Identification and characterization of a novel bacteriocin gene cluster in Lysinibacillus boronitolerans.
Identification and characterization of a novel bacteriocin gene cluster in Lysinibacillus boronitolerans.
复制标题
赖氨酸芽孢杆菌中新型细菌素基因簇的鉴定和表征。
DOI：
10.1002/bab.2488
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发表时间：
2023
影响因子：
2.8
通讯作者：
Zhu Liu
中科院分区：
文献类型：
--
作者：
Yanqiong Tang;J. Lei;Xiang Ma;Juanjuan Li;Hong Li;Zhu Liu
抗生素虽然能抑制或杀死病原菌，但滥用抗生素会导致细菌产生耐药性，甚至形成“超级细菌”。“因此，迫切需要探索天然和安全的替代品，如细菌素。本研究通过基因组测序和生物信息学分析，首次预测了耐硼赖氨酸芽孢杆菌的一个未知细菌素基因簇，包括两个生物合成基因、一个调控基因、一个转运相关基因和六个其他基因。随后，在大肠杆菌BL 21中表达了10.24kb的基因簇，并对短小芽孢杆菌、Velezensis芽孢杆菌、Pseudomonaseryningae pv.番茄DC 3000和番茄轴足黄单胞菌致病变种Xanthomonaxonopodispv.木薯属采用70%硫酸铵沉淀法对抗菌物质进行纯化，并采用液相色谱-串联质谱法对抗菌物质进行鉴定。结果表明，该抗菌物质由44个氨基酸组成，与细菌素类似物Piricyclamide 7005 E4 PirE 4的序列同源性为24.1%。通过定点诱变确定了抗菌物质生物合成所需的最小基因集，这表明转录抑制因子和磷酸羟苏氨酸转氨酶都是必不可少的。随后，分析了这两个蛋白在22种赖氨酸芽孢杆菌中的进化和保守性。其中，负责功能的残基进行了鉴定。这些结果为细菌素的生物合成和应用研究奠定了基础。
Although the antibiotics inhibit or kill pathogens, the abuse leads to the resistance formation and even "Super Bacteria." Therefore, it is urgent to explore the natural and safe alternatives such as bacteriocin. In this study, an uncharacterized bacteriocin gene cluster for Lysinibacillus boronitolerans was first predicted by genome sequencing and bioinformatics analysis, of which including two biosynthetic genes, a regulatory gene, a transport-related gene, and six other genes. Subsequently, the 10.24-kb gene cluster was expressed in Escherichia coli BL21, and the lysate effectively inhibited the growths of pathogenic bacteria containing Bacillus pumilus, Bacillus velezensis, Pseudomonas syringae pv. tomato DC3000, and Xanthomonas axonopodis pv. manihotis. The antibacterial substance was purified by 70% ammonium sulfate precipitation and further identified by liquid chromatography-tandem mass spectrometry. The results showed that the antibacterial substance consisted of 44 amino acids and had 24.1% sequence identity with the cyanobacterin Piricyclamide 7005 E4 PirE4, a bacteriocin analogue. The minimal set of genes required for the biosynthesis of the antibacterial substance was determined by site-directed mutagenesis, suggesting both a transcriptional repressor and a phosphohydroxythreonine transaminase were essential. Subsequently, the evolution and conservation of the two proteins were analyzed among 22 Lysinibacillus species. Among them, the residues responsible for functions were identified. Collectively, our results set a solid foundation for investigation of the biosynthesis and application of bacteriocin.
影响因子：
3.7
作者：
Walker BJ;Abeel T;Shea T;Priest M;Abouelliel A;Sakthikumar S;Cuomo CA;Zeng Q;Wortman J;Young SK;Earl AM
通讯作者：
Earl AM