The influence of Torulaspora delbrueckii on beer fermentation.

In this study, the effect of five different Torulaspora delbrueckii strains in combination with an ale type Saccharomyces cerevisiae on physical, chemical, microbiological, aroma composition, and sensory profiles of beer were examined. The ethyl alcohol content of produced beers ranged from 5.46% (v/v) to 5.93% (v/v), while the highest alcohol amount was obtained using a pure culture of S. cerevisiae. The major volatiles among beer aroma compounds was acetaldehyde, n-propanol, 3-methyl-butanol, 2-methyl-butanol, ethyl acetate, isoamyl acetate, 2,3-butanedione, and 2,3-pentanedione. It was ascertained that the total amount of higher alcohols was higher in the S. cerevisiae control beer compared to all mixed fermentations. Total ester levels were higher in all the mixed culture beers than the control beer. Sensory evaluation showed that all the mixed cultures of S. cerevisiae and T. delbrueckii positively influenced the sensory profile of the beers. Strain Y1031 was the most preferred and was characterized as rich in hop aroma and full bodied. It is therefore a suitable strategy to use T. delbrueckii in mixed fermentations with S. cerevisiae to produce beer with a distinctive flavor. The results demonstrate that, T. delbrueckii strains isolated or commercialized for winemaking can be equally employed as well in brewing.

Antifungal activity of non-conventional yeasts against Botrytis cinerea and non-Botrytis grape bunch rot fungi.

Grapes harbour a plethora of non-conventional yeast species. Over the past two decades, several of the species have been extensively characterised and their contribution to wine quality is better understood. Beyond fermentation, some of the species have been investigated for their potential as alternative biological tools to reduce grape and wine spoilage. However, such studies remain limited to a few genera. This work aimed to evaluate the antagonistic activity of grape must-derived non-conventional yeasts against Botrytis cinerea and non-Botrytis bunch-rotting moulds and to further elucidate mechanisms conferring antifungal activity. A total of 31 yeast strains representing 21 species were screened on different agar media using a dual culture technique and liquid mixed cultures, respectively. Pichia kudriavzevii was the most potent with a minimum inhibitory concentration of 102 cells/mL against B. cinerea but it had a narrow activity spectrum. Twelve of the yeast strains displayed broad antagonistic activity, inhibiting three strains of B. cinerea (B05. 10, IWBT FF1 and IWBT FF2), a strain of Aspergillus niger and Alternaria alternata. Production of chitinases and glucanases in the presence of B. cinerea was a common feature in most of the antagonists. Volatile and non-volatile compounds produced by antagonistic yeast strains in the presence of B. cinerea were analysed and identified using gas and liquid chromatography mass spectrometry, respectively. The volatile compounds identified belonged mainly to higher alcohols, esters, organosulfur compounds and monoterpenes while the non-volatile compounds were cyclic peptides and diketopiperazine. To our knowledge, this is the first report to demonstrate inhibitory effect of the non-volatile compounds produced by various yeast species.

Non-Saccharomyces yeast for lowering wine alcohol levels: partial aeration versus standard conditions.

Non-Saccharomyces yeasts have been suggested for use in wine production for lowering alcohol content. In this study, 23 non-Saccharomyces yeasts were investigated in laboratory-scale trials using previously frozen grape must. Both aerated and standard fermentation conditions were investigated and the fermentations were co-inoculated with a commercial Saccharomyces cerevisiae reference yeast strain. Sugar consumed for percentage alcohol formed was calculated from sugar and alcohol measurements. The non-Saccharomyces yeasts showed greater variability in sugar consumption compared with the S. cerevisiae reference yeast. Two of the yeast strains (Starmerella bacillaris and Wickerhamomyces anomalus) consumed more sugar than the S. cerevisiae reference yeast under the same conditions. These two strains were subsequently used in a small-scale wine production trial following a similar aeration and standard fermentation strategy. The wine production trials using aeration compared with the standard strategy showed shorter fermentation times, increased biomass formation and more sugar utilized for alcohol produced, but reduced wine quality. The same yeasts under standard fermentation conditions also showed increased use of sugar, but neutral or positive effects on wine quality. The S. bacillaris strain showed the most potential for use in wine production for lowering alcohol content.

Effect of alcoholic and acetous fermentations on the phenolic acids of Kei-apple (Dovyalis caffra L.) fruit.

BACKGROUND: The Kei apple is a tree found on the African continent. Limited information exists on the effect of alcoholic and acetous fermentation on the phytochemicals of Kei apple. The fruit has increased concentrations of l-malic, ascorbic, and phenolic acids among other compounds. Juice was co-inoculated with Schizosaccharomyces pombe (Sp) and Saccharomyces cerevisiae (Sc) to induce alcoholic fermentation (AF). Acetous fermentation followed AF, using an acetic acid bacteria (AAB) consortium. RESULTS: Saccharomyces cerevisiae + Sp wines and vinegars had the highest pH. Total acidity, soluble solids and l-malic acid decreased during AF and acetous fermentation, and was highest in Sc wines and vinegars. Volatile acidity (VA) concentration was highest in Sp vinegars but was not significantly different from Sc and Sc + Sp vinegars. Gallic acid was highest in Sp wines and vinegars, whereas syringic acid was highest in Sc wines and vinegars. The Sc + Sp wines were highest in caffeic, p-coumaric, and protocatechuic acids. Schizosaccharomyces pombe vinegars were highest in caffeic and p-coumaric acids. Highest concentrations of ferulic and sinapic acids were found in Sp and Sc wines, respectively. Chlorogenic acid was most abundant phenolic acid in both wines and vinegars. CONCLUSION: Saccharomyces cerevisiae + Sp and Sc fermentation had a positive effect on most phenolic acids; Sc + AAB had an increased effect on syringic and chlorogenic acids, whereas Sp + AAB resulted in an increase in gallic, caffeic, and p-coumaric acids. The AAB selected had minimal performance with respect to VA production in comparison to commercial vinegars. Acetic acid bacteria selection for acetous fermentation should therefore be reconsidered and the decrease of certain phenolic acids during acetous fermentation needs to be investigated. © 2021 Society of Chemical Industry.

The Use of Candida pyralidae and Pichia kluyveri to Control Spoilage Microorganisms of Raw Fruits Used for Beverage Production.

Undesired fermentation of fruit-derived beverages by fungal, yeast and bacterial spoilage organisms are among the major contributors of product losses in the food industry. As an alternative to chemical preservatives, the use of Candida pyralidae and Pichia kluyveri was assessed for antimicrobial activity against several yeasts (Dekkera bruxellensis, Dekkera anomala, Zygosaccharomyces bailii) and fungi (Botrytis cinerea, Colletotrichum acutatum and Rhizopus stolonifer) associated with spoilage of fruit and fruit-derived beverages. The antagonistic properties of C. pyralidae and P. kluyveri were evaluated on cheap solidified medium (grape pomace extract) as well as on fruits (grapes and apples). Volatile organic compounds (VOCs) from C. pyralidae and P. kluyveri deemed to have antimicrobial activity were identified by gas chromatography-mass spectrometry (GC-MS). A cell suspension of C. pyralidae and P. kluyveri showed growth inhibition activity against all spoilage microorganisms studied. Direct contact and extracellular VOCs were two of the mechanisms of inhibition. Twenty-five VOCs belonging to the categories of alcohols, organic acids and esters were identified as potential sources for the biocontrol activity observed in this study. This study reports, for the first time, the ability of C. pyralidae to inhibit fungal growth and also for P. kluyveri to show growth inhibition activity against spoilage organisms (n = 6) in a single study.

Aeration, Agitation and Cell Immobilization on Corncobs and Oak Wood Chips Effects on Balsamic-Styled Vinegar Production.

Optimum fermentor conditions are essential for desired microbial growth and activity in fermentations. In balsamic vinegar fermentation systems, the microorganisms used must endure several stressful conditions including high sugar concentration, low water activity, high osmotic pressure and high acetic acid concentration. Consequently, the present study was aimed at improving the performance of a microbial consortium of non-Saccharomyces yeast and acetic acid bacteria during balsamic-styled vinegar fermentation. Cell immobilization via adsorption on corncobs and oak wood chips in combination with aeration and agitation effects, have never been tested during balsamic-styled vinegar fermentation. Therefore, fermentations were initially conducted under static conditions without aeration with successive fermentations also being subjected to low (0.15 vvm min-1) and high (0.3 vvm min-1) aeration. The results showed improved acetification rates when cells were immobilized on corncobs under static conditions. Low aeration showed better acetification rates (1.45-1.56 g·L·day-1), while only free-floating cells were able to complete fermentations (1.2 g·L·day-1) under high aeration conditions. Overall, cells immobilized on corncobs showed higher acetification rates of 1.56 and 2.7 g·L·day-1 under low aeration and static fermentations, respectively. Oak wood chips were determined to be less efficient adsorbents due to their relatively smooth surface, while the rough surface and porosity of corncobs led to improved adsorption and, therefore, enhanced acetification rates.

Characterisation of hybrid yeasts for the production of varietal Sauvignon blanc wine - A review.

The wine yeast Saccharomyces cerevisiae forms an integral part of wine production by converting relatively 'neutral' flavoured grape must into varietal aromatic wines. Additionally, non-Saccharomyces strains can be used with S. cerevisiae for the production of wines with more complexity. Yeast strains, to varying extents, produce and/or mediate the release of a whole range of key metabolites, which in turn contribute to enhanced aroma and flavour of the final wine, especially Sauvignon blanc. These metabolites viz. thiols are dependent on yeast-expressed enzymes during fermentation. Inoculation with an appropriate yeast will, therefore, lead to more commercial wine sales due to resultant wines with sought-after aroma and flavour. Likewise, inoculation with the incorrect yeast will have a negative effect on sales. It is also important to have quality control measures in place to ensure that the inoculated yeast strain quickly dominate, and is present throughout the fermentation process. Traditionally, the laborious contour clamped homogeneous electric field (CHEF) DNA karyotyping technique was shown to be reliable in this regard, however cutting-edge matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF MS) biotyping is proving to be a faster alternative. As both methods have advantages and disadvantages, they should be used in complementary as opposed to competitively. Standard chemical and descriptive sensory analyses of wine also serve as evaluation and/or characterisation tools of yeast starter cultures. Additionally, metabolomic and proteomic profiling using gas chromatography (GC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) of final wines was shown to be instrumental yeast evaluation tools. Therefore, this review highlights the importance and practicality of a more inclusive approach to evaluate and characterise novel yeast strains used for winemaking by deploying traditional and modern chemical and organoleptic evaluation techniques of wines in conjunction with cutting-edge omics approaches towards enhancing white wine varietal aroma.

Grape Pomace Extracts as Fermentation Medium for the Production of Potential Biopreservation Compounds.

Microbial spoilage causes food losses in the food industry and as such, the use of synthetic chemical preservatives is still required. The current study proposes the use of agro-waste, i.e., grape pomace extracts (GPE), as production medium for biopreservation compounds. Production kinetics, subsequent to optimization using response surface methodology (RSM) for biopreservation compounds production was studied for three yeasts using GPE broth as a fermentation medium. The results showed that the highest volumetric zone of inhibition (VZI) was 1.24 L contaminated solidified media (CSM) per mL biopreservation compounds used (BCU) when Candida pyralidae Y1117 was inoculated in a pH 3-diluted GPE broth (150 g L-1) incubated at 25 °C for 24 h. Similar conditions were applied for Pichia kluyveri Y1125 and P. kluyveri Y1164, albeit under slightly elongated fermentation periods (up to 28 h), prior to the attainment of a maximum VZI of only 0.72 and 0.76 L CSM mL-1 ACU, respectively. The potential biopreservation compounds produced were identified to be isoamyl acetate, isoamyl alcohol, 2-phenyl ethylacetate and 2-phenyl ethanol.

Anthocyanins and polyphenols in Cabernet Franc wines produced with Saccharomyces cerevisiae and Torulaspora delbrueckii yeast strains: Spectrophotometric analysis and effect on selected sensory attributes.

Grapes and wine contain phenolics divided into non-flavonoid and flavonoid classes. Yeast modulates the phenolics of wine by adsorption onto yeast cell walls. This may be advantageous for colour and quality. The effect of Torulaspora delbrueckii (654&M2/1) and Saccharomyces cerevisiae (VIN13) on phenolics and sensory attributes of Cabernet Franc wines (2012/2013) were evaluated. Spectrophotometric analysis was used to determine the phenolic content. Sensory analysis was conducted by expert tasters to evaluate the wines. ANOVA showed that polyphenols and anthocyanins were higher in M2/1 + VIN13 wines (2012/2013) than 654 + VIN13 wines. Colour, fruitiness, mouthfeel, sweetness, astringency and quality were different between treatments. 654 + VIN13 wines (2012) were higher in polyphenols, acidity, astringency and mouthfeel than M2/1 + VIN13 wines. M2/1 + VIN13 (2012/2013) had increased colour and quality than 654 + VIN13 wines. Two treatments were identified for Cabernet Franc wines; M2/1 + VIN13 and 654 + VIN13, which resulted in wines with increased colour and wines with increased mouthfeel and astringency, respectively.

Mating of natural Saccharomyces cerevisiae strains for improved glucose fermentation and lignocellulosic inhibitor tolerance.

Natural Saccharomyces cerevisiae isolates from vineyards in the Western Cape, South Africa were evaluated for ethanol production in industrial conditions associated with the production of second-generation biofuels. The strains displayed high phenotypic diversity including the ability to grow at 45 °C and in the presence of 20% (v/v) ethanol, strain YI13. Strains HR4 and YI30 were inhibitor-tolerant under aerobic and oxygen-limited conditions, respectively. Spore-to-spore hybridization generated progeny that displayed heterosis, including increased ethanol productivity and improved growth in the presence of a synthetic inhibitor cocktail. Hybrid strains HR4/YI30#6 and V3/YI30#6 were able to grow at a high salt concentration (2 mol/L NaCl) with V3/YI30#6 also able to grow at a high temperature (45 °C). Strains HR4/YI30#1 and #3 were inhibitor-tolerant, with strain HR4/YI30#3 having similar productivity (0.36 ± 0.0036 g/L per h) as the superior parental strain, YI30 (0.35 ± 0.0058 g/L per h). This study indicates that natural S. cerevisiae strains display phenotypic variation and heterosis can be achieved through spore-to-spore hybridization. Several of the phenotypes (temperature-, osmo-, and inhibitor tolerance) displayed by both the natural strains and the generated progeny were at the maximum conditions reported for S. cerevisiae strains.

Influence of Lachancea thermotolerans on cv. Emir wine fermentation.

The present paper describes the behaviour of Lachancea thermotolerans and Saccharomyces cerevisiae in pure, co-cultured and sequential fermentations in cv. Emir grape must. Faster fermentation rates were observed in wine made with a pure culture of S. cerevisiae and wine produced with simultaneously inoculated cultures of L. thermotolerans and S. cerevisiae. Both L. thermotolerans and S. cerevisiae gave high population numbers. The use of L. thermotolerans in mixed and sequential cultures led to an increase in final total acidity content in the wines, varying in the range 5.40-6.28 g/l (as tartaric acid), compared to pure culture S. cerevisiae, which gave the lowest level of total acidity (5 g/l). The increase was in the order of 1.18-2.06 g/l total acidity. Increase in final acidity by the use of L. thermotolerans might be useful to improve wines with low acidity due to global climate change. Volatile acidity levels (as acetic acid) were in the range 0.53-0.73 g/l, while the concentration of ethyl alcohol varied in the range 10.76-11.62% v/v. Sequential fermentations of wines and pure culture fermentation of L. thermotolerans resulted in reduction in the concentrations of acetaldehyde and higher alcohols, with exception of N-propanol and esters. According to the sensory analysis, wine obtained with sequential inoculation of L. thermotolerans followed by inoculation of S. cerevisiae after 24 h, and simultaneous inoculation of these yeasts, was the most preferred. Copyright © 2016 John Wiley & Sons, Ltd.

Not your ordinary yeast: non-Saccharomyces yeasts in wine production uncovered.

Saccharomyces cerevisiae and grape juice are 'natural companions' and make a happy wine marriage. However, this relationship can be enriched by allowing 'wild' non-Saccharomyces yeast to participate in a sequential manner in the early phases of grape must fermentation. However, such a triangular relationship is complex and can only be taken to 'the next level' if there are no spoilage yeast present and if the 'wine yeast' - S. cerevisiae - is able to exert its dominance in time to successfully complete the alcoholic fermentation. Winemakers apply various 'matchmaking' strategies (e.g. cellar hygiene, pH, SO2 , temperature and nutrient management) to keep 'spoilers' (e.g. Dekkera bruxellensis) at bay, and allow 'compatible' wild yeast (e.g. Torulaspora delbrueckii, Pichia kluyveri, Lachancea thermotolerans and Candida/Metschnikowia pulcherrima) to harmonize with potent S. cerevisiae wine yeast and bring the best out in wine. Mismatching can lead to a 'two is company, three is a crowd' scenario. More than 40 of the 1500 known yeast species have been isolated from grape must. In this article, we review the specific flavour-active characteristics of those non-Saccharomyces species that might play a positive role in both spontaneous and inoculated wine ferments. We seek to present 'single-species' and 'multi-species' ferments in a new light and a new context, and we raise important questions about the direction of mixed-fermentation research to address market trends regarding so-called 'natural' wines. This review also highlights that, despite the fact that most frontier research and technological developments are often focussed primarily on S. cerevisiae, non-Saccharomyces research can benefit from the techniques and knowledge developed by research on the former.

Characterisation of commercial and natural Torulaspora delbrueckii wine yeast strains.

Forty-three South African Torulaspora delbrueckii yeast isolates from the ARC Infruitec-Nietvoorbij yeast culture collection, the T. delbrueckii type strain (CBS 1146), one reference T. delbrueckii strain (CBS 4663), two T. delbrueckii strains isolated from commercial yeast blends (Viniflora® Harmony.nsac and Viniflora® Melody.nsac), and a commercial Saccharomyces cerevisiae yeast (VIN 13) had their identities confirmed and were characterised using conventional and molecular microbiological techniques. These included a selection of growth media as well as CHEF electrophoretic karyotyping and PCR-RFLP analyses. Based on the biochemical and physiological results the strains were divided into 13 groups. The performances of the yeasts were also monitored by means of laboratory-scale fermentations in grape must at 15 °C and 22 °C. The fermentation kinetic data showed that at 22 °C, the yeasts were divided into two distinct groups, a faster and a slower fermenting group. The fermentation curves of the laboratory-scale study at 15 °C showed that, at this lower temperature, the yeasts also fermented at different speeds, but the fermentation curves showed greater separation. The biochemical and physiological grouping did not coincide with the fermentation abilities and good fermenters could be found in more than one group. Chemical analyses of the resultant wines (alcohol, volatile acidity, glycerol, total SO2, residual sugar) were used in Principle Component Analyses. The yeasts that grouped close to the S. cerevisiae reference strain (VIN 13) showed more acceptable wine chemical profiles, while those further away displayed less acceptable profiles. Three locally isolated strains and one commercial T. delbrueckii yeast strain, Viniflora® Harmony.nsac. produced wines with acceptable chemical profiles at both temperatures. These strains also had comparable fermentation kinetics to the S. cerevisiae reference. Therefore, depending on the fermentation temperature, different T. delbrueckii strains will be suitable for specific wine styles and some may even be considered for single inoculations without S. cerevisiae in industrial fermentations.

The effect of scale on gene expression: commercial versus laboratory wine fermentations.

Molecular and cellular processes that are responsible for industrially relevant phenotypes of fermenting microorganisms are a central focus of biotechnological research. Such research intends to generate insights and solutions for fermentation-based industries with regards to issues such as improving product yield or the quality of the final fermentation product. For logistical reasons, and to ensure data reproducibility, such research is mostly carried out in defined or synthetic media and in small-scale fermentation vessels. Two questions are frequently raised regarding the applicability of this approach to solve problems experienced in industrial fermentations: (1) Is synthetic medium a sufficiently accurate approximation of the generally more complex natural (and frequently highly variable) substrates that are employed in most fermentation-based industries, and (2) can results obtained in small-scale laboratory fermentations be extrapolated to large-scale industrial environments? Here, we address the second question through a comparative transcriptomic approach by assessing the response of an industrial wine yeast strain fermenting a natural grape juice in small-scale laboratory and large-scale industrial conditions. In yeast, transcriptome analysis is arguably the best available tool to holistically assess the physiological state of a population and its response to changing environmental conditions. The data suggest that scale does indeed impact on some environmental parameters such as oxygen availability. However, the data show that small-scale fermentations nevertheless accurately reflect general molecular processes and adaptations during large-scale fermentation and that extrapolation of laboratory datasets to real industrial processes can be justified.

A Geographically Diverse Collection of Schizosaccharomyces pombe Isolates Shows Limited Phenotypic Variation but Extensive Karyotypic Diversity.

The fission yeast Schizosaccharomyces pombe has been widely used to study eukaryotic cell biology, but almost all of this work has used derivatives of a single strain. We have studied 81 independent natural isolates and 3 designated laboratory strains of Schizosaccharomyces pombe. Schizosaccharomyces pombe varies significantly in size but shows only limited variation in proliferation in different environments compared with Saccharomyces cerevisiae. Nucleotide diversity, π, at a near neutral site, the central core of the centromere of chromosome II is approximately 0.7%. Approximately 20% of the isolates showed karyotypic rearrangements as detected by pulsed field gel electrophoresis and filter hybridization analysis. One translocation, found in 6 different isolates, including the type strain, has a geographically widespread distribution and a unique haplotype and may be a marker of an incipient speciation event. All of the other translocations are unique. Exploitation of this karyotypic diversity may cast new light on both the biology of telomeres and centromeres and on isolating mechanisms in single-celled eukaryotes.