Peng Jinju, Xie Xingpeng, Fan Tingli, Ma Haotian, Li Yang, Luo Shuaishuai, Yu Mengbo, Ding Yuexia, Ma Yi
College of Coastal Agricultural Sciences, Guangdong Ocean University, Zhanjiang, China.
Department of Agricultural and Animal Husbandry Engineering, Cangzhou Technical College, Cangzhou, China.
Front Pharmacol. 2024 Aug 2;15:1429423. doi: 10.3389/fphar.2024.1429423. eCollection 2024.
The antibacterial protein PAG14 was extracted from a metabolite of G14 isolated from mangrove plants. In this study, , , , and were used as indicator bacteria to screen endophytes that exhibited antibacterial activity. The endophyte culture conditions were optimized to enhance productivity. Subsequently, the culture supernatant was salted using ammonium sulfate, followed by purification using dextran gel chromatography and ion exchange column techniques. Finally, the structures of antibacterial proteins were identified using mass spectrometry. The optimal culture conditions for G14 were 2% mannitol, 0.5% fish peptone, 0.05% KH2PO4 + 0.05% K2HPO4 + 0.025% MnSO4·H2O. The antibacterial substances exhibited stability within the temperature range of 30-40°C and pH range of 5.0-7.0, while displaying sensitivity toward enzymes. The antibacterial activity decreased as the duration of UV irradiation increased. The antibacterial protein PAG14, isolated from the culture broth of G14 through purification using dextran gel and ion-exchange columns, was identified as a class III bacteriocin using LC-MS/MS, similar to Lysozyme C. These findings serve as a theoretical foundation for the investigation and application of bacteriocins in food products.
抗菌蛋白PAG14是从红树林植物中分离出的G14的代谢产物中提取的。在本研究中,使用[具体细菌名称1]、[具体细菌名称2]、[具体细菌名称3]和[具体细菌名称4]作为指示菌来筛选具有抗菌活性的内生菌。对内生菌的培养条件进行了优化以提高产量。随后,用硫酸铵对培养上清液进行盐析,接着使用葡聚糖凝胶色谱和离子交换柱技术进行纯化。最后,用质谱法鉴定抗菌蛋白的结构。G14的最佳培养条件为2%甘露醇、0.5%鱼蛋白胨、0.05% KH2PO4 + 0.05% K2HPO4 + 0.025% MnSO4·H2O。抗菌物质在30 - 40°C的温度范围和5.0 - 7.0的pH范围内表现出稳定性,同时对酶敏感。随着紫外线照射时间的增加,抗菌活性降低。通过葡聚糖凝胶和离子交换柱纯化从G14的培养液中分离出的抗菌蛋白PAG14,使用液相色谱 - 串联质谱法(LC - MS/MS)鉴定为III类细菌素,与溶菌酶C相似。这些发现为细菌素在食品中的研究和应用提供了理论基础。