Heterologous Production and Structure Determination of a New Lanthipeptide Sinosporapeptin Using a Cryptic Gene Cluster in an Actinobacterium Sinosporangium siamense

Keita Saito; Keiichiro Mukai; Issara Kaweewan; Hiroyuki Nakagawa; Takeshi Hosaka; Shinya Kodani

doi:10.1007/s12275-023-00059-z

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Heterologous Production and Structure Determination of a New Lanthipeptide Sinosporapeptin Using a Cryptic Gene Cluster in an Actinobacterium Sinosporangium siamense: Keita Saito ¹, Keiichiro Mukai ^2,3, Issara Kaweewan ⁴, Hiroyuki Nakagawa ⁵, Takeshi Hosaka ^2,3, Shinya Kodani ^1,4,6; Journal of Microbiology 2023;61(6):641-648
DOI: https://doi.org/10.1007/s12275-023-00059-z
Published online: June 12, 2023

¹Graduate School of Integrated Science and Technology, Shizuoka University, Shizuoka 422‑8529, Japan, ²Graduate School of Medicine, Science and Technology, Shinshu University, Nagano 399‑4598, Japan, ³Department of Biomolecular Innovation, Institute for Biomedical Sciences, Shinshu University, Nagano 399‑4598, Japan, ⁴Faculty of Agriculture, Shizuoka University, Shizuoka 422‑8529, Japan, ⁵Research Center for Advanced Analysis, Core Technology Research Headquarters, National Agriculture and Food Research Organization (NARO), Ibaraki 305‑8642, Japan, ⁶College of Agriculture, Academic Institute, Shizuoka University, Shizuoka 422‑8529, Japan¹Graduate School of Integrated Science and Technology, Shizuoka University, Shizuoka 422‑8529, Japan, ²Graduate School of Medicine, Science and Technology, Shinshu University, Nagano 399‑4598, Japan, ³Department of Biomolecular Innovation, Institute for Biomedical Sciences, Shinshu University, Nagano 399‑4598, Japan, ⁴Faculty of Agriculture, Shizuoka University, Shizuoka 422‑8529, Japan, ⁵Research Center for Advanced Analysis, Core Technology Research Headquarters, National Agriculture and Food Research Organization (NARO), Ibaraki 305‑8642, Japan, ⁶College of Agriculture, Academic Institute, Shizuoka University, Shizuoka 422‑8529, Japan

Corresponding author: Shinya Kodani ,

Received: 2 February 2023 • Revised: 23 April 2023 • Accepted: 8 May 2023

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Abstract

Lipolanthine is a subclass of lanthipeptide that has the modification of lipid moiety at the N-terminus. A cryptic biosynthetic gene cluster comprising four genes (sinA, sinKC, sinD, and sinE) involved in the biosynthesis of lipolanthine was identified in the genome of an actinobacterium Sinosporangium siamense. Heterologous coexpression of a precursor peptide coding gene sinA and lanthipeptide synthetase coding gene sinKC in the host Escherichia coli strain BL21(DE3) resulted in the synthesis of a new lanthipeptide, sinosporapeptin. It contained unusual amino acids, including one labionin and two dehydrobutyrine residues, as determined using NMR and MS analyses. Another coexpression experiment with two additional genes of decarboxylase (sinD) and N-acetyl transferase (sinE) resulted in the production of a lipolanthine-like modified sinosporapeptin.

Keywords: evolution in vitro;genetic fixation;genetic polymorphism;mutation;varicella vaccine

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Heterologous Production and Structure Determination of a New Lanthipeptide Sinosporapeptin Using a Cryptic Gene Cluster in an Actinobacterium Sinosporangium siamense

J. Microbiol. 2023;61(6):641-648. Published online June 12, 2023

DOI: https://doi.org/10.1007/s12275-023-00059-z

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