Enzymatic fractionation of brewer's spent grain and bioconversion of lignin-rich fractions in a colon model in vitro:Dissertation

The objectives of this thesis were to produce lignin-rich fractions from brewer's spent grain (BSG) and to study the interactions of the lignin in these fractions with colon microbiota in vitro. Different milling pre-treatments were studied to enhance enzymatic hydrolysis of BSG carbohydrates. Ball-milling, which was the most efficient treatment, increased carbohydrate solubilisation from 23 to 45 %. Thus, milling notably improved enzymatic solubility of cell wall polysaccharides but was not effective enough to enable their total hydrolysis. Two lignin-rich fractions (24 and 40 % lignin content) were obtained by enzymatic fractionation of BSG using carbohydrases and proteases. In addition, a separate alkaline extraction provided BSG-derived material with low ferulic acid content. BSG and the fractions were used to study if lignin is degraded and metabolised by colon microbiota in a metabolic model and if lignin suppresses microbial conversions in the colon. A number of mono- and dimeric phenolic metabolites were formed upon digestion of BSG and the fractions by the microbiota. It appeared that many of them were structurally lignin-related indicating their release from lignin and conversion by colon microbiota. However, the extent of lignin degradation was estimated to be low. No notable suppression of microbial conversions was detected based on the formation of linear short chain fatty acids. In addition, experiments with pure strains of lactobacilli and bifidobacteria showed no inhibition of growth by a lignin-rich fraction. Association of lignin with carbohydrates or proteinaceous material may have reduced the possible antimicrobial effects of lignin. The results of the present study provide new information on the significance of lignin as part of dietary fibre indicating its partial metabolism by colon microbiota