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Purification, Characterization, and Mode of Action of Plantaricin GZ1-27, a Novel Bacteriocin against Bacillus cereus
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2018-04-25 00:00:00 , DOI: 10.1021/acs.jafc.8b01124
Hechao Du 1 , Jie Yang 1 , Xiaohong Lu 1 , Zhaoxin Lu 1 , Xiaomei Bie 1 , Haizhen Zhao 1 , Chong Zhang 1 , Fengxia Lu 1
Affiliation  

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.

中文翻译:

新型抗蜡样芽胞杆菌细菌杀菌素Plantarcin GZ1-27的纯化,鉴定和作用方式

蜡状芽孢杆菌是引起食源性疾病的机会病原体。我们分离了一种新型细菌素,命名为plant那霉素GZ1-27,并阐明了其对蜡状芽孢杆菌的作用方式。用硫酸铵沉淀,凝胶过滤色谱和RP-HPLC纯化藤霉素GZ1-27。MALDI-TOF / MS显示其分子量为975 Da,并且Q-TOF-MS / MS分析预测该氨基酸序列为VSGPAGPPGTH。Plantaricin GZ1-27显示出热稳定性和pH稳定性。使用流式细胞仪,共聚焦激光扫描显微镜,扫描和透射电子显微镜以及RT-PCR研究了抗菌机理,结果表明GZ1-27增加了细胞膜通透性,触发了K +渗漏和孔形成,损坏的细胞膜完整性,改变的细胞形态和细胞内组织,以及减少与细胞毒素产生,肽聚糖合成和细胞分裂有关的基因的表达。这些结果表明,plant藤霉素GZ1-27在细胞和分子水平上均有效地抑制蜡状芽孢杆菌,并且是靶向蜡状芽孢杆菌的潜在天然食品防腐剂。
更新日期:2018-04-25
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