Kong Junhao, Wang Yanan, Xia Kai, Zang Ning, Zhang Hong, Liang Xinle
School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou, 310018 China.
Institute of Food Biotechnology, Zhejiang Gongshang University, Hangzhou, 310018 China.
3 Biotech. 2021 Feb;11(2):97. doi: 10.1007/s13205-021-02663-5. Epub 2021 Jan 27.
This study aimed to investigate the anti-quorum sensing (QS) activity of leaf extracts (AALE) towards PAO1 as well as the underlying molecular mechanisms. Using a biosensor CV026, AALE were found to have anti-QS activity as AALE treatment significantly inhibited the violacein production of CV026 while produced little effect on the cell growth. Beyond that a higher dosage of AALE inhibited cell growth, sub-MIC of AALE significantly reduced the production of QS-regulated virulence factors (pyocyanin, elastase, and rhamnolipid), biofilm formation, and the swarming and swimming motility in PAO1 with a dosage-dependent manner. Quantitative real-time PCR (qRT-PCR) analysis did not detect the direct inhibitory effect of AALE on the expression of QS genes (, , , and ). By iTRAQ-based quantitative proteomic analysis, 129 proteins were found to be differentially expressed upon AALE treatment, with 85 upregulated and 44 downregulated proteins, respectively. Functional enrichment analysis of the differential proteins revealed that AALE exerted anti-QS activity towards PAO1 by upregulating the expression of the global regulator CsrA, inducing oxidative stress, and perturbing protein homeostasis. Moreover, the inhibitory effect of AALE on the virulence of PAO1 was likely to be achieved by attenuating the expression of QS-regulated genes instead of QS genes. Collectively, the results of this study provide a basis for the future use of AALE as a preservative in controlling food spoilage caused by .
The online version contains supplementary material available at 10.1007/s13205-021-02663-5.
本研究旨在探究叶片提取物（AALE）对铜绿假单胞菌PAO1的群体感应（QS）抑制活性及其潜在分子机制。使用生物传感器CV026，发现AALE具有抗QS活性，因为AALE处理显著抑制了CV026的紫色杆菌素产生，而对细胞生长影响较小。除此之外，较高剂量的AALE抑制细胞生长，AALE的亚抑菌浓度以剂量依赖方式显著降低PAO1中QS调控的毒力因子（绿脓菌素、弹性蛋白酶和鼠李糖脂）的产生、生物膜形成以及群集和游动运动性。定量实时PCR（qRT-PCR）分析未检测到AALE对QS基因（lasI、lasR、rhlI和rhlR）表达的直接抑制作用。通过基于iTRAQ的定量蛋白质组学分析，发现129种蛋白质在AALE处理后差异表达，分别有85种上调蛋白和44种下调蛋白。差异蛋白的功能富集分析表明，AALE通过上调全局调节因子CsrA的表达、诱导氧化应激和扰乱蛋白质稳态对PAO1发挥抗QS活性。此外，AALE对PAO1毒力的抑制作用可能是通过减弱QS调控基因而非QS基因的表达来实现的。总体而言，本研究结果为未来将AALE用作防腐剂来控制由PAO1引起的食品腐败提供了依据。
在线版本包含可在10.1007/s13205-021-02663-5获取的补充材料。
