Rossi Federico, Cattò Cristina, Mugnai Gianmarco, Villa Federica, Forlani Fabio
Department of Food, Environmental and Nutritional Sciences, Università degli Studi di Milano, 20133 Milano, Italy.
Antioxidants (Basel). 2021 Jun 6;10(6):919. doi: 10.3390/antiox10060919.
The effects of natural compounds on biofilm formation have been extensively studied, with the goal of identifying biofilm formation antagonists at sub-lethal concentrations. Salicylic and cinnamic acids are some examples of these compounds that interact with the quinone oxidoreductase WrbA, a potential biofilm modulator and an antibiofilm compound biomarker. However, WrbA's role in biofilm development is still poorly understood. To investigate the key roles of WrbA in biofilm maturation and oxidative stress, wild-type and ∆A mutant strains were used. Furthermore, we reported the functional validation of WrbA as a molecular target of salicylic and cinnamic acids. The lack of WrbA did not impair planktonic growth, but rather affected the biofilm formation through a mechanism that depends on reactive oxygen species (ROS). The loss of WrbA function resulted in an ROS-sensitive phenotype that showed reductions in biofilm-dwelling cells, biofilm thickness, matrix polysaccharide content, and HO tolerance. Endogenous oxidative events in the mutant strain generated a stressful condition to which the bacterium responded by increasing the catalase activity to compensate for the lack of WrbA. Cinnamic and salicylic acids inhibited the quinone oxidoreductase activity of purified recombinant WrbA. The effects of these antibiofilm molecules on WrbA function was proven for the first time.
天然化合物对生物膜形成的影响已得到广泛研究,目的是在亚致死浓度下鉴定生物膜形成拮抗剂。水杨酸和肉桂酸就是这类化合物的例子,它们与醌氧化还原酶WrbA相互作用,WrbA是一种潜在的生物膜调节剂和抗生物膜化合物生物标志物。然而,WrbA在生物膜发育中的作用仍知之甚少。为了研究WrbA在生物膜成熟和氧化应激中的关键作用,使用了野生型和∆A突变株。此外,我们报道了WrbA作为水杨酸和肉桂酸分子靶点的功能验证。WrbA的缺失并不损害浮游生长,而是通过一种依赖活性氧(ROS)的机制影响生物膜形成。WrbA功能的丧失导致了一种对ROS敏感的表型,表现为生物膜内细胞数量减少、生物膜厚度降低、基质多糖含量减少以及对HO耐受性降低。突变株中的内源性氧化事件产生了一种应激条件,细菌通过增加过氧化氢酶活性来应对这种条件,以弥补WrbA的缺失。肉桂酸和水杨酸抑制了纯化重组WrbA的醌氧化还原酶活性。首次证明了这些抗生物膜分子对WrbA功能的影响。