IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Unravelling Saccharomyces cerevisiae biosynthethic pathways of melatonin, serotonin and hydroxytyrosol  by UPLC-HRMS Isotopic labelling analysis

Unravelling Saccharomyces cerevisiae biosynthethic pathways of melatonin, serotonin and hydroxytyrosol  by UPLC-HRMS Isotopic labelling analysis

Abstract

The main objective is to unravel the yeast biosynthetic pathways for MEL, SER and HT by using the respective labelled amino acids precursors: 15N2-L tryptophan and 13C-tyrosine.
The alcoholic fermentation experiments are performed with two different commercial S cereviseae yeasts using synthetic must with the addition of the labelled compounds and the bioactive compounds were followed during the fermentation process. Six biological replicates of the fermentations were considered. MEL, SER and HT were analysed by UHPLC coupled to High Resolution Mass Spectrometry (HRMS). Accurate mass determination allowed to unequivocally distinguishing labelled and unlabelled compounds. The analytical determination was performed using external calibration curves with 10 points, which were freshly prepared at every analytical session. Potential intermediates in the synthesis of MEL are detected as labelled metabolites following a time sequence that fits with the pathway described for the synthesis of MEL in mammals. At day 3 all the initial 15N2 L-TRP has been consumed. However, L-TRP as such was detected despite it was not added to the fermentation medium, thus demonstrating de novo tryptophan formation which can be used to synthetize MEL thereafter. Similarly, the results obtained in the fermentation carried out with 13-C TYR verify that HT is formed both from labelled tyrosine and from intermediates of the Erlich pathway.

DOI:

Publication date: June 24, 2022

Issue: IVAS 2022

Type: Poster

Authors

Troncoso Ana M.1, Gallardo-Fernández Marta1, Valls-Fonayet Josep2, Valero Eva3, Hornedo-Ortega Ruth1, Richard Tristan2 and García-Parrilla M Carmen1 

1Universidad de Sevilla
2Université de Bordeaux
3Universidad Pablo de Olavide

Contact the author

Keywords

yeast, alcoholic fermentation, bioactive compounds, isotopically labelled compounds

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Rapid damage assessment and grapevine recovery after fire

There is increasing scientific consensus that climate changeis the underlying cause of the prolonged dry and hot conditions that have increased the risk of extreme fire weather in many countries around the world. In December 2019, a bushfire event occurred in the Adelaide Hills, South Australia where 25,000 hectares were burnt and in vineyards and surrounding areas various degrees of scorching and infrastructure damage occurred. The ability to coordinate and plan recovery after a fire event relies on robust and timely data. The current practice for measuring the scale and distribution of fire damage is to walk or drive the vineyard and score individual vines based on visual observation. The process is time consuming, subjective, or semi-quantitative at best. After the December 2019 fires, it took many months to access properties and estimate the area of vineyard damaged. This study compares the rapid assessment and mapping of fire damage using high-resolution satellite imagery with more traditional ground based measures. Satellite imagery tracking vineyard recovery in the season following the bushfire is being correlated to field assessments of vineyard productivity such as canopy health and development, fertility and carbohydrate storage. Canopy health in the seasons following the fires correlated to the severity of the initial fire damage. Severely damaged vines had reduced canopy growth, were infertile or had very low fertility as well as lower carbohydrate levels in buds and canes during dormancy, which reduced productivity in the seasons following the bushfire event. In contrast, vines that received minor damage were able to recover within 1-2 years. Tools that rapidly and affordably capture the extent and severity of damage over large vineyard area will allow producers, government and industry bodies to manage decisions in relation to fire recovery planning, coordination and delivery, improving the efficiency and effectiveness of their response.

A multidisciplinary approach to evaluate the effects of the training system on the performance of “Aglianico del Vulture” vineyards

Vineyards are complex agro-ecosystems with high spatial and temporal variability. An efficient training system may counteract the adverse effects of this variability. Moreover, considering the climate change issues, choosing an efficient training system that enhances water use and protects the vines from radiative thermal stress has become a priority for the farmers. A multidisciplinary approach that assesses the soil-crop-yield-wine relationships of vineyards in a distributed and holistic way could bring added knowledge on the behavior of the different training systems. This ongoing research aimed to implement a multidisciplinary approach to study the behavior of “Aglianico del Vulture” grapevines trained with two different systems: a spurred cordon (SC) and an “Alberello in parete” (AL), grown in a high-quality wine production area of Basilicata region (Italy). The approach merged several methods and scales of soil, ecophysiology, must/wine quality, and spectral data collection to assess the influence of the training system. Homogeneous zones (HZs) in both training systems were defined through a procedure based on geomorphological classification, unmanned aerial vehicles (UAV) images analysis, and a traditional soil survey supported by geophysical scanning. During the 2021 season, TDR probes monitored soil water content, while grapevine health status was assessed using eco-physiological measurements (LWP, chlorophyll content, PSII photosynthetic efficiency, LAI, and point-based field spectroscopy). These grapevine in-vivo measurements validated the spectral vegetation indexes (NDVI, RENDVI, CVI, and TVI) derived from the UAV multispectral imagery, which monitored the grapevine status in a distributed and non-invasive way. Grape yield, quality of berries, must and wine were measured to assess the effects of the training systems. The first experimental year results showed the variability of the vineyards and revealed relationships among soil parameters, crop characteristics, and vegetation indices of the SC and AL training systems. This multidisciplinary study could bring new insights into the vineyard training system’s effects on grape yield and wine quality.

The modification of cultural practices in grapevine cv. Syrah, does it modify the characteristics of the musts?

The work shows the results of a year of experimentation (2020) in a Syrah variety vineyard in La Roda (Castilla-La Mancha, Spain). The trial approach was on a randomized block design with two factors: Irrigation (I) and Pruning (P).
Irrigation schedules were adjusted to apply amounts close to 1,500 m3/ha. With this provision, 2 different irrigation treatments were proposed: I1) Start of irrigation from pea-sized grape to post-harvest (providing at least 20 % of the total amount of irrigation water to be provided post-harvest); I2) Start of irrigation from pea-sized grape to harvest (usual irrigation practice in the study area). Pruning was proposed with two treatments, one at the end of January (P1), which is pruning on a conventional date; and P2) pruning carried out at the beginning of budding. In total, 4 repetitions were designed with 4 elementary plots, each one of them representing one of the proposed treatments (I1P1; I1P2; I2P1; I2P2). In total, 16 plots were worked on and each elementary plot consisted of 30 strains, distributed in 3 lines.
The productive response was evaluated with the yield results of the harvest harvested at 23 ºBrix. The qualitative response was measured in the musts through the indices of technological (acidity, pH and potassium) and phenolic maturity and aromatic compounds in free and glycosylated fractions. The treatments tested had, in general, an effect on the different variables analyzed.

Inhibition of Oenococcus oeni during alcoholic fermentation by a selected Lactiplantibacillus plantarum strain

The use of selected cultures of the species Lactiplantibacillus plantarum in Oenology has grown in prominence in recent years. While initial applications of this species centred very much around malolactic fermentation (MLF), there is strong evidence to show that certain strains can be harnessed for their bio-protective effects. Unwanted spontaneous MLF during alcoholic fermentation (AF), driven by rogue Oenococcus oeni, is a winemaking deviation that is very difficult to manage when it occurs. This work set out to determine the efficacy of one particular strain of Lactiplantibacillus plantarum(Viniflora® NoVA™ Protect), against this problem in Cabernet Sauvignon must. The work was carried out at commercial scale and in a winery environment and compared the bio-protective culture with the more traditional approach of reducing must pH by the addition of tartaric acid. The combination of both was also investigated. The concentration of both Oenococcus oeni and Lactiplantibacillus plantarum was determined using qPCR. The adventitious Oenococcus oeni showed the most growth during AF in the control wine, whereas in the wines treated with Lactiplantibacillus plantarum a bacteriostatic effect against this species was observed. This effect was comparable to the wines treated with tartaric acid. This has particular commercial relevance for controlling the flora in musts with high pH, or when the addition of tartaric acid is either not permitted or is prohibitive for other reasons.

How can historical cultivars mitigate the effects of climate change?

IFV, INRAe and the national network “Partenaires de la Sélection Vigne” representing 37 organizations from the different wine regions, have been working increasingly closely over the last 2 decades towards the preservation of the French varietal patrimony. There are approximately 600 patrimonial varieties according to INRAe and SupAgro Montpellier experts, including ancient cultivars (400) and intravarietal crossbreeds obtained since the 19th century. In the context of a drastic reduction in such varieties from the mid 1980’s in favor of mainstream varieties, it was essential to carry out an inventory of old vines and vineyards. INRAe Vassal collection plays a key role here as it holds the largest diversity available, along with a rich bibliography and herbariums, offering us the opportunity to document and double check the identity of a cultivar, consolidating the expertise of ampelographers. The work is carried out in several stages, from verifying the existence of a variety in a small region, through to rehabilitation. During this session, the authors present the process that leads to the official registration of a variety. After this, IFV selection center takes over to initiate the process of selection and propagation. A specific focus within regions such as the Alps, Champagne and the South-West will provide details of the full procedure. Bia, Bouysselet, Chardonnay rose, Mecle and the aptly named Tardif, are some of the cultivars that have followed this procedure. Furthermore, a recent regulation established by INAO on “varieties of interest for adaptation purposes” might boost uptake by growers. Since 2006, 36 historical cultivars have been registered. Most of these have been neglected in the past due to late maturity, lack of sugar and high titratable acidity at harvest time. Such characteristics are today considered as positive qualities, not only in mitigation of the effects of climate change, but also as an opportunity for restoring diversity…