3',4'-Dihydroxycinnamic acid (aka caffeic acid) is a common dietary component found in a variety of plant-derived food products either in a free form or esterified as in chlorogenic acids such as 5-O-caffeoylquinic acid. The dihydroxycinnamate is produced principally by hydrolysis in the colon of 5-O-caffeoylquinic acid and other caffeoylquinic acid esters, and is catabolised by the resident microbiota prior to absorption. In the present study 3',4'-dihydroxycinnamic acid was incubated in vitro, with or without glucose, under anaerobic conditions with faecal slurries obtained from five volunteers. The main resultant catabolites to accumulate were 3-(3',4'-dihydroxyphenyl)propanoic acid (aka dihydrocaffeic acid), 3-(3'-hydroxyphenyl)propanoic acid and phenylacetic acid. Both the rate of degradation of the hydroxycinnamate substrate and the catabolite profile varied between the faecal samples from the individual volunteers. Overall there was no clear cut effect when glucose was added to incubation medium.
The emergence of antibiotic-resistant bacterial pathogens, especially Gram-negative bacteria, has driven investigations into suppressing bacterial virulence via quorum sensing (QS) inhibition strategies instead of bactericidal and bacteriostatic approaches. Here, we investigated several bee products for potential compound(s) that exhibit significant QS inhibitory (QSI) properties at the phenotypic and molecular levels in Chromobacterium violaceum ATCC 12472 as a model organism. Manuka propolis produced the strongest violacein inhibition on C. violaceum lawn agar, while bee pollen had no detectable QSI activity and honey had bactericidal activity. Fractionated manuka propolis (pooled fraction 5 or PF5) exhibited the largest violacein inhibition zone (24.5 ± 2.5 mm) at 1 mg dry weight per disc. In C. violaceum liquid cultures, at least 450 µg/ml of manuka propolis PF5 completely inhibited violacein production. Gene expression studies of the vioABCDE operon, involved in violacein biosynthesis, showed significant (≥two-fold) down-regulation of vioA, vioD and vioE in response to manuka propolis PF5. A potential QSI compound identified in manuka propolis PF5 is a hydroxycinnamic acid-derivative, isoprenyl caffeate, with a [M-H] of 247. Complete violacein inhibition in C. violaceum liquid cultures was achieved with at least 50 µg/ml of commercial isoprenyl caffeate. In silico docking experiments suggest that isoprenyl caffeate may act as an inhibitor of the violacein biosynthetic pathway by acting as a competitor for the FAD-binding pockets of VioD and VioA. Further studies on these compounds are warranted toward the development of anti-pathogenic drugs as adjuvants to conventional antibiotic treatments, especially in antibiotic-resistant bacterial infections.