Heterologous production of a new lasso peptide brevunsin in Sphingomonas subterranea
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A shuttle vector pHSG396Sp was constructed to perform gene expression using Sphingomonas subterranea as a host. A new lasso peptide biosynthetic gene cluster, derived from Brevundimonas diminuta, was amplified by PCR and integrated to afford a expression vector pHSG396Sp-12697L. The new lasso peptide brevunsin was successfully produced by S. subterranea, harboring the expression vector, with a high production yield (10.2 mg from 1 L culture). The chemical structure of brevunsin was established by NMR and MS/MS experiments. Based on the information obtained from the NOE experiment, the three-dimensional structure of brevunsin was determined, which indicated that brevunsin possessed a typical lasso structure. This expression vector system provides a new heterologous production method for unexplored lasso peptides that are encoded by bacterial genomes.
KeywordsLasso peptide Heterologous production Sphingomonas subterranea Brevundimonas diminuta NMR
This study was supported by the Japan Society for the Promotion of Science by Grants-in-aids (Grant number 16K01913). We thank Ms. Tomoko Sato for measurement of MS/MS data.
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Conflict of interest
The authors declare that they have no competing interests.
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