Helmholtz International Lab for Anti-Infectives, Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Shandong University, Qingdao, Shandong, 266237, China.
Microb Biotechnol. 2020 Mar;13(2):397-405. doi: 10.1111/1751-7915.13535.
The Burkholderiales are an emerging source of bioactive natural products. Their genomes contain a large number of cryptic biosynthetic gene clusters (BGCs), indicating great potential for novel structures. However, the lack of genetic tools for the most of Burkholderiales strains restricts the mining of these cryptic BGCs. We previously discovered novel phage recombinases Redαβ7029 from Burkholderiales strain DSM 7029 that could help in efficiently editing several Burkholderiales genomes and established the recombineering genome editing system in Burkholderialse species. Herein, we report the application of this phage recombinase system in another species Paraburkholderia megapolitana DSM 23488, resulting in activation of two silent non-ribosomal peptide synthetase/polyketide synthase BGCs. A novel class of lipopeptide, haereomegapolitanin, was identified through spectroscopic characterization. Haereomegapolitanin A represents an unusual threonine-tagged lipopeptide which is longer than the predicted NRPS assembly line. This recombineering-mediated genome editing system shows great potential for genetic manipulation of more Burkholderiales species to activate silent BGCs for bioactive metabolites discovery.
伯克霍尔德氏菌是具有新兴生物活性天然产物的来源。它们的基因组包含大量隐藏的生物合成基因簇（BGCs），这表明具有新型结构的巨大潜力。然而，由于大多数伯克霍尔德氏菌菌株缺乏遗传工具，限制了对这些隐藏 BGCs 的挖掘。我们之前从伯克霍尔德氏菌菌株 DSM 7029 中发现了新型噬菌体重组酶 Redαβ7029，它可以帮助有效地编辑几种伯克霍尔德氏菌基因组，并在伯克霍尔德氏菌属物种中建立了重组基因组编辑系统。在此，我们报告了该噬菌体重组酶系统在另一种物种 Paraburkholderia megapolitana DSM 23488 中的应用，导致两个沉默的非核糖体肽合成酶/聚酮合酶 BGCs 的激活。通过光谱特征鉴定出一种新型脂肽，haereomegapolitanin。haereomegapolitanin A 代表一种不寻常的苏氨酸标记脂肽，比预测的 NRPS 装配线更长。这种重组介导的基因组编辑系统显示出对更多伯克霍尔德氏菌物种进行遗传操作的巨大潜力，以激活沉默 BGCs 用于生物活性代谢物的发现。
