Exploring the role of the major gut microbiota clusters on nutritional and functional benefits of nutrients and non-nutrients
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1 Exploring the role of the major gut microbiota clusters on nutritional and functional benefits of nutrients and non-nutrients Ian Rowland Department of Food & Nutritional Sciences University of Reading, UK
2 Conflict of Interest Disclosure This work was conducted by an expert group of the European branch of the Interna9onal Life Sciences Ins9tute, ILSI Europe. The non- industry members within this expert group were offered support for travel and accommoda9on costs to acend mee9ngs for discussing the manuscript, and a small compensatory sum (honorarium) with the op9on to decline. 2
3 ILSI Europe group: Role of microbiota on nutritional & functional benefits of nutrients & non-nutrients Aims of the expert group Evaluate role of microbiota in metabolism of dietary compounds Review mechanisms and pathways involved Identify main types of microorganisms involved Consider the methodologies for investigating gut microbiota metabolism Outputs Shortt et al Systematic review of the effects of the intestinal microbiota on selected nutrients and non-nutrients Eur J Nutr Rowland et al Gut microbiota functions: metabolism of nutrients and other food components Eur J Nutr Apr 9. 3
4 Human gut microbiota phyla & genera Firmicutes 4 Clostridium Roseburia Faecalibacterium Blautia Dorea Lactobacillus Peptostreptococcus Eubacterium Streptococcus Staphylococus Butyrivibrio Bacteroidetes Bacteroides Prevotella Verrucomicrobia Akkermansia Proteobacteria Escherichia Klebsiella Desulfovibrio Actinobacteria Bifidobacterium Collinsella 90% of bacteria are in Bacteroidetes/ Firmicutes phyla 4
5 Gut microbiota - metabolism Ø Large metabolic potential - equivalent to, but different from that of liver Ø Microbiota metabolic range Reduction Demethylation Hydrolysis Deamination Polysaccharide Nitrosation fermentation Ring fission Dehydroxylation Aromatization Methylation Oligomer breakdown 5 5
6 Dietary components reviewed Carbohydrates Energy homeostasis Proteins Lipids Bile acids Vitamins Phytochemicals/polyphenols 6
7 Carbohydrate fermenta7on Polysaccharides Oligosaccharides Mucins NSP/Resistant starch Microbial fermenta9on H 2 CO 2 Short chain faay acids (SCFA) CH 4 Acetate Propionate Butyrate Lactate Methanobrevibacter smithii H 2 S Desulfovibrio spp Most gut organisms Eubacterium hallii Roseburia Coprococcus catus Megasphera elsdenii Veillonella spp Ruminococcus obeum Bacteroidetes Eubacterium rectale Eubacterium hallii Roseburia spp Coprococcus spp Faecalibacterium prausnitzii Bifidobacterium spp Lactobacillus spp 7
8 Energy homeostasis Studies in humans and animals suggest microbiota is involved in assimilation, storage & expenditure of energy obtained from dietary substrates. Some studies report higher Firmicutes/Bacteroides ratio in obese subjects Higher BMI associated with increased abundance of Eubact. ventrosium and Roseburia intestinalis, E. rectale Weight-loss regimes (diet or gastrectomy) associated with Bacteroidetes Firmicutes inc Clostridium, Eubacterium, Coprococcus). Potential mechanisms Gut microbes break down non-digestible carbohydrates to SCFA allowing the host to salvage energy from indigestible dietary substrates. SCFA energy intake : Pr & Bu increase the levels of satiety hormones PYY and GLP-1, Ac & Pr increase expression of leptin Role of specific gut microbes and SCFA in energy balance remains to be clarified 8
9 Protein metabolism clostridia peptostreptococci peptococci Bacteroides spp Eubacterium hallii Clos barlettii Branched chain fatty acids (BCFA) - Isobutyrate (from val) - 2-methylbutyrate (i-leu) - Isovalerate (leu) p- cresol Davila et al. Pharmacological Res.69:
10 Lipid metabolism Linoleic acid reduced to stearic acid (via CLA and vaccinic acid) in vitro by range of gut bacteria Phosphatidylcholine to TMA 10
11 Microbiota metabolism of choline to TMA and TMAO TMA produced by solely by gut microbiota High-fat diets lead to increased production of TMA and TMAO, - risk factors for cardiovascular disease Choline metabolizers: Proteobacteria : Desulfovibrio desulfuricans, Klebsiella spp, Escherichia spp, Edwardsiella tarda, Proteus penneri Firmicutes: Clos sporogenes, hathewayi, asparagiforme Martinez-del-Campo mbio 6(2) e00042 (2015) Romano et al mbio 6(2) e02481 (2015) 11
12 Bile acids 1. Deconjugation by bacterial bile salt hydrolases (BSH) ç Bile salts BSH ç Bile acids Bile salt hydrolase BSH genes identified in the main bacterial genera including Bacteroides, Bifidobacterium, Clostridium, Lactobacillus, and Listeria Most hydrolyze both glyco and tauro-conjugates. Reduces toxicity of bile acids, releases N, S and C atoms 7α-dehydroxylation Deconjugation reduces efficiency of BA for emulsifying lipids and micelle formation 12
13 Bile acids further metabolism DCA & LA are more cytotoxic and genotoxic, to colon mucosa than CA & CDA 7α-dehydroxylation Epimerization ç Secondary bile acids 7-α-dehydroxylase main bacterial genera include Clostridium, Eubacterium Epimerization Ursodeoxycholic acid Main genera : Bacteroides, Clostridium, Egghertella, Peptostreptococcus, Ruminococcus, Eubacterium 13
14 Vitamin synthesis Vitamin synthesis genes common esp. vitamin K and B group vitamins - biotin, cobalamin, folate, nicotinic acid, panthothenic acid, pyridoxine, riboflavin and thiamine For riboflavin & biotin, all tested microbes in Bacteroidetes Fusobacteria and Proteobacteria phyla had required pathways, fewer Firmicutes and Actinobacteria had the pathways Bacteroidetes is most important phyla for vitamin synthesis Many of these vitamins are utilized by other bacteria 14
15 Vitamin synthesis 15 Vitamin Intracellular concentra7on [mmol/gdw] Dietary reference intake [mg/ day] Bio9n 9.0 x Cobalamin 8.5 x Folate 5.0 x Niacin 3.3 x Pantothenate 2.3 x Pyridoxine 5.8 x Riboflavin 9.0 x Thiamin 8.7 x %DRI from gut microbiota MagnusdoYr et al. Front Genet. 2015
16 Phytochemicals Polyphenols 16
17 Polyphenols Often poorly absorbed in small intestine è colon Parent polyphenols are extensively metabolized by the microbiota, (deglycosylation, ring fission, dehydroxylation) - can impact bioactivity Metabolism often requires consortia or 2 or more microbes Large interindividual variations in absorption and excretion ascribed to differences in gut microbiota 17
18 Pathways of colonic degradation of the flavonoid rutin OH OH OH HO OH O O O-Rutinose OH Deglycosylation HO OH O O OH Quercetin Further degradation HO COOH Rutin Dehydroxylation HO HO COOH 3,4-dihydroxyphenylacetic acid Ring fission, water elimination, dehydroxylation HO HO Protocatechuic acid 18 COOH HO 3-hydroxyphenylacetic acid Absorption from the colon COOH 3-(3-hydroxyphenyl)- propionic acid HO β-oxidation NH CO COOH + glycination 3-hydroxyhippuric acid
19 Gut microbiota & inter-individual variation in polyphenol metabolism Differences in composition of microbiota between individuals can have significant effects on extent of metabolism and metabolite profile Examples: Isoflavonoids (daidzein to equol) Naringin Anthocyanins Lignans Tea catechins Rutin 19
20 Isoflavone metabolism by gut bacteria Lactobacillus mucosae + Enterococcus faecium + Finegoldia magna + Veillonella sp Clostridium spp. Eubacterium ramulus Slakia isoflavoniconvertens, Slakia equolifaciens, Adlercreutzia equolifaciens Blaut et al 2003; Decroos et al 2005; MaChies et al 2009, 2012) 20
21 Equol excretion in subjects consuming soy isoflavonoids Subjects consumed soy burgers (56mg IF) - 1/day for 17days (Rowland et al 2003) 21
22 Methodologies Isolated cultures Gut microbial enzyme activity Omics approaches Metagenomics Metatranscriptomics Metaproteomics Metabolomics Mathematical modeling 22
23 Omics approaches for studying gut microbial metabolism/function Metagenomics study functional genes associated with specific microbial types, Meta-transcriptomics monitor active bacteria, reveals functional roles (eg CHO metabolism) info on functional dysbiosis, Meta-proteomics confirming microbial function (faecal meta proteome is subject-specific and stable) Metabonomics pathway analysis, metabolic biomarkers of disease risk 23
24 Conclusions Gut microbiota metabolism enlarges the capacity of host to metabolize range of dietary components and extends the range of metabolites formed CHO metabolism is major function of microbiota pathways and microbes well studied. Microbial metabolites of nutrients and non-nutrients can be important cell signaling molecules (SCFA, bile acids) and have impacts on health (SCFA, TMA, phenolics) Large interindividual variation in microbiota consequences for metabolism of dietary compounds and health Omics provide insight into microbiota function at high resolution 24
25 Acknowledgements Members of the ILSI Europe working group Jean-Michel Antoine (Danone) Glenn Gibson (University of Reading) Oliver Hasselwander (DuPont) Alexandra Meynier (Mondelez) Arjen Nauta (Royal Friesland Campina) Karen Scott (Rowett Institute) Colette Short (Johnson & Johnson) Jonathan Swan (Imperial College London) Ines Thiele (University of Luxembourg) Kieran Tuohy (Fondazione E Mach) Jessica Türck (Yakult Europe) Joan Vermeiren (Cargill) Estefania Noriega (ILSI Europe) Peter Putz (ILSI Europe) Tobias Recker (ILSI Europe) 25
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