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The desirable wine aroma compounds 3-sulfanylhexan-1-ol (3SH) and 3-sulfanylhexyl acetate (3SHA) are released during fermentation from non-volatile precursors present in the grapes. This work explores the relative contribution of four precursors (E-2-hexenal, 3-S-glutathionylhexan-1-ol, 3-S-glutathionylhexanal, and 3-S-cysteinylhexan-1-ol) to 3SH and 3SHA. Through the use of isotopically labelled analogues of these precursors in defined fermentation media, new insights into the role of each precursor have been identified. E-2-Hexenal was shown to contribute negligible amounts of thiols, while 3-S-glutathionylhexan-1-ol was the main precursor of both 3SH and 3SHA. The glutathionylated precursors were both converted to 3SHA more efficiently than 3-S-cysteinylhexan-1-ol. Interestingly, 3-S-glutathionylhexanal generated 3SHA without detectable concentrations of 3SH, suggesting possible differences in the way this precursor is metabolised compared to 3-S-glutathionylhexan-1-ol and 3-S-cysteinylhexan-1-ol. We also provide the first evidence for chemical conversion of 3-S-glutathionylhexan-1-ol to 3-S-(γ-glutamylcysteinyl)-hexan-1-ol in an oenological system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

3 S -Gluthathionylhexanal (glut3SHal) is an early precursor to the important wine aroma compound 3-sulfanylhexan-1-ol (3SH), imparting tropical passion fruit aromas, even at trace concentrations. In wine, glut3SHal occurs in equilibrium with its bisulfite adduct (glut3SH-SO 3 ), challenging its quantification. To circumvent the issues encountered when attempting to describe the equilibrium between these compounds, a method for their quantification in wine samples was developed. Separation of glut3SHal and glut3SH-SO 3 using solid-phase extraction followed by oxime derivatization and analysis via liquid chromatography-mass spectrometry allowed for measurement of both compounds in wine samples. Analysis of commercial Sauvignon Blanc wines using the developed method confirmed that glut3SH-SO 3 is the major species in the wine matrix. The method developed in this work will enable further exploration of the relationship between glut3SHal and glut3SH-SO 3 and their contribution to production of 3SH in wines. There is potential to extrapolate this work to explore other aldehyde-sulfonic acid equilibria in foods and beverages.

Pinot noir grapes require careful management in the winery to prevent loss of color density and promote aging stability. Winemaking with flocculent yeast has been shown to increase color density, which is desirable to consumers. This research explored interspecies sequential inoculation and co-flocculation of commercial yeast on Pinot noir wine color. Sedimentation rates of six non-Saccharomyces species and two Saccharomyces cerevisiae strains were assayed individually and in combination. The most flocculent pairings, Torulaspora delbrueckii BIODIVA with S. cerevisiae RC212 or VL3, were used to ferment 20 L Pinot noir must. Sequential fermentations produced wines with greater color density at 420 + 520 nm, confirmed by sensory panel. Total and monomeric anthocyanin concentrations were decreased in sequentially fermented wines, despite being the main source of red wine color. BIODIVA adsorbed more anthocyanins than S. cerevisiae, indicating a greater number of cell wall mannoproteins in flocculent yeast, that could then result in a later release of anthocyanins and enhance copigment formation in red wines., (© 2023 The Authors. Yeast published by John Wiley & Sons Ltd.)

Improvements to the quantification of three white wine impact odorants 3-sulfanylhexan-1-ol, 3-sulfanylhexyl acetate, and 4-sulfanyl-4-methylpentan-2-one, and the non-volatile precursors from which they are released during fermentation, is of great interest to the wine science community. Recent reports of a "Quick, Easy, Cheap, Effective, Rugged and Safe" (QuEChERS) based method for the concurrent analysis of these thiols and their precursors via liquid chromatography tandem mass spectrometry (LC-MS/MS) has enabled the development of far simpler methods, as well as aligning these analyses with principles of green analytical chemistry. This current work reports the development and validation of a QuEChERS based LC-MS/MS method utilising a safer derivatising agent, 4,4'-dithiodipyridine, while greatly minimising the reagents involved and waste produced. We demonstrate that this new method compares favourably to the previously reported method with repeatability of 0.2-1.3%RSD and 0.4-5.2%RSD for precursors and free thiols. Further, the commercially available internal standard, 1-hexanethiol, used in previous analytical methods was compared to stable isotope labelled analogues of the analytes, with results suggesting that it may not be a reliable internal standard., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

γ-Nonalactone is a linear aliphatic lactone ubiquitous in wine, associated with coconut, sweet, and stone fruit aroma descriptors. Little research has been conducted looking at the importance of this compound to New Zealand (NZ) wine aroma. 2 H 2 13 C 2 -γ-Nonalactone, a novel isotopologue of γ-nonalactone, was synthesised in this work for use in a stable isotope dilution assay (SIDA) for quantification of γ-nonalactone in NZ Pinot noir wines for the first time. Synthesis was carried out using heptaldehyde as the starting material, and 13 C atoms and 2 H atoms were introduced via Wittig olefination and deuterogenation steps, respectively. The suitability of this compound as an internal standard was demonstrated by spiking model wine at normal and elevated conditions during sample preparation, with subsequent analysis via mass spectrometry showing stability of 2 H 2 13 C 2 -γ-nonalactone. A model wine calibration, with concentrations of γ-nonalactone from 0 to 100 µg L -1 , was shown to have excellent linearity (R 2  > 0.99), reproducibility (0.72%), and repeatability (0.38%). Twelve NZ Pinot noir wines, representative of a range of NZ Pinot noir-producing regions, prices, and vintages, were analysed by solid-phase extraction-gas chromatography-mass spectrometry (SPE-GC-MS). The concentrations of γ-nonalactone ranged from 8.3 to 22.5 µg L -1 , the latter of which was close to the odour detection threshold of this compound. These findings provide a basis for further research into γ-nonalactone and its impact on NZ Pinot noir aroma and provide a robust method for the quantification of this compound in Pinot noir., (© 2023. The Author(s).)

Sensory attributes are essential factors in determining the quality of wines. However, it can be challenging for consumers, even experts, to differentiate and quantify wines' sensory attributes for quality control. Soft sensors based on rapid chemical analysis offer a potential solution to overcome this challenge. However, the current limitation in developing soft sensors for wines is the need for a significant number of input parameters, at least 12, necessitating costly and time-consuming analyses. While such a comprehensive approach provides high accuracy in sensory quality mapping, the expensive and time-consuming studies required do not lend themselves to the industry's routine quality control activities. In this work, Box plots, Tucker-1 plots, and Principal Component Analysis (PCA) score plots were used to deal with output data (sensory attributes) to improve the model quality. More importantly, this work has identified that the number of analyses required to fully quantify by regression models and qualify by classification models can be significantly reduced. Based on regression models, only four key chemical parameters (total flavanols, total tannins, A 520nm HCl , and pH) were required to accurately predict 35 sensory attributes of a wine with R 2 values above 0.6 simultaneously. In addition, for classification models to accurately predict 35 sensory attributes of a wine at once with prediction accuracy above 70%, only four key chemical parameters (A 280nm HCl , A 520nm HCl , chemical age and pH) were required. These models with reduced chemical parameters complement each other in sensory quality mapping and provide acceptable accuracy. The application of the soft sensor based on these reduced sets of key chemical parameters translated to a potential reduction in analytical cost and labour cost of 56% for the regression model and 83% for the classification model, respectively, making these models suitable for routine quality control use., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors. Published by Elsevier B.V.)

Polysulfide degradation in wine can result in hydrogen sulfide (H 2 S) release, imparting a rotten-egg smell that is detrimental to wine quality. Although the presence of wine polysulfides has been demonstrated, their biogenesis remains unclear. This study investigated the role of Saccharomyces cerevisiae in polysulfide formation during fermentation, with and without 5 mM cysteine supplementation as an H 2 S source. Using an established liquid chromatography-tandem mass spectrometry method, monobromobimane derivatives of hydropolysulfides, including CysSSSH, CysSSSSH and GSSSSH, and two oxidized polysulfides, GSSG and GSSSSG, were detected in yeast cells at the end of fermentation in a grape juice-like medium. Polysulfide production by four S. cerevisiae single deletion mutants (BY4743 Δcys3, Δcys4, Δmet17 and Δtum1) showed no significant differences compared to BY4743, suggesting that uncharacterized pathways maintain cellular polysulfide homeostasis. Five mM cysteine addition increased the formation of shorter sulfur chain species, including GSS-bimane and GSSG, but did not elevate levels of longer sulfur chain species. Additionally, polysulfides with even numbers of sulfur atoms tended to predominate in cellular lysates. Oxidized polysulfides and longer chain hydropolysulfides were not detected in finished wines. This evidence suggests that these polysulfides are unstable in wine-like environments or not transported extracellularly. Collectively, our data illustrate the complexity of yeast polysulfide metabolism under fermentation conditions., Competing Interests: Declaration of competing interest Declaration of interest: none., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Soft sensors work as predictive frameworks encapsulating a set of easy-to-collect input data and a machine learning method (ML) to predict highly related variables that are difficult to measure. The machine learning method could provide a prediction of complex unknown relations between the input data and desired output parameters. Recently, soft sensors have been applicable in predicting the prices and vintages of New Zealand Pinot noir wines based on chemical parameters. However, the previous sample size did not adequately represent the diversity of provenances, vintages, and price points across commercially available New Zealand Pinot noir wines. Consequently, a representative sample of 39 commercially available New Zealand Pinot noir wines from diverse provenances, vintages, and price points were selected. Literature has shown that wine phenolic compounds strongly correlated with wine provenances, vintages and price points, which could be used as input data for developing soft sensors. Due to the significance of these phenolic compounds, chemical parameters, including phenolic compounds and pH, were collected using UV-Vis visible spectrophotometry and a pH meter. The soft sensor utilising Naive Bayes (belongs to ML) was designed to predict Pinot noir wines' provenances (regions of origin) based on six chemical parameters with the prediction accuracy of over 75%. Soft sensors based on decision trees (within ML) could predict Pinot noir wines' vintages and price points with prediction accuracies of over 75% based on six chemical parameters. These predictions were based on the same collected six chemical parameters as aforementioned.

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 h 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., (© The Author(s) 2023. Published by Oxford University Press on behalf of FEMS.)

Saturated linear aliphatic lactones are widespread aroma compounds in wine, linked to stone fruit, dried red fruit, and coconut descriptors. Despite their ubiquity, bioproduction pathways associated with these compounds in wine are unclear, but higher concentrations have been linked to many common vitivinicultural practices, including grape variety, microbiological influence, oak- and bottle-aging, and wine styles such as late harvest, noble rot, and icewine. Development of analytical techniques has enabled increasingly accurate quantification of lactones in wine, shedding more light on their potential origins. This review provides an in-depth summary of the research into linear aliphatic lactones over the past 50 years and provides direction for possible future research to elucidate the biogenesis of these compounds and better estimate their impact on wine aroma.

Volatile polyfunctional thiol compounds, particularly 3-sulfanylhexan-1-ol (3SH) and 3-sulfanylhexyl acetate (3SHA), are key odorants contributing to the aroma profile of many wine styles, generally imparting tropical grapefruit and passionfruit aromas. 3SH and 3SHA are present in negligible concentrations in the grape berry, juice, and must, suggesting that they are released from non-volatile precursors present in the grape. The exploration of the nature and biogenesis of these precursors to 3SH and 3SHA has proven important for the elucidation of polyfunctional thiol biogenesis during alcoholic fermentation. The development and validation of appropriate analytical techniques for the analysis of 3SH precursors in enological matrices have been extensive, and this review explores the analysis and discovery of these precursor compounds. The development of analytical methods to analyze 3SH precursors, from the selection of the analytical instrument, sample preparation, and methods for standardization, will first be discussed, before highlighting how these techniques have been used in the elucidation of the biogenesis of 3SH and 3SHA in grape wines. Lastly, the future of thiol precursor analysis will be considered, with the development of new methods that greatly reduce the sample preparation time and enable multiple precursors, and the thiols themselves, to be quantitated using a single method.

As part of our search for new antimicrobials and antibiotic adjuvants, a series of podocarpic acid-polyamine conjugates have been synthesized. The library of compounds made use of the phenolic and carboxylic acid moieties of the diterpene allowing attachment of polyamines (PA) of different lengths to afford a structurally-diverse set of analogues. Evaluation of the conjugates for intrinsic antimicrobial properties identified two derivatives of interest: a PA3-4-3 (spermine) amide-bonded variant 7a that was a non-cytotoxic, non-hemolytic potent growth inhibitor of Gram-positive Staphylococcus aureus (MRSA) and 9d, a PA3-8-3 carbamate derivative that was a non-toxic selective antifungal towards Cryptococcus neoformans. Of the compound set, only one example exhibited activity towards Gram-negative bacteria. However, in the presence of sub-therapeutic amounts of either doxycycline (4.5 µM) or erythromycin (2.7 μM) several analogues were observed to exhibit weak to modest antibiotic adjuvant properties against Pseudomonas aeruginosa and/or Escherichia coli. The observation of strong cytotoxicity and/or hemolytic properties for subsets of the library, in particular those analogues bearing methyl ester or n-pentylamide functionality, highlighted the fine balance of structural requirements and lipophilicity for antimicrobial activity as opposed to mammalian cell toxicity., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The volatile thiol compound 3-sulfanylhexan-1-ol (3SH) is a key impact odorant of white wines such as Sauvignon Blanc. 3SH is produced during fermentation by metabolism of non-volatile precursors such as 3- S -gluthathionylhexanal (glut-3SH-al). The biogenesis of 3SH is not fully understood, and the role of glut-3SH-al in this pathway is yet to be elucidated. The aldehyde functional group of glut-3SH-al is known to make this compound more reactive than other precursors to 3SH, and we are reporting for the first time that glut-3SH-al can exist in both keto and enol forms in aqueous solutions. At wine typical pH (~3.5), glut-3SH-al exists predominantly as the enol form. The dominance of the enol form over the keto form has implications in terms of potential consumption/conversion of glut-3SH-al by previously unidentified pathways. Therefore, this work will aid in the further elucidation of the role of glut-3SH-al towards 3SH formation in wine, with significant implications for the study and analysis of analogous compounds.

It is standard practice to ferment white wines at low temperatures (10-18°C). However, low temperatures increase fermentation duration and risk of problem ferments, leading to significant costs. The lag duration at fermentation initiation is heavily impacted by temperature; therefore, identification of Saccharomyces cerevisiae genes influencing fermentation kinetics is of interest for winemaking. We selected 28 S. cerevisiae BY4743 single deletants, from a prior list of open reading frames (ORFs) mapped to quantitative trait loci (QTLs) on Chr. VII and XIII, influencing the duration of fermentative lag time. Five BY4743 deletants, Δapt1, Δcgi121, Δclb6, Δrps17a, and Δvma21, differed significantly in their fermentative lag duration compared to BY4743 in synthetic grape must (SGM) at 15 °C, over 72 h. Fermentation at 12.5°C for 528 h confirmed the longer lag times of BY4743 Δcgi121, Δrps17a, and Δvma21. These three candidates ORFs were deleted in S. cerevisiae RM11-1a and S288C to perform single reciprocal hemizygosity analysis (RHA). RHA hybrids and single deletants of RM11-1a and S288C were fermented at 12.5°C in SGM and lag time measurements confirmed that the S288C allele of CGI121 on Chr. XIII, encoding a component of the EKC/KEOPS complex, increased fermentative lag phase duration. Nucleotide sequences of RM11-1a and S288C CGI121 alleles differed by only one synonymous nucleotide, suggesting that intron splicing, codon bias, or positional effects might be responsible for the impact on lag phase duration. This research demonstrates a new role of CGI121 and highlights the applicability of QTL analysis for investigating complex phenotypic traits in yeast., (© The Author(s) 2021. Published by Oxford University Press on behalf of Genetics Society of America.)

Saccharomyces cerevisiae yeasts are a diverse group of single-celled eukaryotes with tremendous phenotypic variation in fermentation efficiency, particularly at different temperatures. Yeast can be categorized into subsets based on lifestyle (Clinical, Fermentation, Laboratory, and Wild), genetic lineage (Malaysian, Mosaic, North American, Sake, West African, and Wine), and geographical origin (Africa, Americas, Asia, Europe, and Oceania) to start to understand their ecology; however, little is known regarding the extent to which these groupings drive S. cerevisiae fermentative ability in grape juice at different fermentation temperatures. To investigate the response of yeast within the different subsets, we quantified fermentation performance in grape juice by measuring the lag time, maximal fermentation rate ( V max ), and fermentation finishing efficiency of 34 genetically diverse S. cerevisiae strains in grape juice at five environmentally and industrially relevant temperatures (10, 15, 20, 25, and 30 °C). Extensive multivariate analysis was applied to determine the effects of lifestyle, lineage, geographical origin, strain, and temperature on yeast fermentation phenotypes. We show that fermentation capability is inherent to S. cerevisiae and that all factors are important in shaping strain fermentative ability, with temperature having the greatest impact, and geographical origin playing a lesser role than lifestyle or genetic lineage.

Volatile sulfur compounds (VSCs) greatly influence the sensory properties and quality of wine and arise via both biological and chemical mechanisms. VSCs formed can also act as precursors for further downstream VSCs, thus elucidating the pathways leading to their formation is paramount. Short-term additions of exogenous hydrogen sulfide (H 2 S), ethanethiol (EtSH), S-ethylthio acetate (ETA), methanethiol (MeSH) and S-methylthio acetate (MTA) were made to exponentially growing fermentations of synthetic grape medium. The VSC profiles produced from live yeast cells were compared with those from dead cells and no cells. Interestingly, this experiment allowed the identification of specific biochemical and/or chemical pathways; e.g. most of the conversion of H 2 S to EtSH, and the further step from EtSH to ETA, required the presence of live yeast cells, as did the conversion of MeSH to MTA. In contrast, the reaction from MTA to MeSH and ETA to EtSH was due primarily to chemical degradation. Ultimately, this research unravelled some of the complex interactions and interconversions between VSCs, pinpointing the key biochemical and chemical nodes. These pathways are highly interconnected and showcase the complexity of both the sulfur pathways in yeast and the reactive chemistry of sulfur-containing compounds., Competing Interests: Declaration of competing interest The authors declares no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In an effort to gain more understanding on the structure activity relationship of pseudoceratidine 1 , a di-bromo pyrrole spermidine alkaloid derived from the marine sponge Pseudoceratina purpurea that has been shown to exhibit potent biofouling, anti-fungal, antibacterial, and anti-malarial activities, a large series of 65 compounds that incorporated several aspects of structural variation has been synthesised through an efficient, divergent method that allowed for a number of analogues to be generated from common precursors. Subsequently, all analogues were assessed for their antibacterial activity against both Gram-positive ( Staphylococcus aureus ) and Gram-negative ( Escherichia coli ) bacteria. Overall, several compounds exhibited comparable or better activity than that of pseudoceratidine 1 , and it was found that this class of compounds is generally more effective against Gram-positive than Gram-negative bacteria. Furthermore, altering several structural features allowed for the establishment of a comprehensive structure activity relationship (SAR), where it was concluded that several structural features are critical for potent anti-bacterial activity, including di-halogenation (preferable bromine, but chlorine is also effective) on the pyrrole ring, two pyrrolic units in the structure and with one or more secondary amines in the chain adjoining these units, with longer chains giving rise to better activities.

Background: Dimethyl sulfide (DMS) is a small sulfur-containing impact odorant, imparting distinctive positive and / or negative characters to food and beverages. In white wine, the presence of DMS at perception threshold is considered to be a fault, contributing strong odors reminiscent of asparagus, cooked cabbage, and creamed corn. The source of DMS in wine has long been associated with S-methyl-l-methionine (SMM), a derivative of the amino acid methionine, which is thought to break down into DMS through chemical degradation, particularly during wine ageing., Results: We developed and validated a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with a stable isotope dilution assay (SIDA) to measure SMM in grape juice and wine. The application of this new method for quantitating SMM, followed by the quantitation of DMS using headspace-solid phase micro-extraction coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS), confirmed that DMS can be produced in wine via the chemical breakdown of SMM to DMS, with greater degradation observed at 28 °C than at 14 °C. Further investigation into the role of grape juice and yeast strain on DMS formation revealed that the DMS produced from three different Sauvignon blanc grape juices, either from the SMM naturally present or SMM spiked at 50 mmol L -1 , was modulated depending on each of the four strains of Saccharomyces cerevisiae wine yeast used for fermentation., Conclusion: This study confirms the existence of a chemical pathway to the formation of DMS and reveals a yeast-mediated mechanism towards the formation of DMS from SMM during alcoholic fermentation. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

New evidence on the role of H2S as a gasotransmitter suggests that the true signalling effectors are polysulfides. Both oxidized polysulfides and hydropolysulfides were synthesized and their presence in S. cerevisiae was observed for the first time. A single gene-deletant approach allowed observation of the modulation of polysulfide species and levels.

3-(methylthio)-1-propanol (methionol), produced by yeast as an end-product of L-methionine (L-Met) catabolism, imparts off-odours reminiscent of cauliflower and potato to wine. Saccharomyces cerevisiae ARO genes, including transaminases Aro8p and Aro9p, and decarboxylase Aro10p, catalyse two key steps forming methionol via the Ehrlich pathway. We compared methionol concentrations in wines fermented by single Δaro8, Δaro9 and Δaro10 deletants in lab strain BY4743 versus wine strain Zymaflore F15, and F15 double- and triple-aro deletants versus single-aro deletants, using headspace-solid phase microextraction coupled with gas chromatography-mass spectrometry.Deletion of two or more aro genes increased growth lag phase, with the greatest delay exhibited by F15 Δaro8 Δaro9. The single Δaro8 deletion decreased methionol by 44% in BY4743 and 92% in F15, while the Δaro9 deletion increased methionol by 46% in F15 but not BY4743. Single deletion of Δaro10 had no effect on methionol.Unexpectedly, F15 Δaro8 Δaro9 and F15 Δaro8 Δaro9 Δaro10 produced more methionol than F15 Δaro8. In the absence of Aro8p and Aro9p, other transaminases may compensate or an alternative pathway may convert methanethiol to methionol. Our results confirm that Ehrlich pathway genes differ greatly between lab and wine yeast strains, impacting downstream products such as methionol.

The aldehyde 3-S-glutathionylhexanal is an intermediate which is produced during the formation of the wine aroma precursor 3-S-glutathionylhexanol, after the reaction of glutathione with trans-2-hexenal. This study was conducted to assess whether the chemical, as opposed to the enzymatic, production of 3-S-glutathionylhexanal could occur at a significant rate in grape juice. LC-MS/MS was used in low- and high-resolution modes, in combination with functional group derivatization, to identify and quantitate products. In comparison to cysteine, glutathione was found to induce less cyclized products on reaction with trans-2-alkanals and the glutathione-derived products were more reactive to hydrogen sulfite. The zero-order rates for 3-S-glutathionylhexanal formation in model grape juice were 1.08 ± 0.08 and 0.45 ± 0.05 mg/(L·day) glutathione equivalents at 25 and 13 °C, respectively, and the reaction rate increased 3-fold by increasing the pH from 3.2 to 3.8. 3-S-Glutathionylhexanal was detected in all five white grape juices examined. The concentration of the aldehyde could be increased by up to 10-fold after being released from hydrogen sulfite, demonstrating a potentially novel source for the production of varietal thiol aroma compounds in wine.

Spores are the most resistant form of microbial cells, thus difficult to inactivate. The pathogenic or food spoilage effects of certain spore-forming microorganisms have been the primary basis of sterilization and pasteurization processes. Thermal sterilization is the most common method to inactivate spores present on medical equipment and foods. High pressure processing (HPP) is an emerging and commercial non-thermal food pasteurization technique. Although previous studies demonstrated the effectiveness of thermal and non-thermal spore inactivation, the in-depth mechanisms of spore inactivation are as yet unclear. Live and dead forms of two food spoilage bacteria, a mould and a yeast were examined using scanning electron microscopy before and after the inactivation treatment. Alicyclobacillus acidoterrestris and Geobacillus stearothermophilus bacteria are indicators of acidic foods pasteurization and sterilization processes, respectively. Neosartorya fischeri is a phyto-pathogenic mould attacking fruits. Saccharomyces cerevisiae is a yeast with various applications for winemaking, brewing, baking and the production of biofuel from crops (e.g. sugar cane). Spores of the four microbial species were thermally inactivated. Spores of S. cerevisiae were observed in the ascus and free form after thermal and HPP treatments. Different forms of damage and cell destruction were observed for each microbial spore. Thermal treatment inactivated bacterial spores of A. acidoterrestris and G. stearothermophilus by attacking the inner core of the spore. The heat first altered the membrane permeability allowing the release of intracellular components. Subsequently, hydration of spores, physicochemical modifications of proteins, flattening and formation of indentations occurred, with subsequent spore death. Regarding N. fischeri, thermal inactivation caused cell destruction and leakage of intracellular components. Both thermal and HPP treatments of S. cerevisiae free spores attacked the inner membrane, altering its permeability, and allowing in final stages the transfer of intracellular components to the outside. The spore destruction caused by thermal treatment was more severe than HPP, as HPP had less effect on the spore core. All injured spores have undergone irreversible volume and shape changes. While some of the leakage of spore contents is visible around the deformed but fully shaped spore, other spores exhibited large indentations and were completely deformed, apparently without any contents inside. This current study contributed to the understanding of spore inactivation by thermal and non-thermal processes., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Sauvignon blanc wine, balanced by herbaceous and tropical aromas, is fermented at low temperatures (10-15 °C). Anecdotal accounts from winemakers suggest that cold fermentations produce and retain more "fruity" aroma compounds; nonetheless, studies have not confirmed why low temperatures are optimal for Sauvignon blanc. Thirty-two aroma compounds were quantitated from two Marlborough Sauvignon blanc juices fermented at 12.5 and 25 °C, using Saccharomyces cerevisiae strains EC1118, L-1528, M2, and X5. Fourteen compounds were responsible for driving differences in aroma chemistry. The 12.5 °C-fermented wines had lower 3-mercaptohexan-1-ol (3MH) and higher alcohols but increased fruity acetate esters. However, a sensory panel did not find a significant difference between fruitiness in 75% of wine pairs based on fermentation temperature, in spite of chemical differences. For wine pairs with significant differences (25%), the 25 °C-fermented wines were fruitier than the 12.5 °C-fermented wines, with high fruitiness associated with 3MH. We propose that the benefits of low fermentation temperatures are not derived from increased fruitiness but a better balance between fruitiness and greenness. Even so, since 75% of wines showed no significant difference, higher fermentation temperatures could be utilized without detriment, lowering costs for the wine industry.

Low fermentation temperatures are of importance to food and beverage industries working with Saccharomyces cerevisiae Therefore, the identification of genes demonstrating a positive impact on fermentation kinetics is of significant interest. A set of 121 mapped F 1 progeny, derived from a cross between haploid strains BY4716 (a derivative of the laboratory yeast S288C) and wine yeast RM11-1a, were fermented in New Zealand Sauvignon Blanc grape juice at 12.5°. Analyses of five key fermentation kinetic parameters among the F 1 progeny identified a quantitative trait locus (QTL) on chromosome I with a significant degree of linkage to maximal fermentation rate ( V max ) at low temperature. Independent deletions of two candidate genes within the region, FLO1 and SWH1 , were constructed in the parental strains (with S288C representing BY4716). Fermentation of wild-type and deletion strains at 12.5 and 25° confirmed that the genetic linkage to V max corresponds to the S288C version of the FLO1 allele, as the absence of this allele reduced V max by ∼50% at 12.5°, but not at 25°. Reciprocal hemizygosity analysis (RHA) between S288C and RM11-1a FLO1 alleles did not confirm the prediction that the S288C version of FLO1 was promoting more rapid fermentation in the opposing strain background, suggesting that the positive effect on V max derived from S288C FLO1 may only provide an advantage in haploids, or is dependent on strain-specific cis or trans effects. This research adds to the growing body of evidence demonstrating the role of FLO1 in providing stress tolerance to S. cerevisiae during fermentation., (Copyright © 2017 Deed et al.)

Although the yeast response to low temperature has industrial significance for baking, lager brewing and white wine fermentation, the molecular response of yeast cells to low temperature remains poorly characterised. Transcriptional changes were quantified in a commercial wine yeast, Enoferm M2, fermented at optimal (25 °C) and low temperature (12.5 °C), at two time points during fermentation of Sauvignon blanc grape juice. The transition from early to mid-late fermentation was notably less severe in the cold than at 25 °C, and the Rim15p-Gis1p pathway was involved in effecting this transition. Genes for three key nutrients were strongly influenced by low temperature fermentation: nitrogen, sulfur and iron/copper, along with changes in the cell wall and stress response. Transcriptional analyses during wine fermentation at 12.5 °C in four F1 hybrids of M2 also highlighted the importance of genes involved in nutrient utilisation and the stress response. We identified transcription factors that may be important for these differences between genetic backgrounds. Since low fermentation temperatures cause fundamental changes in membrane kinetics and cellular metabolism, an understanding of the physiological and genetic limitations on cellular performance will assist breeding of improved industrial strains.