Thiol Release Research Articles

Precursors consumption preferences and thiol release capacity of the wine yeasts Saccharomyces cerevisiae, Torulaspora delbrueckii, and Lachancea thermotolerans

The aromatic profile of wine determines its overall final quality, and among the volatile molecules that define it, varietal thiols are responsible for shaping the distinctive character of certain wine varieties. In grape must, these thiols are conjugated to amino acids or small peptides in a non-volatile form. During wine fermentation, yeasts play a principal role in expressing these aromatic compounds as they internalise and cleavage these precursors, releasing the corresponding free and aroma-impacting fraction. Here, we investigate the impact of three wine yeasts (Saccharomyces cerevisiae, Torulaspora delbrueckii and Lachancea thermotolerans) on thiol releasing in synthetic grape must fermentations supplemented with different cysteinylated (Cys-4MSP and Cys-3SH) and glutathionylated (GSH-4MSP and GSH-3SH) precursors. We demonstrate higher consumption levels of cysteinylated precursors, and consequently, higher amounts of thiols are released from them compared to glutathionylated ones. We also report a significant impact of yeast inoculated on the final thiols released. Meanwhile T. delkbrueckii exhibits a great 3SHA releasing capacity, L. thermotolerans stands out because of its high 3SH release. We also highlight the synergic effect of the co-inoculation strategy, especially relevant in the case of S. cerevisiae and L. thermotolerans mixed fermentation, that has an outstanding release of 4MSP thiol. Although our results stem from a specific experimental approach that differs from real winemaking situations, these findings reveal the potential of unravelling the specific role of different yeast species, thiol precursors and their interaction, to improve wine production processes in the context of wine aroma enhancement.

Microbial β C-S Lyases: Enzymes with Multifaceted Roles in Flavor Generation.

β C-S lyases (β-CSLs; EC 4.4.1.8) are enzymes catalyzing the dissociation of β carbon-sulfur bonds of cysteine S-conjugates to produce odorant metabolites with a free thiol group. These enzymes are increasingly studied for their role in flavor generation in a variety of food products, whether these processes occur directly in plants, by microbial β-CSLs during fermentation, or in the mouth under the action of the oral microbiota. Microbial β-CSLs react with sulfur aroma precursors present in beverages, vegetables, fruits, or aromatic herbs like hop but also potentially with some precursors formed through Maillard reactions in cooked foods such as meat or coffee. β-CSLs from microorganisms like yeasts and lactic acid bacteria have been studied for their role in the release of polyfunctional thiols in wine and beer during fermentation. In addition, β-CSLs from microorganisms of the human oral cavity were shown to metabolize similar precursors and to produce aroma in the mouth with an impact on retro-olfaction. This review summarizes the current knowledge on β-CSLs involved in flavor generation with a focus on enzymes from microbial species present either in the fermentative processes or in the oral cavity. This paper highlights the importance of this enzyme family in the food continuum, from production to consumption, and offers new perspectives concerning the utilization of β-CSLs as a flavor enhancer.

Unexpected Behavior of a Maltose-Negative Saccharomyces cerevisiae Yeast: Higher Release of Polyfunctional Thiols from Glutathionylated Than from Cysteinylated S-Conjugates

At present, non-alcoholic and low-alcoholic beers (NABLABs), in addition to their premature sensitivity to oxidation, still suffer from a lack of fruity fermentation aromas. Maltose/maltotriose-negative yeasts offer a highly attractive alternative for creating diversified pleasant aromas and/or eliminating off-flavors in NABLAB production. The aim of this study was to explore the potential of Saccharomyces cerevisiae var. chevalieri, SafBrewTM LA-01 to release fruity polyfunctional thiols from glutathionylated (G-) and cysteinylated (Cys-) precursors. Interestingly, it proved to release free thiols from their glutathionylated S-conjugate much more efficiently (0.34% from G-3-sulfanylhexanol in 15 °P wort after seven days at 24 °C) than the best S. pastorianus strains previously screened (0.13% for lager yeast L7). On the other hand, despite its classification as a S. cerevisiae strain, it showed an inefficient use of cysteinylated precursors, although the release efficiency was slightly higher under NABLAB fermentation conditions (6 °P; 3 days at 20 °C). Under these conditions, as expected, LA-01 consumed only glucose, fructose, and saccharose (0.4% v/v ethanol formation) and produced only low levels of fermentation esters (1.6 mg/L in total) and dimethylsulfide (5 µg/L). The POF+ character of LA-01 also brought significant levels of 4-vinylguaiacol (810 μg/L), which could give to NABLABs the flavors of a white beer.

A novel bioresponsive self-immolative spacer based on aza-quinone methide reactivity for the controlled release of thiols, phenols, amines, sulfonamides or amides.

A stimuli-sensitive linker is one of the indispensable components of prodrugs for cancer therapy as it covalently binds the drug and releases it upon external stimulation at the tumour site. Quinone methide elimination has been widely used as the key transformation to release drugs based on their nucleofugacity. The usual approach is to bind the drug to the linker as a carbamate and release it as a free amine after a self-immolative 1,6-elimination. Although this approach is very efficient, it is limited to amines (as carbamates), alcohols or phenols (as carbonates) or other acidic functional groups. We report here a self-immolative spacer capable of directly linking and releasing amines, phenols, thiols, sulfonamides and carboxyamides after a reductive stimulus. The spacer is based on the structure of (5-nitro-2-pyrrolyl)methanol (NPYM-OH), which was used for the direct alkylation of the functional groups mentioned above. The spacer is metabolically stable and has three indispensable sites for bioconjugation: the bioresponsive trigger, the conjugated 1,6 self-immolative system and a third arm suitable for conjugation with a carrier or other modifiers. Release was achieved by selective reduction of the nitro group over Fe/Pd nanoparticles (NPs) in a micellar aqueous environment (H2O/TPGS-750-M), or by NADH mediated nitroreductase activation. A DFT study demonstrates that, during the 1,6 elimination, the transition state formed from 5-aminopyrrole has a lower activation energy compared to other 5-membered heterocycles or p-aminobenzyl derivatives. The NPYM scaffold was validated by late-stage functionalisation of approved drugs such as celecoxib, colchicine, vorinostat or ciprofloxacin. A hypoxia-activated NPYM-based prodrug (HAP) derived from HDAC inhibitor ST7612AA1 was also produced, which was active in cancer cells under hypoxic conditions.

Carbene-Catalyzed Access to Thiochromene Derivatives: Control of Reaction Pathways via Slow Release of Thiols from Disulfides.

Substrates containing disulfide bonds, which are more stable and less smelling, could be used as thiophenol precursors in organic synthesis. Herein, an N-heterocyclic carbene (NHC)-catalyzed reaction between α-bromoenals and 2,2'-dithiodibenzaldehydes was developed. Through the sustained release strategy, the side reaction can be effectively inhibited, and the chiral thiochromene derivatives can be obtained with good yields and high optical purities. Application studies showed encouraging results when the desired products were explored for antimicrobial utilities in pesticide development.

Unravelling copper effect on the production of varietal thiols during Colombard and Gros Manseng grape juices fermentation by Saccharomyces cerevisiae.

Nowadays the rapidly increasing organic vineyard management with the utilization of copper as sole fungal control pesticide against downy mildew raises once again the question of copper impact on varietal thiols in wine. For this purpose, Colombard and Gros Manseng grape juices were fermented under different copper levels (from 0.2 to 3.88 mg/l) to mimic the consequences in must of organic practices. The consumption of thiol precursors and the release of varietal thiols (both free and oxidized forms of 3-sulfanylhexanol and 3-sulfanylhexyl acetate) were monitored by LC-MS/MS. It was found that the highest copper level (3.6 and 3.88 mg/l for Colombard and Gros Manseng respectively) significantly increased yeast consumption of precursors (by 9.0 and 7.6% for Colombard and Gros Manseng respectively). For both grape varieties, free thiol content in wine significantly decreased (by 84 and 47% for Colombard and Gros Manseng respectively) with the increase of copper in the starting must as already described in the literature. However, the total thiol content produced throughout fermentation was constant regardless of copper conditions for the Colombard must, meaning that the effect of copper was only oxidative for this variety. Meanwhile, in Gros Manseng fermentation, the total thiol content increased along with copper content, resulting in an increase up to 90%; this suggests that copper may modify the regulation of the production pathways of varietal thiols, also underlining the key role of oxidation. These results complement our knowledge on copper effect during thiol-oriented fermentation and the importance of considering the total thiol production (reduced+oxidized) to better understand the effect of studied parameters and differenciate chemical from biological effects.

Examining How the Fermentation Medium Influences Thiol Expression and Its Perceived Aroma in Commercial Brewing Yeast Strains.

In Saccharomyces, the IRC7 gene encodes for a cysteine S-conjugate β-lyase enzyme which can release polyfunctional thiols from their cysteinylated precursor forms, thereby promoting thiol aroma in beer. This study examined the thiol production of 10 commercial yeast strains in two different media, a hopped yeast extract-peptone-dextrose (YPD) medium and a 100% barley malt wort to explore how differences in yeast strain and medium conditions influence the release of polyfunctional thiols. 3-Sulfanylhexan-1-ol was most affected by medium conditions, and its concentrations were highest in wort fermentations. The higher nitrogen content and pH of the YPD medium relative to the wort fermentations were notable differences, and significant correlations between these variables and the extent of free thiol production were observed. A strong association existed between polyfunctional thiol concentrations and the fermentation-derived, malt, and hop-derived compounds 2-phenylethanol, β-damascenone, and β-ionone. The sensory impressions of thiol character in beer were influenced by the presence of other aromatic compounds such as esters and terpene alcohols, and aroma attributes such as "tropical" were not the most suitable for describing beers brewed with yeasts that fully express homozygous IRC7F. Sensory attributes "sweaty", "vegetal", and "overripe fruit" were more strongly associated with these strains.

Hydrogen sulfide production during early yeast fermentation correlates with volatile sulfur compound biogenesis but not thiol release.

Yeasts undergo intensive metabolic changes during the early stages of fermentation. Previous reports suggest the early production of hydrogen sulfide (H2S) is associated with the release of a range of volatile sulfur compounds (VSCs), as well as the production of varietal thiol compounds 3-sulfanylhexan-1-ol (3SH) and 3-sulfanylhexyl acetate (3SHA) from six-carbon precursors including (E)-hex-2-enal. In this study, we investigated the early H2S potential, VSCs/thiol output, and precursor metabolism of 11 commonly used laboratory and commercial Saccharomyces cerevisiae strains in chemically defined synthetic grape medium (SGM) within 12 hours after inoculation. Considerable variability in early H2S potential was observed among the strains surveyed. Chemical profiling suggested that early H2S production correlates with the production of dimethyl disulfide, 2-mercaptoethanol, and diethyl sulfide, but not with 3SH or 3SHA. All strains were capable of metabolizing (E)-hex-2-enal while the F15 strain showed significantly higher residue at 12 h. Early production of 3SH, but not 3SHA, can be detected in the presence of exogenous (E)-hex-2-enal and H2S. Therefore, the natural variability of early yeast H2S production contributes to the early output of selected VSCs, but the threshold of which is likely not high enough to contribute substantially to free varietal thiols in SGM.

High power ultrasound treatment of crushed grapes: Beyond the extraction phenomena

The treatment of white and red crushed grapes by high power ultrasounds (US) represents an emerging technology in winemaking. In 2019, it was officially recognized by OIV through the resolution n°616-2019, and it was also approved by European Union in January 2022. The US effect on extraction mechanisms was widely studied, but more researches are needed to better understand the ultrasound effect on some specific classes of grape compounds. This research aimed to highlight at laboratory scale some specific effects of ultrasounds on some key compounds of white and red grapes. The samples were sonicated at different frequency (20-30 kHz), time (1-10 min), and power (30-90%) technological conditions used in maceration, to obtain valuable information on potential technological transferability. Valuable results were obtained regarding the release of thiols from their precursors, and the reactivity changes of unstable proteins of white wines. The experimental trails on red grape varieties allowed a maintenance of free anthocyanins and no degradative effects were highlighted. Significant and valuable effects were determined also on the tannin polymerization, with an astringency decrease. The sonication treatment of crushed grapes showed several chemical effects that contribute to decreasing the winemaking inputs and preserving the wine quality. The process conditions must be managed related to grape variety and ripeness for a precision winemaking.

A unique reaction of diphenylcyclopropenone and 1,2-aminothiol with the release of thiol for multiple bioconjugation.

Selective reaction of diphenylcyclopropenone (DPCP) and 1,2-aminothiol in water at pH 7.4 produces an amide conjugate with the release of thiol. In addition, structural modifications of DPCP enable the coupling rate to be tuned with a reaction constant of +3.68. Based on this chemistry, triple labelling was demonstrated by treating an N-terminal cysteine peptide with DPCP-Cl followed by thiol-maleimide and tyrosine-diazonium couplings in one pot. We anticipate that the DPCP motif will be a useful toolkit for multiple bioconjugation.

Increased volatile thiol release during beer fermentation using constructed interspecies yeast hybrids

Interspecies hybridization has been shown to be a powerful tool for developing and improving brewing yeast in a number of industry-relevant respects. Thanks to the popularity of heavily hopped ‘India Pale Ale’-style beers, there is an increased demand from brewers for strains that can boost hop aroma. Here, we explored whether hybridization could be used to construct strains with an enhanced ability to release hop-derived flavours through β-lyase activity, which releases desirable volatile thiols. Wild Saccharomyces strains were shown to possess high β-lyase activity compared to brewing strains, however, they also produced phenolic off-flavours (POF) and showed poor attenuation. To overcome these limitations, interspecies hybrids were constructed by crossing pairs of one of three brewing and one of three wild Saccharomyces strains (S. uvarum and S. eubayanus). Hybrids were screened for fermentation ability and β-lyase activity, and selected hybrids showed improved fermentation and formation of both volatile thiols (4MMP, 3MH and 3MH-acetate) and aroma-active esters compared to the parent strains. Undesirable traits (e.g. POF) could be removed from the hybrid by sporulation. To conclude, it was possible to boost the release of desirable hop-derived thiols in brewing yeast by hybridization with wild yeast. This allows production of beer with boosted hop aroma with less hops (thus improving sustainability issues).

Presence of Disulfide-Bonded Thiols in Malt and Hops as the Precursors of Thiols in Beer.

Disulfide-bonded thiols in malt and hops were first identified as possible precursors of thiols in beer. The presence of disulfide-bonded 3-mercaptohexan-1-ol (3MH) was confirmed in malt and hops by observing an 8.9-9.9 times increase in the 3MH concentration in hopped water and unhopped wort after the reduction using tris(2-carboxyethyl)phosphine (TCEP), a reducing agent specific for disulfide bonds. The presence of disulfide-bonded 4-mercapto-4-methylpentan-2-one (4MMP) was confirmed in hops by observing 2.1 and 5.1 times increase in the 4MMP concentration after reduction in hopped water. Proteins, peptides, and amino acids having sulfhydryl groups or other thiol substances were assumed to form disulfide bonds with polyfunctional thiols in malt and hops. The release of thiols by the reduction of disulfide-bonded thiols during fermentation was first identified. A 65-82% of disulfide-bonded 3MH were reduced during fermentation, and as a result, concentrations of 3MH in hopped water and unhopped wort increased by 9.5-14.2 times during fermentation.

Modulation of Volatile Thiol Release during Fermentation of Red Musts by Wine Yeast

During the alcoholic fermentation of grape sugars, wine yeast produces a range of secondary metabolites that play a critical role in the aroma profile of wines. One of the most impactful yeast-modified compound families, particularly in white wines, are the ‘fruity’ polyfunctional thiols, which include 3-mercaptohexan-1-ol (3-MH) and 4-mercapto-4-methylpentan-2-one (4-MMP). While the formation and stylistic contribution of these thiols have been extensively researched in white wines, little is known about the conditions leading to their formation in red wines. In this study, we explored the ability of yeast strains to modulate the release of these aroma compounds during the fermentation of two red musts. In laboratory-scale Pinot Noir fermentations, the formation of 3-MH strongly correlated with yeast β-lyase activity, particularly with the presence of certain genotypes of the flavour-releasing gene IRC7. Subsequent production of Grenache wine at the pilot scale, with detailed compositional and sensory analysis, was undertaken to confirm laboratory-scale observations. A commercial wine strain used for expressing ‘fruity’ thiols in Sauvignon Blanc was shown to produce wines that exhibited more intense red fruit aromas. These results reveal an opportunity for winemakers to shape red wine aroma and flavour by using yeasts that might typically be considered for white wine production.

Effect of Addition of Tryptophan on Aggregation of Apo-α-Lactalbumin Induced by UV-Light.

UV-B illumination facilitates aggregation of alpha-lactalbumin (α-LA) by intramolecular disulfide bond cleavage followed by intermolecular thiol-disulfide exchange reactions. However, long term exposure to UV-B illumination may induce undesired oxidative modifications of amino acid residues in the protein. The purpose of this study was to examine the effect of UV-induced aggregation of apo-α-LA (a calcium-depleted form of α-LA) under aerobic and anaerobic conditions and by addition of tryptophan (Trp) as a photosensitizer. The addition of Trp to apo-α-LA illuminated under anaerobic conditions facilitated the highest level of free thiol release and disulfide-mediated aggregation as compared to without addition of Trp under both anaerobic and aerobic conditions. Addition of Trp under aerobic condition resulted in the lowest level of free thiols and disulfide-mediated aggregation and the aerobic conditions caused oxidation of the free Trp with formation of kynurenine and 5-hydroxy-Trp. Minor levels of the Trp oxidation product, 3-hydroxy-kynurenine (2% converted from Trp), was formed in apo-α-LA with added Trp under both aerobic and anaerobic conditions after UV-B treatment.

Modulation of the Sulfanylalkyl Acetate/Alcohol Ratio and Free Thiol Release from Cysteinylated and/or Glutathionylated Sulfanylalkyl Alcohols in Beer under Different Fermentation Conditions

The occurrence of a substantial pool of cysteinylated and glutathionylated forms of polyfunctional thiols has been evidenced for several dual-purpose hop varieties, and so is the ability of Saccharomyces cerevisiae yeast to release free thiols from these forms through fermentation. The present work aimed to investigate the effect of temperature, wort density, maturation time, and strain on the efficiency of free thiol release by S. cerevisiae yeasts. Model media at 12, 15, or 17°P were spiked with three cysteinylated (Cys-) or three glutathionylated (G-) sulfanylalkyl alcohols (Cys- or G-3-sulfanylpentan-1-ol, 3-sulfanyl-4-methylpentan-1-ol, and 3-sulfanylhexan-1-ol), fermented for 7 days at 18, 24, and 28 °C, and kept at 4 °C for varying number of days. The released sulfanylalkyl alcohols and their corresponding acetates were extracted with a Ag-ion SPE cartridge and analyzed by gas chromatography-pulsed-flame photometric detection. The wort density and yeast strain greatly affected the acetate/alcohol ratio. This ratio varied from 1 to 80% according to the yeast strain and was at its highest at 17°P and 24 °C. Maturation appeared as the crucial step for free thiol excretion from yeast cells (no thiol was recovered in the fermented worts without maturation). Among the five yeasts tested, the yeast strain SafAle K-97 released the highest level of sulfanylalkyl alcohols into the medium (up to 0.45% of the added cysteinylated adducts and 0.08% of the glutathionylated adducts), whereas S-33 or S-04 should be preferred when release of esters is sought out (release efficiencies up to 0.35% from cysteinylated adducts and 0.02% from glutathionylated adducts are observed if both the alcohol and its acetate are considered).

Вміст оксиду азоту та S-нітрозотіолів у клітинах печінки щурів за умов різного забезпечення харчового раціону макронутрієнтами

This paper presents studies of nitric oxide and low-molecular S-nitrosothiols in the mitochondrial and cytosolic fractions of the rats' liver under the conditions of, alimentary protein deprivation, consumption of excess sucrose content and combined action of two adverse factors. In order to model the low-protein diet of the animal for 28 days received an isocaloric diet containing 4.7% protein, 10% fat, 81,3% carbohydrates (starch – 37%, sucrose – 30%, cellulose – 5%) and was calculated in accordance with the recommendations of the American Institute of Nutrition. The high-sugar diet consisted of 14% protein, 10% fat, 72% carbohydrates (starch – 37%, sucrose – 30%, cellulose – 5%). The mitochondrial and cytosolic fraction of rat liver cells were obtained by the method of differential centrifugation. Nitrogen oxide content was assessed by a unified method by determining the NO2- content, which is a stable metabolite of nitric oxide. Since NO is inactivated into an oxidase reaction with the conversion into nitrite or nitrate that is quickly metabolized, the nitrogen oxide content was assessed by the change in NO2-. The concentration of S-nitrosothiols was recorded, respectively, by determining the concentration of nitrite anion before and after the addition of Hg2+ ions, which by modifying the S – N bonds catalyzes the release of S-nitrosyl thiols of nitric oxide. An increase in NO content in both hepatic subcellular fractions of the rats’ experimental groups compared to control values was found. However, a lack of protein in the diet (protein deficiency in the diet leads to an increase in nitric oxide levels in 3-4 times) can be considered as a key factor in the recorded changes in the mitochondria of the animals’ liver, while in the cytosol - excessive consumption of sucrose (3-5 times increase). Regarding the level of S-nitrosothiols, in the studied fractions, multidirectional changes in their concentration were found. Thus, an increase in the content of nitrosyl derivatives in the mitochondria of rat’s liver cells with a simultaneous decrease in their level in the cytosol indicates dysmetabolic disorders in the transport system and deposition of nitric oxide, which can lead to the development of nitrosative stress under the experimental conditions.

Fine-tuning thio-pyridazinediones as SMDC scaffolds (with intracellular thiol release via a novel self-immolative linker).

Herein we report the synthesis of a library of thioalkyl- and thioaryl-pyridazinediones for thiol-based self-immolative release of cargo. A bisthioaryl-pyridazinedione is shown to be stable to serum protein albumin but unstable in intracellular conditions. A derivatised analogue underwent self-immolative degradation in cellular thiol conditions as evidenced by LC-MS/release of a turn-on fluorescence fluorophore; versatility of the thiol-pyridazinedione is demonstrated through synthesis of SMDC precursors that contain three different functional groups on the same central molecule.

Disulfide-Mediated Bioconjugation: Disulfide Formation and Restructuring on the Surface of Nanomanufactured (Microfluidics) Nanoparticles.

This study is about (1) nanomanufacturing (focusing on microfluidic-assisted nanoprecipitation), (2) advanced colloid characterization (focusing on field flow fractionation), and (3) the possible restructuring of surface disulfides. Disulfides are dynamic and exchangeable groups, and here we specifically focus, first, on their use to introduce biofunctional groups and, second, on their re-organization, which may lead to variable surface chemistries and uncontrolled cell interactions. The particles were obtained via microfluidic-assisted (flow-focused) nanoprecipitation of poly(ethylene glycol)-b-poly(ε-caprolactone) bearing or not a 2-pyridyl disulfide (PDS) terminal group, which quantitatively exchanges with thiols in solution. In this study, we have paid specific attention to size characterization, thereby also demonstrating the limitations of dynamic light scattering (DLS) as a stand-alone technique. By using asymmetric flow field flow fractionation coupled with DLS, static light scattering (SLS), and refractive index detectors, we show that relatively small amounts of >100 nm aggregates (cryogenic transmission electron microscopy and SLS/DLS comparison suggesting them to be wormlike micelles) dominated the stand-alone DLS results, whereas the "real" size distributions picked <50 nm. Our key result is that the kinetics of the conjugation based on PDS-thiol exchange was controlled by the thiol pKa, and this also determined the rate of the exchange between the resulting disulfides and glutathione (GSH). In particular, more acidic thiols (e.g., peptides, where a cysteine is flanked by cationic residues) react faster with PDS, but their disulfides hardly exchange with GSH; the reverse applies to thiols with a higher pKa. Disulfides that resist against restructuring via thiol-disulfide exchange allow for a stable bioconjugation, although they may be bad news for payload release under reducing conditions. However, experiments of both thiol release and nanoparticles uptake in cells (HCT116) show that also the disulfides formed from less-acidic and, therefore, less-reactive, and more exchangeable thiols were stable for at least a few hours even in a GSH-rich (10 mM) environment; this suggests a sufficiently long stability of surface groups to achieve, for example, a cell-targeting effect.

The molecular biology of fruity and floral aromas in beer and other alcoholic beverages.

Aroma compounds provide attractiveness and variety to alcoholic beverages. We discuss the molecular biology of a major subset of beer aroma volatiles, fruity and floral compounds, originating from raw materials (malt and hops), or formed by yeast during fermentation. We introduce aroma perception, describe the most aroma-active, fruity and floral compounds in fruits and their presence and origin in beer. They are classified into categories based on their functional groups and biosynthesis pathways: (1) higher alcohols and esters, (2) polyfunctional thiols, (3) lactones and furanones, and (4) terpenoids. Yeast and hops are the main sources of fruity and flowery aroma compounds in beer. For yeast, the focus is on higher alcohols and esters, and particularly the complex regulation of the alcohol acetyl transferase ATF1 gene. We discuss the release of polyfunctional thiols and monoterpenoids from cysteine- and glutathione-S-conjugated compounds and glucosides, respectively, the primary biological functions of the yeast enzymes involved, their mode of action and mechanisms of regulation that control aroma compound production. Furthermore, we discuss biochemistry and genetics of terpenoid production and formation of non-volatile precursors in Humulus lupulus (hops). Insight in these pathways provides a toolbox for creating innovative products with a diversity of pleasant aromas.

Lipids modulate acetic acid and thiol final concentrations in wine during fermentation by Saccharomyces cerevisiae\u2009\xd7\u2009Saccharomyces kudriavzevii hybrids

Saccharomyces cerevisiae × Saccharomyces kudriavzevii hybrids are typically used for white wine fermentation because of their cryotolerance. One group of these hybrids presents a unique ability to release thiol varietal aroma products as well as excessive amounts of acetic acid under specific conditions, which is detrimental for wine organoleptic quality. The aim of this work is to better assess the effects of lipids, sugar concentrations and temperature on the production of acetic acid and thiols during wine fermentation. To this end, we used a Box–Behnken experimental design and response surface modeling on the production of acetic acid and thiols in S. cerevisiae × S. kudriavzevii hybrids from the Eg8 family during fermentation of a synthetic must. We showed that these hybrids produced lower levels of acetic acid when the initial lipid concentration was increased, whereas they produced greater levels when the initial sugar concentration was high. Moreover, we found that lipids had a positive impact on the final concentrations of 4-methyl-4-mercaptopentan-2-one and 3-mercaptohexan-1-ol (3MH), giving box tree and citrus flavors, respectively. The increase of 3MH was concomitant with a decrease of 3-mercaptohexyl acetate (3MHA) characterized by a passion fruit aroma, indicating that lipid addition reduces the rate of 3MH acetylation into 3MHA. These results highlight the key role of lipid management in acetic acid metabolism and thiol release by S. cerevisiae × S. kudriavzevii hybrids and underline its technological interest in alcoholic fermentation to avoid the overproduction of volatile acidity while favoring the release of volatile thiols.