Alcoholic Fermentation Research Articles

The Effect of the Addition of baker’s Yeast and Tape Yeast with the Length of Fermentation on the Quality of Carica vinegar

Abstract Vinegar is a condiment produced by alcoholic and acetic acid fermentation of various sugary or starchy ingredients. Vinegar can be produced from a substrate containing a certain amount of ethanol which comes from various sources such as coconut water, pineapple peel, fruit and coffee grounds. Acetic acid can be produced by two methods, namely synthetic/chemical and microbiological or fermentation. Vinegar is widely available in the market, is commonly consumed and contains chemicals that are harmful to the body. Vinegar is produced through an industrial process with a pure chemical synthesis process. In this study, the basic ingredients for the process of making acetic acid came from carica fruit juice. The stages of forming acetic acid include the addition of sugar, the first fermentation and the second fermentation. In the first fermentation process, the yeast used is baker’s yeast, while the second fermentation uses tape yeast. Determination of variables in this study using a factorial design. The results of the study obtained the optimum effect of fermentation time, yeast mass and volume of alcohol for the manufacture of carica fruit vinegar. The higher the concentration of the tape yeast used in the processing of carica vinegar, the higher the alcohol content and total acid produced, and the lower the pH and sugar content. Of all the treatments the best results were produced from an experiment with a total acid of 5.2. The experimental variable was a yeast mass of 4 g, a fermentation time of 11 days and a volume of alcohol used was 30 ml.

Yeast as a cell factory for fermentative production of ethanol from xylose

BackgroundXylose is the most plentiful pentose sugar found in the lignocellulosic biomass in hemicellulose fractions. This pentose sugar has high potential to use as raw material for bioethanol production, as hemicellulose fractions account for 20–30 % in the lignocellulose biomass. However, bioconversion of xylose into ethanol is challenging for industrial prospective a sonly few microbes encompass the pentose fermentation pathway. MethodsBioconversion of xylose into bioethanol using yeasts is preferred since these organisms are more resistant and have higher ethanol tolerance limit compared to bacteria. During fermentation process xylose is not readily consumed by the microbes like glucose. Metabolic engineering has so far been unsuccessful to furnish adequate outcomes; and one of the reasons of this is the lack of consideration of a range of environmental factors which play potential role fermentative ethanol production form xylose. Significant FindingsThis review focuses on the bioethanol production from xylose highlighting the metabolic engineering segment, when microbes enter into fermenting phase. On this basis, the best possible technology can be evolved for subsequent commercialization of the product.

Gaining insights into the responses of individual yeast cells to ethanol fermentation using Raman tweezers and chemometrics

Saccharomyces cerevisiae is the most common microbe used for the industrial production of bioethanol, and it encounters various stresses that inhibit cell growth and metabolism during fermentation. However, little is currently known about the physiological changes that occur in individual yeast cells during ethanol fermentation. Therefore, in this work, Raman spectroscopy and chemometric techniques were employed to monitor the metabolic changes of individual yeast cells at distinct stages during high gravity ethanol fermentation. Raman tweezers was used to acquire the Raman spectra of individual yeast cells. Multivariate curve resolution-alternating least squares (MCR-ALS) and principal component analysis were employed to analyze the Raman spectra dataset. MCR-ALS extracted the spectra of proteins, phospholipids, and triacylglycerols and their relative contents in individual cells. Changes in intracellular biomolecules showed that yeast cells undergo three distinct physiological stages during fermentation. In addition, heterogeneity among yeast cells significantly increased in the late fermentation period, and different yeast cells may respond to ethanol stress via different mechanisms. Our findings suggest that the combination of Raman tweezers and chemometrics approaches allows for characterizing the dynamics of molecular components within individual cells. This approach can serve as a valuable tool in investigating the resistance mechanism and metabolic heterogeneity of yeast cells during ethanol fermentation.

Extensive remodeling of sugar metabolism through gene loss and horizontal gene transfer in a eukaryotic lineage

BackgroundIn yeasts belonging to the subphylum Saccharomycotina, genes encoding components of the main metabolic pathways, like alcoholic fermentation, are usually conserved. However, in fructophilic species belonging to the floral Wickerhamiella and Starmerella genera (W/S clade), alcoholic fermentation was uniquely shaped by events of gene loss and horizontal gene transfer (HGT).ResultsBecause HGT and gene losses were first identified when only eight W/S-clade genomes were available, we collected publicly available genome data and sequenced the genomes of 36 additional species. A total of 63 genomes, representing most of the species described in the clade, were included in the analyses. Firstly, we inferred the phylogenomic tree of the clade and inspected the genomes for the presence of HGT-derived genes involved in fructophily and alcoholic fermentation. We predicted nine independent HGT events and several instances of secondary loss pertaining to both pathways. To investigate the possible links between gene loss and acquisition events and evolution of sugar metabolism, we conducted phenotypic characterization of 42 W/S-clade species including estimates of sugar consumption rates and fermentation byproduct formation. In some instances, the reconciliation of genotypes and phenotypes yielded unexpected results, such as the discovery of fructophily in the absence of the cornerstone gene (FFZ1) and robust alcoholic fermentation in the absence of the respective canonical pathway.ConclusionsThese observations suggest that reinstatement of alcoholic fermentation in the W/S clade triggered a surge of innovation that goes beyond the utilization of xenologous enzymes, with fructose metabolism playing a key role.

Influence of different stress factors during the elaboration of grape must's pieddecuve on the dynamics of yeast populations during alcoholic fermentation

The pieddecuve (PdC) technique involves using a portion of grape must to undergo spontaneous fermentation, which is then used to inoculate a larger volume of must. This allows for promoting autochthonous yeasts present in the must, which can respect the typicality of the resulting wine. However, the real impact of this practice on the yeast population has not been properly evaluated. In this study, we examined the effects of sulphur dioxide (SO2), temperature, ethanol supplementation, and time on the dynamics and selection of yeasts during spontaneous fermentation to be used as PdC. The experimentation was conducted in a synthetic medium and sterile must using a multi-species yeast consortium and in un-inoculated natural grape must. Saccharomyces cerevisiae dominated both the PdC and fermentations inoculated with commercial wine yeast, displaying similar population growth regardless of the tested conditions. However, using 40 mg/L of SO2 and 1% (v/v) ethanol during spontaneous fermentation of Muscat of Alexandria must allowed the non-Saccharomyces to be dominant during the first stages, regardless of the temperature tested. These findings suggest that it is possible to apply the studied parameters to modulate the yeast population during spontaneous fermentation while confirming the effectiveness of the PdC methodology in controlling alcoholic fermentation.

Effect of Cap Management Frequency on the Phenolic, Chromatic, and Sensory Composition of Cabernet Sauvignon Wines from the Central Coast of California over Two Vintages.

Cabernet Sauvignon from the California Paso Robles AVA was processed with a contrasting array of cap management frequencies, consisting of punch-down (PD) frequencies (0, 1, 2, and 3 PD/day) over two vintages, one of which the fruit was harvested at two contrasting maturity levels. Wines followed with up to 3 years of bottle aging for basic and phenolic chemistry, and the wines of the second harvest of 2020 were submitted to sensory analysis. There were almost non-existent effects due to the frequency of punch downs on parameters such as ethanol, pH, titratable acidity, lactic acid, and glucose + fructose. In 2019, the chromatic differences between different PD regimes were subtle, and minor effects of the punch-down frequency were observed for tannins and total phenolics. During the early stages of alcoholic fermentation, higher levels of all anthocyanin classes were observed in 1 PD wines and the lowest levels in 0 PD wines. The anthocyanin content of the wines of the first harvest (unripe) was 27% higher than that of the wines of the second harvest (ripe), but these differences disappeared after 3 years of bottle aging irrespective of the vintage and harvest date. Acylated anthocyanins were preferentially lost during aging, especially in 2019 wines and, to a lesser extent, in 2020 wines. In 2020, the polymeric pigment content of the wines of the second harvest was higher than in the wines of the first harvest, with 3 PD wines showing higher polymeric pigments and yellow hues than 0 and 2 PD wines after 3 years of bottle aging. Sensory analysis of the second harvest of the 2020 wines showed that the wines of all four PD regimes were perceived as drying, signifying they were perceived as equally astringent, which is consistent with comparable tannin levels on said wines. The perception of bitterness increased with the frequency of punch downs; thus, 3 PD wines showed the highest bitterness perception. It was concluded that in sufficiently warm fermentations and small volumes, phenolic extraction occurs regardless of fruit maturity and under conditions of minimum mixing.

Comparison of Different Spectral Ranges to Monitor Alcoholic and Acetic Fermentation of Red Grape Must Using FT-NIR Spectroscopy and PLS Regression

AbstractWine vinegar is produced through a two-phase fermentation of grape must: initially, yeast converts grape sugars into ethanol, and subsequently, acetobacteria oxidize ethanol into acetic acid. This process, spanning weeks when conducted by surface fermentation, requires constant monitoring of ethanol and total acidity levels. To enhance the quality and efficiency of process monitoring, vinegar production is shifting to faster, environmentally sustainable methods. Near-infrared (NIR) spectroscopy, recognized for its non-invasiveness and speed, is ideal for online implementation in process control. This study tracked dual fermentation in red grape must over an extended period, monitoring two different batches simultaneously to assess fermentation kinetics and reproducibility. Ethanol content and total acidity were analyzed in fermenting musts throughout the whole fermentation process using both classical laboratory analyses and FT-NIR spectroscopy. Principal Component Analysis (PCA) was used to explore the spectral dataset, then Partial Least Squares (PLS) was used to develop calibration models for predicting ethanol and acidity. The models calculated considering the entire spectral range were compared with those obtained for two narrower zones, where more cost-effective and easily miniaturizable sensors are available on the market. FT-NIR allowed to effectively determine ethanol content and acidity (R2Pred > 0.98), both over the entire range (12,500–4000 cm−1, corresponding to 800–2500 nm) and in the 10,526–6060 cm−1 (950–1650 nm) region. Although less satisfactory, still acceptable results were obtained in the 12,500–9346 cm−1 (800–1070 nm) region (R2Pred > 0.81), confirming the potential for cost-effective devices in real-time fermentation monitoring.

Fermentation Analytical Technology (FAT): Monitoring industrial E. coli fermentations using absolute quantitative 1H NMR spectroscopy

BackgroundTo perform fast, reproducible, and absolute quantitative measurements in an automated manner has become of paramount importance when monitoring industrial processes, including fermentations. Due to its numerous advantages – including its inherent quantitative nature – Proton Nuclear Magnetic Resonance (1H NMR) spectroscopy provides an ideal tool for the time-resolved monitoring of fermentations. However, analytical conditions, including non-automated sample preparation and long relaxation times (T1) of some metabolites, can significantly lengthen the experimental time and make implementation in an industrial set up unfeasible. ResultsWe present a high throughput method based on Standard Operating Procedures (SOPs) and 1H NMR, which lays the foundation for what we call Fermentation Analytical Technology (FAT). Our method was developed for the accurate absolute quantification of metabolites produced during Escherichia coli industrial fermentations. The method includes: (1) a stopped flow system for non-invasive sample collection followed by sample quenching, (2) automatic robot-assisted sample preparation, (3) fast 1H NMR measurements, (4) metabolites quantification using multivariate curve resolution (MCR), and (5) metabolites absolute quantitation using a novel correction factor (k) to compensate for the short recycle delay (D1) employed in the 1H NMR measurements. The quantification performance was tested using two sample types: buffer solutions of chemical standards and real fermentation samples. Five metabolites – glucose, acetate, alanine, phenylalanine and betaine – were quantified. Absolute quantitation ranged between 0.64 and 3.40 mM in pure buffer, and 0.71–7.76 mM in real samples. SignificanceThe proposed method is generic and can be straight forward implemented to other types of fermentations, such as lactic acid, ethanol and acetic acid fermentations. It provides a high throughput automated solution for monitoring fermentation processes and for quality control through absolute quantification of key metabolites in fermentation broth. It can be easily implemented in an at-line industrial setting, facilitating the optimization of the manufacturing process towards higher yields and more efficient and sustainable use of resources.

Bioinspired Colorimetric Double Inverse Opal Photonic Crystal Indicators for Ethanol Concentration Sensing in Fermentation Engineering.

Photonic crystal-based ethanol concentration indicators with rapid response and brilliant structural color output definitely take a place in colorimetric sensors. Here, based on the H-bond-regulated swelling of acrylate shape memory polymers (SMPs) and the solvent-induced structural color change of the double inverse opal photonic crystals (DIOPCs), new-type photonic crystals (PCs) colorimetric indicators were constructed, exhibiting a span of maximum reflection wavelength (λmax) up to ∼166 nm in response to alcohols with concentrations from 0 to 100 vol %. DIOPC indicators (DIOPCIs) show a rapid response to alcohols (<1.5 s) and output different structural colors (covering from blue to red). The colorimetric sensing mechanism includes the solvent-triggered recovery of the inverse opal skeleton, the cosolvency effect and H-bonds induced swelling/shrinkage of the polymer, the phase separation between polystyrene (PS) microsphere and polymer skeleton, and the light diffraction of DIOPCs. While ensuring a larger λmax span by regulating the H-bond interactions in polymer chains through acrylamide (AAm), AAm-modified DIOPCIs are sensitive to some specific ethanol concentrations. The real-time sensing of ethanol concentration during fermentation verified the practicability of DIOPCIs, thus establishing a visual model between structural color and corresponding fermentation kinetics. We envisage that the DIOPCIs will contribute to the intelligentization of the alcoholic fermentation and distillation industry.

Transcriptomic and metabolomic correlation analysis: effect of initial SO2 addition on higher alcohol synthesis in Saccharomyces cerevisiae and identification of key regulatory genes.

Higher alcohols are volatile compounds produced during alcoholic fermentation that affect the quality and safety of the final product. This study used a correlation analysis of transcriptomics and metabolomics to study the impact of the initial addition of SO2 (30, 60, and 90 mg/L) on the synthesis of higher alcohols in Saccharomyces cerevisiae EC1118a and to identify key genes and metabolic pathways involved in their metabolism. Transcriptomics and metabolomics correlation analyses were performed and differentially expressed genes (DEGs) and differential metabolites were identified. Single-gene knockouts for targeting genes of important pathways were generated to study the roles of key genes involved in the regulation of higher alcohol production. We found that, as the SO2 concentration increased, the production of total higher alcohols showed an overall trend of first increasing and then decreasing. Multi-omics correlation analysis revealed that the addition of SO2 affected carbon metabolism (ko01200), pyruvate metabolism (ko00620), glycolysis/gluconeogenesis (ko00010), the pentose phosphate pathway (ko00030), and other metabolic pathways, thereby changing the precursor substances. The availability of SO2 indirectly affects the formation of higher alcohols. In addition, excessive SO2 affected the growth of the strain, leading to the emergence of a lag phase. We screened the ten most likely genes and constructed recombinant strains to evaluate the impact of each gene on the formation of higher alcohols. The results showed that ADH4, SER33, and GDH2 are important genes of alcohol metabolism in S. cerevisiae. The isoamyl alcohol content of the EC1118a-ADH4 strain decreased by 21.003%; The isobutanol content of the EC1118a-SER33 strain was reduced by 71.346%; and the 2-phenylethanol content of EC1118a-GDH2 strain was reduced by 25.198%. This study lays a theoretical foundation for investigating the mechanism of initial addition of SO2 in the synthesis of higher alcohols in S. cerevisiae, uncovering DEGs and key metabolic pathways related to the synthesis of higher alcohols, and provides guidance for regulating these mechanisms.

Evaluation of the effect of Torulaspora delbrueckii on important volatile compounds in navel orange original brandy using E–nose combined with HS–SPME–GC–MS

Simultaneous inoculation of non-Saccharomyces cerevisiae during the alcoholic fermentation process has been found to be an effective strategy for enhancing wine flavor. This study aimed to investigate the effect of Torulaspora delbrueckii NCUF305.2 on the flavor of navel orange original brandy (NOOB) using E–nose combined with HS–SPME–GC–MS. The results showed a significant increase (p < 0.05) in the sensitivity of NOOB to W5C, W3C, W1S, and W3S sensors by mixed fermentation (MF). Esters in NOOB increased by 4.13%, while higher alcohols increased by 21.93% (p < 0.001), terpenes and others increased by 52.07% and 40.99% (p < 0.01), respectively. Notably, several important volatile compounds with relative odor activity values above 10 showed an increase. Sensory analysis revealed that a more pronounced citrus-like flavor and higher overall appearance scores were found in MF than in pure fermentation (PF). These findings offer valuable theoretical guidance for enhancing the quality of fruit brandies.

Application of essential oils as natural antimicrobials in lactic acid bacteria contaminating fermentation for the production of organic cachaça

Sugarcane-based fermentation is an essential process for different sectors of economic importance, such as the food industry with fermented and distilled beverages. However, this process can suffer from high contamination by wild yeasts and bacteria, especially lactic acid bacteria (LAB). This makes it necessary to use decontamination strategies and search for new methods that have a low environmental impact and contribute to the production of organic products. Among the options, oregano and thyme essential oils stand out for their antibacterial compounds. The aim of this study was to use oregano and thyme essential oils as natural antimicrobials in the alcoholic fermentation of sugar cane juice. Initially, the minimum inhibitory concentration of the essential oils in the fermentation was assessed through turbidity in the sensitivity test, which allowed us to determine which concentrations of essential oils would inhibit the contaminants, 3 morphologically selected LAB strains, as well as assessing the viability of CA-11. For LAB, 3 concentrations of each essential oil were tested, ranging from 0.1 to 0.4 μl/mL, while for CA-11 it was 0.06, to 0.1 μl/mL. The results indicated a maximum total value of essential oils per ml of 0.06 μl. Based on this result, a 24-1 fractional factorial was established, with 8 conditions +3 central points, with 4 variables, oregano essential oil (0, 0.03 and 0.06 μl/ml), thyme (0, 0.03 and 0.06 μl/ml), initial soluble solids (14, 16 and 18°Brix) and initial yeast concentration (2.5, 3 and 3.5 g/l), with the temperature set at 32 °C for a period of 12 h. The results showed that the center point condition with 0.03 μl/ml of oregano EO, 0.03 μl/ml of thyme controlled the proliferation of contaminating bacteria compared to the control condition. In the experimental validation, the treatment with essential oils had a lower final population of LAB (5.95 log) than the final population of the control treatment (6.53 log), and it was also observed that the treatment with EOs had an alcohol production around 3 % higher than the treatment without antimicrobials. The experimental validation phase confirmed the synergistic action of oregano and thyme essential oils in controlling the proliferation of contaminating bacteria. In conclusion, it was possible to determine the synergistic antimicrobial action of essential oils against LAB during alcoholic fermentation based on organic sugar cane.

Effect of co-inoculation of Saccharomyces and non-Saccharomyces yeasts and nutrients addition during malolactic fermentation on apple cider composition

Apple cider production is a complex fermentation process that involves the conversion of apple juice into cider through the actions of yeast and malolactic bacteria. This study aimed to investigate the effects of the co-inoculation of Saccharomyces and non-Saccharomyces yeasts, as well as nutrient supplementation in the presence of malolactic bacteria Oenoccocus oeni on the composition of apple cider. The amino acid profile of the samples was not significantly influenced by the addition of lactic bacteria or nutrients in cider processing. The highest ester content was recorded in the single yeast strain fermented ciders (55.32%–88.13%). This increase was visible also for the malolactic ciders, independent from nutrient supplementation. Co-inoculated Saccharomyces and Torulaspora delbrueckii ciders showed increased higher alcohols contents, both after the alcoholic (42.66%–53.51%) and malolactic fermentations (42.81%–65.94%). Following the hydroxycynnamic acids class (141.47–327.44 mg/L), flavonoids (114.81–253.44 mg/L) were the second most abundant class of phenolic compounds in ciders, which generally registered a considerable increase following malolactic fermentation. The sour/acidic taste was the most prevalent sensory attribute of the cider. Lower values were recorded for sweet, bitter, and astringent. The strongest apple aroma was perceived in the co-inoculated Saccharomyces and T. delbrueckii yeast strains. The co-fermentation of Saccharomyces and Torulaspora delbrueckii yeasts led to increased levels of desirable aroma compounds, contributing to a more complex and appealing sensory profile, while the nutrient supplementation provided an improved malolactic fermentation efficiency and overall cider quality.

Transcriptome and metabolome analyses reveal molecular insights into waterlogging tolerance in Barley

Waterlogging stress is one of the major abiotic stresses affecting the productivity and quality of many crops worldwide. However, the mechanisms of waterlogging tolerance are still elusive in barley. In this study, we identify key differentially expressed genes (DEGs) and differential metabolites (DM) that mediate distinct waterlogging tolerance strategies in leaf and root of two barley varieties with contrasting waterlogging tolerance under different waterlogging treatments. Transcriptome profiling revealed that the response of roots was more distinct than that of leaves in both varieties, in which the number of downregulated genes in roots was 7.41-fold higher than that in leaves of waterlogging sensitive variety after 72 h of waterlogging stress. We also found the number of waterlogging stress-induced upregulated DEGs in the waterlogging tolerant variety was higher than that of the waterlogging sensitive variety in both leaves and roots in 1 h and 72 h treatment. This suggested the waterlogging tolerant variety may respond more quickly to waterlogging stress. Meanwhile, phenylpropanoid biosynthesis pathway was identified to play critical roles in waterlogging tolerant variety by improving cell wall biogenesis and peroxidase activity through DEGs such as Peroxidase (PERs) and Cinnamoyl-CoA reductases (CCRs) to improve resistance to waterlogging. Based on metabolomic and transcriptomic analysis, we found the waterlogging tolerant variety can better alleviate the energy deficiency via higher sugar content, reduced lactate accumulation, and improved ethanol fermentation activity compared to the waterlogging sensitive variety. In summary, our results provide waterlogging tolerance strategies in barley to guide the development of elite genetic resources towards waterlogging-tolerant crop varieties.

Key role of K+ and Ca2+ in high-yield ethanol production by S. Cerevisiae from concentrated sugarcane molasses

BackgroundSaccharomyces cerevisiae is an important microorganism in ethanol synthesis, and with sugarcane molasses as the feedstock, ethanol is being synthesized sustainably to meet growing demands. However, high-concentration ethanol fermentation based on high-concentration sugarcane molasses—which is needed for reduced energy consumption of ethanol distillation at industrial scale—is yet to be achieved.ResultsIn the present study, to identify the main limiting factors of this process, adaptive laboratory evolution and high-throughput screening (Py-Fe3+) based on ARTP (atmospheric and room-temperature plasma) mutagenesis were applied. We identified high osmotic pressure, high temperature, high alcohol levels, and high concentrations of K+, Ca2+, K+ and Ca2+ (K+&Ca2+), and sugarcane molasses as the main limiting factors. The robust S. cerevisiae strains of NGT-F1, NGW-F1, NGC-F1, NGK+, NGCa2+ NGK+&Ca2+-F1, and NGTM-F1 exhibited high tolerance to the respective limiting factor and exhibited increased yield. Subsequently, ethanol synthesis, cell morphology, comparative genomics, and gene ontology (GO) enrichment analysis were performed in a molasses broth containing 250 g/L total fermentable sugars (TFS). Additionally, S. cerevisiae NGTM-F1 was used with 250 g/L (TFS) sugarcane molasses to synthesize ethanol in a 5-L fermenter, giving a yield of 111.65 g/L, the conversion of sugar to alcohol reached 95.53%. It is the highest level of physical mutagenesis yield at present.ConclusionOur results showed that K+ and Ca2+ ions primarily limited the efficient production of ethanol. Then, subsequent comparative transcriptomic GO and pathway analyses showed that the co-presence of K+ and Ca2+ exerted the most prominent limitation on efficient ethanol production. The results of this study might prove useful by promoting the development and utilization of green fuel bio-manufactured from molasses.

Copper-based grape pest management has impacted wine aroma

Despite the high energetic cost of the reduction of sulfate to H2S, required for the synthesis of sulfur-containing amino acids, some wine Saccharomyces cerevisiae strains have been reported to produce excessive amounts of H2S during alcoholic fermentation, which is detrimental to wine quality. Surprisingly, in the presence of sulfite, used as a preservative, wine strains produce more H2S than wild (oak) or wine velum (flor) isolates during fermentation. Since copper resistance caused by the amplification of the sulfur rich protein Cup1p is a specific adaptation trait of wine strains, we analyzed the link between copper resistance mechanism, sulfur metabolism and H2S production. We show that a higher content of copper in the must increases the production of H2S, and that SO2 increases the resistance to copper. Using a set of 51 strains we observed a positive and then negative relation between the number of copies of CUP1 and H2S production during fermentation. This complex pattern could be mimicked using a multicopy plasmid carrying CUP1, confirming the relation between copper resistance and H2S production. The massive use of copper for vine sanitary management has led to the selection of resistant strains at the cost of a metabolic tradeoff: the overproduction of H2S, resulting in a decrease in wine quality.

Periodic on-off operations of the self-cycling ethanol fermentation process: Stability analysis and optimal operation strategies

Fuel ethanol is highlighted for being renewable and carbon neutral, but a multi-scale challenge exists that hampers its role as a future energy substitute, and with the breakthrough of cellulose pretreatment and enzyme hydrolysis technology, fermentation itself has become one of the main limiting factors for process enhancement. To tackle the problem, this paper studies periodic operation strategies to improve process performance and avoid possible instabilities. Firstly, superiority by periodic operation over the corresponding steady-state operation is verified. Then, to settle the problem of possible instabilities under impulsive control, stability of state-impulsive control is conducted and for the studied case, any self-cycling fermentation starts from batch mode with 50 % discharge-and-refill portion exhibits superior repeatability property, which allows continuous fermentation to operate under very slow overall dilution rate to cope with the extreme slow fermentation rate. Finally, on-off operation problem is formulated that is governed by a multidimensional nonlinear system, affine to the manipulated variable that is under the isoperimetric constraint; with the use of Pontryagin maximum principle, the optimal synthesis of the optimal strategies under integral constraint is computed out, and a practically plausible on-off operation strategy for ethanol fermentation is formulated with guaranteed periodic stability and optimality.

Overview of the distinctive characteristics of strawberry, raspberry, and blueberry in berries, berry wines, and berry spirits.

Red berries have gained popularity as functional and nutritious food due to their health benefits, leading to increased consumer demand and higher production, totaling over 11,000ktons for strawberries, raspberries, and blueberries combined in 2021. Nutritionally, strawberries, raspberries, and blueberries present high levels of vitamin C (9.7-58.8mg/100g dry weight [dw]), folates (6-24µg/100g dw), and minerals (96-228mg/100g dw). Due to their perishable nature, producers have utilized alcoholic fermentation to extend their shelf life, not only increasing the lifespan of red berries but also attracting consumers through the production of novel beverages. Strawberry, blueberry, and raspberry wines possess low alcohol (5.5-11.1% v/v), high acidity (3.2-17.6g/L), and interesting bioactive molecules such as phenolic compounds, carotenoids, polysaccharides, and melatonin. Distillation holds tremendous potential for reducing food waste by creating red berry spirits of exceptional quality. Although research on red berry spirits is still in the early stages, future studies should focus on their production and characterization. By incorporating these factors, the production chain would become more sustainable, profitable, and efficient by reducing food waste, capitalizing on consumer acceptance, and leveraging the natural health-promoting characteristics of these products. Therefore, this review aims to provide a comprehensive overview of the characteristics of strawberry, blueberry, and red raspberry in berries, wines, and spirits, with a focus on their chemical composition and production methods.

The Value of Indigenous &lt;i&gt;Saccharomyces cerevisiae&lt;/i&gt; Strains as Useful Enzymes Producers: A Systematic Review

This systematic review has focused on the selection and characterization of indigenous Saccharomyces cerevisiae strains for enzymes' production obtained from the fermentation process. The primary studies assessed have demonstrated that indigenous S. cerevisiae strains produce a wide range of enzymes during fermentation. These strains have shown different characteristics and were isolated from various locations, including those belonging to the main components of the wine production chain. The research methodology applied to find primary studies about the optimal conditions for enzyme production was effective and identified a wide range of enzyme classes. Cultivation media amended with natural or synthetic inducer substrates were found to be suitable carbon and/or nitrogen sources for producing enzymes of interest in plate-based screening. In submerged fermentation studies, the use of agro-industrial residues has been identified as a relevant carbon source for microbe development and metabolism. For the indigenous S. cerevisiae strains, the majority of the enzymes found belong to the glucosidases and hydrolases classes. Due to the activity of these enzymes, yeasts have been traditionally used in classic biotechnological applications, particularly in the process of alcoholic fermentations. Those enzymes are responsible for producing metabolites (such as aromatic compounds) that contribute to the desirable aroma and/or taste during the fermentation of wine, as well as other fermented or distilled beverages. The methods used for enzyme screening were of quantitative and/or qualitative nature and were carried out under acidic pH conditions (3.5-6.5) and at temperatures of 28-35 ℃. The results of this systematic review have shown that the enzymes from indigenous S. cerevisiae are highly relevant in various biotechnological applications, particularly in the food industry. Conclusions and future perspectives of our findings were discussed in terms of potential developments of novel application methods of the indigenous yeasts and their enzymes in agriculture and industry.

Tannin-Tolerant Saccharomyces cerevisiae Isolated from Traditional Fermented Tea Leaf (Miang) and Application in Fruit Wine Fermentation Using Longan Juice Mixed with Seed Extract as Substrate.

This study focused on isolating tannin-tolerant yeasts from Miang, a fermented tea leaf product collected from northern Laos PDR, and investigating related food applications. From 43 Miang samples, six yeast isolates capable of ethanol production were obtained, with five isolates showing growth on YPD agar containing 4% (w/v) tannic acid. Molecular identification revealed three isolates as Saccharomyces cerevisiae (B5-1, B5-2, and C6-3), along with Candida tropicalis and Kazachstania humilis. Due to safety considerations, only Saccharomyces spp. were selected for further tannic acid tolerance study to advance food applications. Tannic acid at 1% (w/v) significantly influenced ethanol fermentation in all S. cerevisiae isolates. Notably, B5-2 and C6-3 showed high ethanol fermentation efficiency (2.5% w/v), while others were strongly inhibited. The application of tannin-tolerant yeasts in longan fruit wine (LFW) fermentation with longan seed extract (LSE) supplementation as a source of tannin revealed that C6-3 had the best efficacy for LFW fermentation. C6-3 showed promising efficacy, particularly with LSE supplementation, enhancing phenolic compounds, antioxidant activity, and inhibiting α-glucosidase activity, indicating potential antidiabetic properties. These findings underscore the potential of tannin-tolerant S. cerevisiae C6-3 for fermenting beverages from tannin-rich substrates like LSE, with implications for functional foods and nutraceuticals promoting health benefits.