Targeted and Nontargeted Metabolomics of Amino Acids and Bioactive Metabolites in Probiotic-Fermented Unhopped Beers Using Liquid Chromatography High-Resolution Mass Spectrometry

Abstract

Beer is one of the most popular beverages in the world. The increased popularity of craft beers has led to the development of unique beers that are alcohol-free, gluten-free, low calorie, or with functional properties through fermentation with probiotic microorganisms. In this study, functional unhopped beers were evaluated by utilizing probiotics (Lacticaseibacillus paracasei Lpc-37 and ibSium Saccharomyces cerevisiae CNCM I-3856) as starter cultures. The metabolites produced by probiotics were investigated using a nontargeted metabolomics approach and identified against metabolomics databases (Kyoto Encyclopedia of Genes and Genomes (KEGG), Human Metabolome Database (HMDB), Yeast Metabolome Database (YMDB), METLIN tandem mass spectrometry (MS/MS)). Derivatives of branched-chain (leucine) and aromatic amino acids (phenylalanine, tryptophan, and tyrosine) were enriched (one-way analysis of variance (ANOVA) p < 0.05) in probiotic-fermented unhopped beers, especially tryptophan metabolites. In addition, the synergistic effects of yeast−lactic acid bacteria (LAB) interactions led to further enrichment of higher acids such as (S)-(−)-2-hydroxyisocaproic acid, phenyllactic acid, hydroxyphenyllactic acid, and indolelactic acid. The potential pathways for the formation of novel bioactive tryptophan metabolites (indole and indoleacrylic acid) by LAB were elucidated. Altogether, probiotic LAB-fermented unhopped beer showed the highest antioxidant capacity and total phenolic content. This work provides the basis for the discovery of bioactive metabolites in probiotic-fermented foods.

Metabolomic analysis of the effects of a mixed culture of Saccharomyces cerevisiae and Lactiplantibacillus plantarum on the physicochemical and quality characteristics of apple cider vinegar

“…DPPH radical scavenging capacity was determined as described previously ( 21 , 22 ). The ABTS radical scavenging capacity was determined as described previously ( 23 , 24 ), with minor modifications.…”

Fungal–Bacterial Mutualism: Species and Strain-Dependent Simultaneous Modulation of Branched-Chain Esters and Indole Derivatives in Fermented Sausages through Metabolite Cross-Feeding

“…22 Furthermore, targeted and nontargeted metabolomics analyses have revealed that synergistic effects of yeast−bacteria interactions led to further enrichment of amino acids derivatives in beer fermentation. 23 Thus, the interaction of D. hansenii with CNS and LAB may promote the accumulation of branched-chain esters and indole derivatives, respectively.…”

“…hansenii in fermented sausages . Furthermore, targeted and nontargeted metabolomics analyses have revealed that synergistic effects of yeast–bacteria interactions led to further enrichment of amino acids derivatives in beer fermentation . Thus, the interaction of D.…”

A novel screening method for free non-standard amino acids in human plasma samples using AccQ·Tag reagents and LC-MS/MS

“…10 In addition, the use of AQC derivatization has been extended to the analysis of non-standard amino acids, metabolites with amine moieties, and N -glycans. 12–14 Due to the flexibility of the detection device coupled to the LC, high sensitivity, good chromatographic separation, and high selectivity, AQC derivatization has been used in a wide range of application areas including the analysis of neurotransmitter amino acids, 15 characterizing the nutritional value of food and beverage, 16,17 and even soil amino acid analysis. 18…”

“…10 In addition, the use of AQC derivatization has been extended to the analysis of non-standard amino acids, metabolites with amine moieties, and N-glycans. [12][13][14] Due to the exibility of the detection device coupled to the LC, high sensitivity, good chromatographic separation, and high selectivity, AQC derivatization has been used in a wide range of application areas including the analysis of neurotransmitter amino acids, 15 characterizing the nutritional value of food and beverage, 16,17 and even soil amino acid analysis. 18 Aer AQC derivatization, the hydrophobic tag allows good chromatographic retention and separation of amino acids with different polarities on a reverse phase C18 column under generic chromatography conditions thereby enabling easy LC method development.…”