Recently, we reported that a novel phenolic compound isolated from Cedrus deodara, 3-p-trans-coumaroyl-2-hydroxyquinic acid (CHQA), exhibits a potent antioxidant activity. The present study aimed to evaluate the antibacterial activity of CHQA against eleven food-borne pathogens and to elucidate its mechanism of action against Staphylococcus aureus. The results from minimum inhibitory concentration (MIC) determinations showed that CHQA exhibited moderate inhibitory effects on all of the tested pathogens with MIC values ranging from 2.5–10 mg/mL. Membrane potential measurements and flow cytometric analysis demonstrated that CHQA damaged the cytoplasmic membrane of S. aureus, causing a significant membrane hyperpolarization with a loss of membrane integrity. Moreover, CHQA induced an increase in membrane fluidity and conformational changes in membrane protein of S. aureus, suggesting that CHQA probably acts on the cell membrane by interactions with membrane lipid and protein. Transmission electron microscopic observations further confirmed that CHQA disrupted the cell membrane of S. aureus and caused severe morphological changes, which even led to leakage of intracellular constituents. These findings indicated that CHQA could have the potential to serve as a natural antibacterial agent to control and prevent the growth of pathogens in food and in food-processing environments.

Shikimic acid (SA) has been reported to possess antibacterial activity against Staphylococcus aureus, whereas the mode of action of SA is still elusive. In this study, the antibacterial activity and mechanism of SA toward S. aureus by cell membrane damage was investigated. After SA treatment, massive K⁺ and nucleotide leakage from S. aureus, and a significant change in the membrane potential was observed, suggesting SA may act on the membrane by destroying the cell membrane permeability. Through transmission electron microscopic observations we further confirmed that SA can disrupt the cell membrane and membrane integrity. Meanwhile, SA was found to be capable of reducing the membrane fluidity of the S. aureus cell. Moreover, the fluorescence experiments indicated that SA could quench fluorescence of Phe residues of the membrane proteins, thus demonstrating that SA can bind to S. aureus membrane proteins. Therefore, these results showed the antibacterial activity of SA against S. aureus could be caused by the interactions of SA with S. aureus membrane proteins and lipids, resulting in causing cell membrane dysfunction and bacterial damage or even death. This study reveals the potential use of SA as an antibacterial agent.

Recently, studies on theabrownins (TBs), the main bioactive polymeric pigments found in dark tea, have received increasing attention for its health effects. Thus far, information on their structural characteristics is unclear. In the present study, theabrownins were isolated from single species Eurotium cristatum PW-1-fermented loose tea and their structural and hypolipidemic characteristics were studied for the first time. The theabrownins were fractionated by their molecular weights and were then analyzed. Ultraviolet–visible spectrophotometry (UV-Vis) and Flourier transformation infrared spectroscopy (FT-IR) showed that they were polymerized phenolic substances containing abundant hydroxy and carboxyl groups. All theabrownin samples exhibited hypolipidemic activity in high-fat zebrafish; among which TBs-10-30k sample, decreased lipid level in high-fat zebrafish to 51.57% at 1000 μg/mL, was most effective. It was found that TBs-10-30k was a type of amorphous and thermostable polymer with slice shape and smooth surface under scanning electron microscope (SEM). Atomic force microscope (AFM) analysis showed that it had island-like structure because of aggregation of theabrownin molecules. Pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) analysis further showed that the main pyrolytic products of TBs-10-30k were hexadecanoic acid (33.72%), phenol (14.90%), and eicosane (12.95%), indicating TBs-10-30k was mainly composed of phenols, lipids, saccharides, and proteins. These results not only facilitate subsequent identification of theabrownins, but also provide insights into the applications of theabrownins in functional foods.

Aspergillus niger is one of the dominant microorganisms presented in dark tea fermentation. In this study, the biotransformation of steamed green tea leaves fermented by A. niger PW-2 was characterized using metabolomic and proteomic approaches. We observed that, after fermentation, the contents of volatile compounds contributing to the “green” aroma, including linalool, L-α-terpineol and geraniol, decreased significantly. Meanwhile, the astringency taste and contents of metabolites contributing to the taste (catechins) reduced significantly during fermentation. Additionally, the contents of theabrownins, which have health benefits, obviously increased. The bitter and umami tastes were also changed due to the variations in bitter-taste and umami-taste amino acids. We also found that glycoside hydrolases, tannases, catechol oxidases, peroxidases and laccases secreted by A. niger PW-2 were responsible for the metabolism of phenolic compounds and their derivatives (theaflavins, thearubingins and theabrownins). Finally, the metabolic pathways involved in the biosynthesis and degradation of characteristic metabolites were found to reveal the biotransformation characteristics of dark tea fermented with A. niger PW-2.

The biofilm formation of Staphylococcus aureus on food contact surfaces is the main risk of food contamination. In the present study, we firstly investigated the inhibitory effect of quinic acid (QA) on biofilm formed by S. aureus. Crystal violet staining assay and microscopy analysis clearly showed that QA at sub-MIC concentrations was able to significantly reduce the biofilm biomass and cause a collapse on biofilm architecture. Meanwhile, fibrinogen binding assay showed that QA had obviously effect on the S. aureus bacteria adhesion. XTT reduction assay and confocal laser scanning microscopic images revealed that QA significantly decreased metabolic activity and viability of biofilm cells. In addition, qRT-PCR analysis explored the potential inhibitory mechanism of QA against biofilm formation, which indicated that QA significantly repressed the gene sarA and activated the gene agrA. Moreover, QA exhibited a highly ability to reduce the number of sessile S. aureus cells adhered on the stainless steel. So, it was suggested that QA could be used as a promising antibiofilm agent to control biofilm formation of S. aureus.