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植物乳酸杆菌 GZ1-27 的纯化、特性鉴定及其对蜡样芽孢杆菌作用模式的研究

Purification, Characterization, and Mode of Action of Plantaricin GZ1-27, a Novel Bacteriocin against Bacillus cereus.

机构信息

College of Food Science and Technology , Nanjing Agricultural University , 1 Weigang , Nanjing 210095 , China.

出版信息

J Agric Food Chem. 2018 May 9;66(18):4716-4724. doi: 10.1021/acs.jafc.8b01124. Epub 2018 May 1.

Abstract

Bacillus cereus is an opportunistic pathogen that causes foodborne diseases. We isolated a novel bacteriocin, designated plantaricin GZ1-27, and elucidated its mode of action against B. cereus. Plantaricin GZ1-27 was purified using ammonium sulfate precipitation, gel-filtration chromatography, and RP-HPLC. MALDI-TOF/MS revealed that its molecular mass was 975 Da, and Q-TOF-MS/MS analysis predicted the amino acid sequence as VSGPAGPPGTH. Plantaricin GZ1-27 showed thermostability and pH stability. The antibacterial mechanism was investigated using flow cytometry, confocal laser-scanning microscopy, scanning and transmission electron microscopy, and RT-PCR, which revealed that GZ1-27 increased cell membrane permeability, triggered K leakage and pore formation, damaged cell membrane integrity, altered cell morphology and intracellular organization, and reduced the expression of genes related to cytotoxin production, peptidoglycan synthesis, and cell division. These results suggest that plantaricin GZ1-27 effectively inhibits B. cereus at both the cellular and the molecular levels and is a potential natural food preservative targeting B. cereus.

摘要

蜡样芽胞杆菌是一种机会致病菌,可引起食源性疾病。我们分离到一种新型细菌素,命名为 plantaricin GZ1-27,并阐明了其对蜡样芽胞杆菌的作用模式。plantaricin GZ1-27 采用硫酸铵沉淀、凝胶过滤层析和反相高效液相色谱法进行纯化。MALDI-TOF/MS 显示其分子量为 975 Da,Q-TOF-MS/MS 分析预测其氨基酸序列为 VSGPAGPPGTH。plantaricin GZ1-27 表现出热稳定性和 pH 稳定性。通过流式细胞术、共聚焦激光扫描显微镜、扫描和透射电子显微镜以及 RT-PCR 研究了其抗菌机制,结果表明 GZ1-27 增加了细胞膜通透性,引发 K 渗漏和孔形成,破坏了细胞膜完整性,改变了细胞形态和细胞内组织,并降低了与细胞毒素产生、肽聚糖合成和细胞分裂相关的基因表达。这些结果表明,plantaricin GZ1-27 可有效抑制蜡样芽胞杆菌在细胞和分子水平上的生长,是一种针对蜡样芽胞杆菌的潜在天然食品防腐剂。

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