Helmholtz International Lab for Anti-Infectives, Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, Shandong University, Qingdao 266237, China.
Jiangsu Marine Resources Development Research Institute, Lianyungang 222005, China.
Molecules. 2021 Jan 29;26(3):700. doi: 10.3390/molecules26030700.
The genus possesses ecological and metabolic diversities. A large number of silent biosynthetic gene clusters (BGCs) in the genome remain uncharacterized and represent a promising resource for new natural product discovery. However, exploitation of the metabolomic potential of is limited by the absence of efficient genetic manipulation tools. Here, we screened a bacteriophage recombinase system Redγ-BAS, which was functional for genome modification in the plant pathogen ATCC 10248. By using this recombineering tool, the constitutive promoters were precisely inserted in the genome, leading to activation of two silent nonribosomal peptide synthetase gene clusters ( and ) and production of corresponding new classes of lipopeptides, burriogladiodins A-H (-) and haereogladiodins A-B (-). Structure elucidation revealed an unnatural amino acid - dehydrobutyrine (Dhb) in - and an -Dhb in -. Notably, compounds - and feature an unusual threonine tag that is longer than the predicted collinearity assembly lines. The structural diversity of burriogladiodins was derived from the relaxed substrate specificity of the fifth adenylation domain as well as chain termination conducted by water or threonine. The recombinase-mediating genome editing system is not only applicable in but also possesses great potential for mining meaningful silent gene clusters from other species.
该属具有生态和代谢多样性。大量未被表征的沉默生物合成基因簇（BGCs）存在于基因组中，代表了新天然产物发现的有前途的资源。然而，由于缺乏有效的遗传操作工具，对 的代谢组学潜力的开发受到限制。在这里，我们筛选了一种噬菌体重组酶系统 Redγ-BAS，该系统在植物病原体 ATCC 10248 中可用于基因组修饰。通过使用这种重组工具，精确地将组成型启动子插入基因组中，导致两个沉默的非核糖体肽合成酶基因簇（和）被激活，并产生相应的新类脂肽，即 burriogladiodins A-H（-）和 haereogladiodins A-B（-）。结构阐明揭示了 -和 -中的非天然氨基酸 -脱氢丁氨酸（Dhb）。值得注意的是，化合物 -和 具有不寻常的苏氨酸标签，比预测的共线性组装线长。burriogladiodins 的结构多样性源于第五个腺苷酸化结构域的宽松底物特异性以及由水或苏氨酸介导的链终止。重组酶介导的基因组编辑系统不仅适用于 ，而且对于从其他 物种中挖掘有意义的沉默基因簇也具有巨大潜力。
