摘要

细菌素是由多种细菌产生的具有抗菌活性的核糖体合成肽。从中国传统发酵甘蓝（Suan Tsai）中分离出一株新的细菌素产生菌乳杆菌乳杆菌JLA-9，通过其生理生化特性和16S rDNA序列分析筛选鉴定。使用丁醇提取、凝胶过滤和反相高效液相色谱纯化了一种新的细菌素，命名为植物菌素JLA-9。通过MALDI-TOF-MS分析，得到了1044个Da的植物籽粒JLA-9的分子量。通过MALDI-TOF-MS/MS对植物油苷JLA-9的氨基酸序列进行预测，经埃德曼降解证实。该细菌素对革兰氏阳性菌和革兰氏阴性菌，特别是芽孢杆菌具有广谱抗菌活性。高热稳定性( 20分钟，121℃)和窄pH稳定性( pH 2.0 - 7.0 )。对α-糜蛋白酶、胃蛋白酶、碱性蛋白酶和木瓜蛋白酶敏感。研究了这种细菌素抑制蜡样芽孢杆菌孢子生长的作用方式。植物蛋白JLA - 9在监测孢子的水合、耐热性和2，6 -吡啶二甲酸( DPA )释放方面，在1h内对萌发没有检测到影响。相反，萌发是植物凝集素JLA - 9作用的先决条件。结果表明，植物油杆菌JLA-9在食品工业中对芽孢杆菌的控制具有潜在的应用价值。

Bacteriocins are ribosomally synthesized peptides with antimicrobial activity produced by numerous bacteria. A novel bacteriocin-producing strain, Lactobacillus plantarum JLA-9, isolated from Suan-Tsai, a traditional Chinese fermented cabbage, was screened and identified by its physiobiochemical characteristics and 16S rDNA sequence analysis. A new bacteriocin, designated plantaricin JLA-9, was purified using butanol extraction, gel filtration, and reverse-phase high-performance liquid chromatography. The molecular mass of plantaricin JLA-9 was shown to be 1044 Da by MALDI-TOF-MS analyses. The amino acid sequence of plantaricin JLA-9 was predicted to be FWQKMSFA by MALDI-TOF-MS/MS, which was confirmed by Edman degradation. This bacteriocin exhibited broad-spectrum antibacterial activity against Gram-positive and Gram-negative bacteria, especially Bacillus spp., high thermal stability (20 min, 121 °C), and narrow pH stability (pH 2.0–7.0). It was sensitive to α-chymotrypsin, pepsin, alkaline protease, and papain. The mode of action of this bacteriocin responsible for outgrowth inhibition of Bacillus cereus spores was studied. Plantaricin JLA-9 had no detectable effects on germination initiation over 1 h on monitoring the hydration, heat resistance, and 2,6-pyridinedicarboxylic acid (DPA) release of spores. Rather, germination initiation is a prerequisite for the action of plantaricin JLA-9. Plantaricin JLA-9 inhibited growth by preventing the establishment of oxidative metabolism and disrupting membrane integrity in germinating spores within 2 h. The results suggest that plantaricin JLA-9 has potential applications in the control of Bacillus spp. in the food industry.

https://pubs.acs.org/doi/abs/10.1021/acs.jafc.5b05717