Abstract
The thermophilic bacterium Thermosporothrix hazakensis belongs to a class of Ktedonobacteria in the phylum Chloroflexota. Lanthipeptides are a naturally occurring peptide group that contains antibacterial compounds such as nisin. To find a new lanthipeptide that is a possible candidate for an antibacterial reagent, we performed genome-mining of T. hazakensis and heterologous expression experiments. Based on genome-mining, the presence of a total of ten putative biosynthetic gene clusters for class I and class II lanthipeptides was indicated from the genome sequence of T. hazakensis. New lanthipeptides named hazakensins A and B were produced by heterologous expression of a class I lanthipeptide biosynthetic gene cluster in the expression host Escherichia coli. Co-expression of the biosynthetic gene cluster with tRNA-Glu and glutamyl-tRNA synthetase coding genes derived from T. hazakensis increased the production yield of both lanthipeptides by about 4–6 times. The chemical structures of hazakensins A and B including the bridging pattern of lanthionine/methyllanthionine rings were determined by NMR and MS experiments. Since production of hazakensins A and B was not observed in the native strain T. hazakensis, heterologous production was an effective method to obtain the lanthipeptides derived from the biosynthetic gene cluster. This is the first report of heterologous production of class I lanthipeptides originating from the filamentous green non-sulfur bacteria, to the best of our knowledge. The success of heterologous production of hazakensins may lead to the discovery and development of new lanthipeptides derived from the origins of bacteria in the phylum Chloroflexota.
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Acknowledgements
We thank Ms. Tomoko Sato for the measurement of MS data.
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This study was supported by Grants-in-Aid for Scientific Research: JSPS KAKENHI Grant Number 20K05848 and 21F21095, Institute for Fermentation, OSAKA (IFO): Grant Number G-2021-3-010, and Kobayashi Foundation: Grant Number 196. I. K. was supported by JSPS Postdoctoral Fellowships for Research in Japan (Standard Program).
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IK carried out heterologous production and structure determination of peptides. SI performed molecular cloning. HN performed analysis using NMR and ESI-MS. IK and SK conceived the study and wrote the manuscript.
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Kaweewan, I., Ijichi, S., Nakagawa, H. et al. Heterologous production of new lanthipeptides hazakensins A and B using a cryptic gene cluster of the thermophilic bacterium Thermosporothrix hazakensis. World J Microbiol Biotechnol 39, 30 (2023). https://doi.org/10.1007/s11274-022-03463-6
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DOI: https://doi.org/10.1007/s11274-022-03463-6