Degradation of natural polyphenols by methanogenic consortia enriched from digested municipal sludge

Using digested municipal sludge as the inoculum and either rutin, quercetin or hesperidin as the sole external carbon source, methanogenic consortia were enriched which converted various flavonoids at initial concentration of 0.5–3.0 mM during stationary incubation at 37°C in serum bottles with specific rates ranging from 0.025 to 0.073 μmol min−1 (mg protein)−1. In the culture fluid, several hydroxyaromatic metabolites as well as VFA (acetate, propionate, n-butyrate) were detected and biogas was formed in the headspace of the test bottles. Most of these metabolites were identified. Based on their sequential appearance/disappearance in the test cultures it was concluded that following initial hydrolysis of the glycosidic bond by cellular enzymes, ring C of the flavane skeleton was hydrolytically cleaved yielding an A- and B-ring fission product. In case of the flavonol quercetin, phloroglucinol (A-ring) and 3,4-dihydroxyphenylacetate (B-ring) were identified as the fission products whereas the flavanone hesperetin was cleaved with formation of phloroglucinol (A-ring) and 3,4-dihydroxyphenylpropionate (B-ring). In pre-adapted subcultures amended with either hesperidin or hesperetin, all of the formed hydroxyaromatic metabolites disappeared within 100 h of incubation whereas in the culture medium of rutin and quercetin degrading consortia m-cresol (3-methylphenol) was formed as the ultimate hydroxyaromatic metabolite being detectable in considerable amounts even after prolonged incubation.