Heterologous production of a new lasso peptide brevunsin in Sphingomonas subterranea

Kodani, Shinya; Hemmi, Hikaru; Miyake, Yuto; Kaweewan, Issara; Nakagawa Hiroyuki,

Heterologous production of a new lasso peptide brevunsin in Sphingomonas subterranea

2018

Kodani, Shinya | Hemmi, Hikaru | Miyake, Yuto | Kaweewan, Issara | Nakagawa Hiroyuki,

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.

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