Fermentation process of apple juice investigated by NMR spectroscopy

Cider is a slightly alcoholic beverage and is an important and promising segment of the fruit industry. In this study, a still cider produced by fermentation of apples juice was investigated. A simultaneous characterization of bioactive compounds in apple juice and the intermediate products of the cidermaking process was performed for the first time using Nuclear Magnetic Resonance spectroscopy. Alongside sugar consumption, a progressive increase in trigonelline, chalcone, fumarate, caffeic acid, uracil, tyrosol and xanthine content was detected during the alcoholic fermentation process. Of note was the relatively high tyrosol content which imparts health benefits in the cider. The concomitant changes in phenol, amino acid and organic acid (malic, lactic, quinic, pyruvic, citric and succinic) content, was important when monitoring the fermentation process and assessing quality control of the final product

NMR-based metabolic profiling of different yeast fermented apple juices

NMR based metabolic profiling was used to investigate metabolic changes in ciders fermented by yeast species as valuable tool for the identification of metabolites responsible for yeast activities. 1H NMR spectra of apple ciders produced by Saccharomyces cerevisiae EC1118, Torulaspora delbrueckii TD291, Starmerella bacillaris YR21, Hanseniaspora osmophila HO16, Hanseniaspora uvarum Y4M, and Saccharomyces uvarum SU3, revealed to be dominated by glycerol and organic acids signals. The aromatic region showed the presence of low intense signals referred to epicatechin, catechin, chlorogenate, xanthine, uracil, tyrosol, fumarate, histamine, and histidine. The anomeric region showed signals related to xylose, arabinose, glucose, fructose and sucrose. Finally, in the aliphatic region, signals due to amino acids like alanine, aspartate, asparagine, isoleucine and threonine, organic acids like acetate, lactate, malate, pyruvate, quinate, succinate, and alcohols like 2,3 butanediol, ethanol, and glycerol and sterol were detected. Principal component analysis performed considering NMR data from either all spectral regions and only aromatic region revealed the potentiality to discriminate the yeast action. Signals due to glucose, fructose, glycerol, malate, fumarate, tyrosol, histidine, and histamine resulted discriminant for sample differentiation. This study, the first one by NMR, provides preliminary insights on the metabolic differences among yeast species involved in cider production

Cell Suspensions of Cannabis sativa (var. Futura): Effect of Elicitation on Metabolite Content and Antioxidant Activity

Cannabis sativa L. is one of the most-studied species for its phytochemistry due to the abundance of secondary metabolites, including cannabinoids, terpenes and phenolic compounds. In the last decade, fiber-type hemp varieties have received interest for the production of many specialized secondary metabolites derived from the phenylpropanoid pathway. The interest in these molecules is due to their antioxidant activity. Since secondary metabolite synthesis occurs at a very low level in plants, the aim of this study was to develop a strategy to increase the production of such compounds and to elucidate the biochemical pathways involved. Therefore, cell suspensions of industrial hemp (C. sativa L. var. Futura) were produced, and an advantageous elicitation strategy (methyl jasmonate, MeJA) in combination with precursor feeding (tyrosine, Tyr) was developed. The activity and expression of phenylalanine ammonia-lyase (PAL) and tyrosine aminotransferase (TAT) increased upon treatment. Through 1H-NMR analyses, some aromatic compounds were identified, including, for the first time, 4-hydroxyphenylpyruvate (4-HPP) in addition to tyrosol. The 4-day MeJA+Tyr elicited samples showed a 51% increase in the in vitro assay (2,2-diphenyl-1-picrylhydrazyl, DPPH) radical scavenging activity relative to the control and a 80% increase in the cellular antioxidant activity estimated on an ex vivo model of human erythrocytes. Our results outline the active metabolic pathways and the antioxidant properties of hemp cell extracts under the effect of specific elicitors

New Insight into Justicidin B Pathway and Production in <i>Linum austriacum</i>

Lignans are the main secondary metabolites synthetized by Linum species as plant defense compounds but they are also valuable for human health, in particular, for novel therapeutics. In this work, Linum austriacum in vitro cultures, cells (Cc), adventitious roots (ARc) and hairy roots (HRc) were developed for the production of justicidin B through elicitation with methyl jasmonate (MeJA) and coronatine (COR). The performances of the cultures were evaluated for their stability, total phenols content and antioxidant ability. NMR was used to identify justicidin B and isojusticidin B and HPLC to quantify the production, highlighting ARc and HRc as the highest productive tissues. MeJA and COR treatments induced the synthesis of justicidin B more than three times and the synthesis of other compounds. RNA-sequencing and a de novo assembly of L. austriacum ARc transcriptome was generated to identify the genes activated by MeJA. Furthermore, for the first time, the intracellular localization of justicidin B in ARc was investigated through microscopic analysis. Then, HRc was chosen for small-scale production in a bioreactor. Altogether, our results improve knowledge on justicidin B pathway and cellular localization in L. austriacum for future scale-up processes