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从牦牛酸奶中分离鉴定产细菌素 XN2 及其细菌素

Isolation and Characterization of Bacteriocin-Producing XN2 from Yak Yoghurt and Its Bacteriocin.

机构信息

College of Food Science, Shanxi Normal University, Taiyuan 030031, China.

Qinba State Key Laboratory of Biological Resources and Ecological Environment, Qinling-Bashan Mountains Bioresources Comprehensive Development C.I.C., Shaanxi Province Key Laboratory of Bio-Resources, College of Bioscience and Bioengineering, Shaanxi University of Technology, Hanzhong 723001, China.

出版信息

Molecules. 2022 Mar 23;27(7):2066. doi: 10.3390/molecules27072066.

Abstract

Lactic acid bacteria (LAB) produce antimicrobial substances that could potentially inhibit the growth of pathogenic and food spoilage microorganisms. XN2, isolated from yak yoghurt, demonstrated antibacterial activity against , , , , , , , , and . The antibacterial activity was estimated to be 3200 AU/mL after 30 h cultivation. Time-kill kinetics curve showed that the semi-purified cell-free supernatants (CFS) of strain XN2 possessed bactericidal activity. Flow cytometry analysis indicated disruption of the sensitive bacteria membrane by semi-purified CFS, which ultimately caused cell death. Interestingly, sub-lethal concentrations of semi-purified CFS were observed to reduce the production of α-haemolysin and biofilm formation. We further investigated the changes in the transcriptional level of gene, which encodes signal molecule synthase (Al-2) induced by semi-purified CFS from strain XN2. In conclusion, XN2 and its bacteriocin showed antagonistic activity at both cellular and quorum sensing (QS) levels. Finally, bacteriocin was further purified by reversed-phase high-performance liquid chromatography (RP-HPLC), named bacteriocin XN2. The amino acid sequence was Met-Lue-Lys-Lys-Phe-Ser-Thr-Ala-Tyr-Val.

摘要

乳酸菌(LAB)产生的抗菌物质可能抑制病原微生物和食物腐败微生物的生长。从牦牛酸奶中分离到的 XN2 对 、 、 、 、 、 、 和 表现出抗菌活性。经过 30 h 培养,抗菌活性估计为 3200 AU/mL。时程杀菌动力学曲线表明,XN2 菌株的无细胞上清液(CFS)具有杀菌活性。流式细胞术分析表明,敏感菌的细胞膜被半纯化 CFS 破坏,最终导致细胞死亡。有趣的是,观察到亚致死浓度的半纯化 CFS 减少了 α-溶血素的产生和生物膜的形成。我们进一步研究了由 XN2 菌株的半纯化 CFS 诱导的信号分子合成酶(Al-2)编码基因 的转录水平的变化。总之,XN2 及其细菌素在细胞和群体感应(QS)水平均表现出拮抗活性。最后,通过反相高效液相色谱(RP-HPLC)进一步纯化细菌素,命名为细菌素 XN2。氨基酸序列为 Met-Lue-Lys-Lys-Phe-Ser-Thr-Ala-Tyr-Val。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7aa5/9000875/f673828e40f2/molecules-27-02066-g001.jpg

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