Heterologous production of a new lasso peptide brevunsin in Sphingomonas subterranea
Heterologous production of a new lasso peptide brevunsin in Sphingomonas subterranea
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地下鞘氨醇单胞菌中新套索肽短肽的异源生产
DOI：
10.1007/s10295-018-2077-6
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发表时间：
2018
影响因子：
3.4
通讯作者：
Nakagawa Hiroyuki
中科院分区：
文献类型：
--
作者：
Kodani Shinya;Hemmi Hikaru;Miyake Yuto;Kaweewan Issara;Nakagawa Hiroyuki
以地下鞘氨单胞菌为宿主构建穿梭载体pHSG396Sp进行基因表达。利用PCR扩增了从微小brevundimonas diminuta中获得的一个新的拉索肽生物合成基因簇，并将其整合为表达载体pHSG396Sp-12697L。成功地合成了新的套索肽brevunsin。在地下，含有表达载体，产量高（每L培养10.2 mg）。通过核磁共振（NMR）和质谱（MS/MS）实验确定了brevunsin的化学结构。根据NOE实验获得的信息，确定了brevunsin的三维结构，表明brevunsin具有典型的套索结构。该表达载体系统为细菌基因组编码的未开发套索肽提供了一种新的异源生产方法。
A shuttle vector pHSG396Sp was constructed to perform gene expression usingSphingomonas subterraneaas a host. A new lasso peptide biosynthetic gene cluster, derived fromBrevundimonas diminuta, was amplified by PCR and integrated to afford a expression vector pHSG396Sp-12697L. The new lasso peptide brevunsin was successfully produced byS. 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.