynthesis, providing a good example for identification of new promoters for synthetic biological applications.

Keywords

Actinomycin D / Bidirectional promoter / Bohemamine / Streptomyces

Conclusion

In conclusion, natural BDP acmN2p was identified in two ActD-producing Streptomyces strains and was found to drive the expression of tsr^r and kan^r in three model Streptomyces hosts. The acmN2p BDP was further demonstrated to drive the expression of three genes from the bohemamine BGC in heterologous Streptomyces hosts, which led to the production of bohemamine analogs 2 and 3. This study may inspire the discovery of additional BDPs from natural BGCs for synthetic biological applications.

Data Availability Statement

All data generated or analyzed during this study are included in this published article and its supplementary information files or are available upon request.

Declaration of Competing Interest

No potential conflict of interest was reported by the authors.

CRediT authorship contribution statement

Sainan Li: Formal analysis, Investigation, Methodology, Writing - original draft. Danfeng Tang: Funding acquisition, Investigation, Methodology, Writing - original draft. Xu Zhao: Investigation, Methodology. Manxiang Zhu: Formal analysis, Writing - original draft. Xiangcheng Zhu: Formal analysis. Yanwen Duan: Funding acquisition, Supervision. Yong Huang: Conceptualization, Funding acquisition, Supervision, Writing - review & editing.

Acknowledgments

This work was supported in parts by National Natural Science Foundation of China grants (82173688 and 82373772), The science and technology innovation Program of Hunan Province (2021RC4067), and a research fund from Institute of Health and Medicine (IHM), Hefei Comprehensive National Science Center (to Y.H.), the Chinese Ministry of Education 111 Project (BP0820034) (to Y.D.), and the Fundamental Research Funds for the Central Universities of Central South University 2020zzts247 (to D.T.). We thank the core facility of IHM for FACS analysis.

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