Saccharomyces Cerevisiae Strains Research Articles

Isolation, identification, and tolerance analysis of yeast during the natural fermentation process of Sidamo coffee beans.

Yeast, which plays a pivotal role in the brewing, food, and medical industries, exhibits a close relationship with human beings. In this study, we isolated and purified 60 yeast strains from the natural fermentation broth of Sidamo coffee beans to screen for indigenous beneficial yeasts. Among them, 25 strains were obtained through morphological characterization on nutritional agar medium from Wallerstein Laboratory (WL), with molecular biology identifying Saccharomyces cerevisiae strain YBB-47 and the remaining 24 yeast strains identified as Pichia kudriavzevii. We investigated the fermentation performance, alcohol tolerance, SO2 tolerance, pH tolerance, sugar tolerance, temperature tolerance, ester production capacity, ethanol production capacity, H2S production capacity, and other brewing characteristics of YBB-33 and YBB-47. The results demonstrated that both strains could tolerate up to 3% alcohol by volume at a high sucrose mass concentration (400g/L) under elevated temperature conditions (40 ℃), while also exhibiting a remarkable ability to withstand an SO2 mass concentration of 300g/L at pH 3.2. Moreover, S. cerevisiae YBB-47 displayed a rapid gas production rate and strong ethanol productivity. whereas P. kudriavzevii YBB-33 exhibited excellent alcohol tolerance. Furthermore, this systematic classification and characterization of coffee bean yeast strains from the Sidamo region can potentially uncover additional yeasts that offer high-quality resources for industrial-scale coffee bean production.

Effects of ultrasound and microwave treatments on oenological yeast microbiota: comparison with a flash pasteurisation

SummaryThe control of yeast biota in wine could be a challenge, as different genera could coexist with different effects on wine quality; generally, SO2 plays an important role in this context, while other approaches have been proposed. In this article, two Microwave (MW) and two Ultrasound treatments (US) were tested and compared to a flash pasteurisation on seven yeast strains (Saccharomyces cerevisiae, Saccharomyces bayanus, Brettanomyces spp., Schizosaccharomyces pombe, Hanseniaspora osmophila, Zygosaccharomyces bailii, Pichia membranifaciens); viable count (viability) and yeast growth kinetic (vitality) were assessed. The treatments reduced viable count by 27%, with a higher biocidal effect of US, depending on yeast and treatment conditions; they also impaired yeast growth causing a delay, which is a significant increase of the lag phase, while the effect on the growth rate or the maximum OD attained in the steady state was less significant. In conclusion, the US generally exerted a stronger effect than MW and flash pasteurisation and Schizo. pombe and Z. bailii were the less resistant yeasts. A partial uncoupling was found between vitality and viability for Z. bailii, while in Schizo. pombe the delay in the growth was generally related to the decrease in the cell count immediately after the treatment.

Coupled engineering strategy of CYB2 deletion and ACS1 overexpression improves cellulosic lactic acid production by Saccharomyces cerevisiae

Lactic acid plays a significant role in the food industry as a natural preservative and serves as a platform chemical for the production of biodegradable poly-lactic acid. While its sustainable production through fermentation using cellulosic biomass instead of traditional sugar and starch raw materials shows promise, there remains limited information available on this method. In this study, a xylose-fermenting and lactic acid-tolerant Saccharomyces cerevisiae strain (BK01) was engineered for improved cellulosic lactic acid production using the promising energy crop, kenaf. Specifically, CYB2 was deleted to inhibit lactic acid consumption (BK02), which was further engineered by ACS1 overexpression to remove the cytotoxic acetate derived from cellulosic biomass (BK03). The BK03 produced 3.25-fold more lactic acid from kenaf pulp hydrolysate than the parental strain (BK01). During pH-controlled fermentation, lactic acid production by BK03 reached a titer of 31 g/L and a productivity of 1.30 g/L-h, the highest value reported so far. This is one of the few studies on yeast-based lactic acid production from lignocellulose.

Optimal trade-off between boosted tolerance and growth fitness during adaptive evolution of yeast to ethanol shocks

BackgroundThe selection of Saccharomyces cerevisiae strains with higher alcohol tolerance can potentially increase the industrial production of ethanol fuel. However, the design of selection protocols to obtain bioethanol yeasts with higher alcohol tolerance poses the challenge of improving industrial strains that are already robust to high ethanol levels. Furthermore, yeasts subjected to mutagenesis and selection, or laboratory evolution, often present adaptation trade-offs wherein higher stress tolerance is attained at the expense of growth and fermentation performance. Although these undesirable side effects are often associated with acute selection regimes, the utility of using harsh ethanol treatments to obtain robust ethanologenic yeasts still has not been fully investigated.ResultsWe conducted an adaptive laboratory evolution by challenging four populations (P1–P4) of the Brazilian bioethanol yeast, Saccharomyces cerevisiae PE-2_H4, through 68–82 cycles of 2-h ethanol shocks (19–30% v/v) and outgrowths. Colonies isolated from the final evolved populations (P1c–P4c) were subjected to whole-genome sequencing, revealing mutations in genes enriched for the cAMP/PKA and trehalose degradation pathways. Fitness analyses of the isolated clones P1c–P3c and reverse-engineered strains demonstrated that mutations were primarily selected for cell viability under ethanol stress, at the cost of decreased growth rates in cultures with or without ethanol. Under this selection regime for stress survival, the population P4 evolved a protective snowflake phenotype resulting from BUD3 disruption. Despite marked adaptation trade-offs, the combination of reverse-engineered mutations cyr1A1474T/usv1Δ conferred 5.46% higher fitness than the parental PE-2_H4 for propagation in 8% (v/v) ethanol, with only a 1.07% fitness cost in a culture medium without alcohol. The cyr1A1474T/usv1Δ strain and evolved P1c displayed robust fermentations of sugarcane molasses using cell recycling and sulfuric acid treatments, mimicking Brazilian bioethanol production.ConclusionsOur study combined genomic, mutational, and fitness analyses to understand the genetic underpinnings of yeast evolution to ethanol shocks. Although fitness analyses revealed that most evolved mutations impose a cost for cell propagation, combination of key mutations cyr1A1474T/usv1Δ endowed yeasts with higher tolerance for growth in the presence of ethanol. Moreover, alleles selected for acute stress survival comprising the P1c genotype conferred stress tolerance and optimal performance under conditions simulating the Brazilian industrial ethanol production.

Response mechanisms of different Saccharomyces cerevisiae strains to succinic acid

BackgroundThe production of succinic acid (SA) from biomass has attracted worldwide interest. Saccharomyces cerevisiae is preferred for SA production due to its strong tolerance to low pH conditions, ease of genetic manipulation, and extensive application in industrial processes. However, when compared with bacterial producers, the SA titers and productivities achieved by engineered S. cerevisiae strains were relatively low. To develop efficient SA-producing strains, it’s necessary to clearly understand how S. cerevisiae cells respond to SA.ResultsIn this study, we cultivated five S. cerevisiae strains with different genetic backgrounds under different concentrations of SA. Among them, KF7 and NBRC1958 demonstrated high tolerance to SA, whereas NBRC2018 displayed the least tolerance. Therefore, these three strains were chosen to study how S. cerevisiae responds to SA. Under a concentration of 20 g/L SA, only a few differentially expressed genes were observed in three strains. At the higher concentration of 60 g/L SA, the response mechanisms of the three strains diverged notably. For KF7, genes involved in the glyoxylate cycle were significantly downregulated, whereas genes involved in gluconeogenesis, the pentose phosphate pathway, protein folding, and meiosis were significantly upregulated. For NBRC1958, genes related to the biosynthesis of vitamin B6, thiamin, and purine were significantly downregulated, whereas genes related to protein folding, toxin efflux, and cell wall remodeling were significantly upregulated. For NBRC2018, there was a significant upregulation of genes connected to the pentose phosphate pathway, gluconeogenesis, fatty acid utilization, and protein folding, except for the small heat shock protein gene HSP26. Overexpression of HSP26 and HSP42 notably enhanced the cell growth of NBRC1958 both in the presence and absence of SA.ConclusionsThe inherent activities of small heat shock proteins, the levels of acetyl-CoA and the strains’ potential capacity to consume SA all seem to affect the responses and tolerances of S. cerevisiae strains to SA. These factors should be taken into consideration when choosing host strains for SA production. This study provides a theoretical basis and identifies potential host strains for the development of robust and efficient SA-producing strains.

193 The effects of mannan oligosaccharides on mortality and full value nursery swine

Abstract Mannan oligosaccharides [Actigen; ACT (Alltech, Inc., Nicholasville, KY)] are refined low molecular weight carbohydrates obtained from the cell wall of a specific strain of Saccharomyces cerevisiae. Peer-reviewed studies with nursery swine has suggested that ACT can positively impact intestinal morphology and permeability, inhibit gut colonization of enteric pathogens, and improve the pattern of immune responses with and without viral challenges. To validate these findings, three commercial scale experiments involving a total of 10,634 nursery pigs [PIC 337 × 1050, initially 6.4 kg of body weight (BW)] were conducted. Pens of 36 to 50 mixed sex pigs were randomly assigned to 1 of 2 dietary treatments supplemented with 0 or 800 ppm of ACT, with a total of 112 replications per treatment. In all trials, pigs were fed a nursery program comprised of two phases. Phase 1 from d 0 to 21 (experimental diets) and Phase 2 from d 22-37, 22-43, and 22-44, in Exp. 1, 2, and 3, respectively (common diets). Phase 1 diets were corn-soybean meal based and contained 9% whey permeate, 5% rolled oats, and were formulated to contain 1.38% SID Lysine. Phase 2 diets were corn-soybean meal based and formulated to contain 1.30% SID Lysine. In all experiments, data were analyzed using the GLIMMIX procedure of SAS (Cary, NC) in a randomized complete block design with pen serving as the experimental unit and initial BW serving as blocking factor. Percentage mortality and full value pig were analyzed using a binomial distribution. No significant ACT × experiment interactions (P > 0.10) were observed in mortality or full value pig (Table 1). From d 0 to 21, there was no evidence for differences in average daily gain (ADG), average daily feed intake (ADFI), or gain to feed ratio (G:F; P > 0.10) between dietary treatments (data not showed). From d 0 to 44, mortality was reduced (P < 0.0002), and full value pigs increased (P < 0.0004) in pigs fed diets with 800 ppm compared with pigs fed diets without ACT supplementation. In conclusion, whereas pigs fed diets supplemented with 800 ppm ACT did not show growth performance advantages, these experiments suggest livability and full value pig improvements.

Potential global cis and trans regulation of lncRNAs in Saccharomyces cerevisiae subjected to ethanol stress

Long noncoding RNAs (lncRNAs) are regulatory RNAs. Saccharomyces cerevisiae strains transcribe hundreds of lncRNAs. LncRNAs can regulate the expression of adjacent genes (cis-regulation) or distant genes from lncRNAs (trans-regulation). Here, we analyzed the potential global cis and trans-regulation of lncRNAs of yeast subjected to ethanol stress. For potential cis regulation, for BMA641-A and S288C strains, we observed that most lncRNA-neighbor gene pairs increased the expression at a certain point followed by a decrease, and vice versa. Based on the transcriptome profile and triple helix prediction between lncRNAs and promoters of coding genes, we observed nine different ways of potential trans regulation that work in a strain-specific manner. Our data provide an initial landscape of potential cis and trans regulation in yeast, which seems to be strain-specific.

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.

Utilizing deep ocean water in yeast fermentation for enhanced mineral-rich biomass production and fermentative regulation by proteomics modulation

Deep Ocean Water (DOW) is rich in minerals and serves as a natural source of nutrients. However, due to the inorganic nature of these minerals, cultivating yeast in DOW could aid in the fermentation process, and simultaneously, the yeast can assimilate the minerals from DOW, resulting in a mineral-enriched yeast biomass. Focusing on three DOW sources off the eastern coast of Taiwan (TT-1, HL-1, HL-2), we fermented various yeast strains of Saccharomyces cerevisiae. Therefore, this study investigates the effects of DOW on yeast growth, alcohol dehydrogenase activity, and the biological absorption of mineral ions by the yeast. Additionally, this research employs two-dimensional electrophoresis techniques to examine how the absorbed minerals influence the regulation of yeast proteins, thereby affecting biomass and metabolism. In the result, S. cerevisiae BCRC 21689 demonstrated a remarkable ability to bio-absorb minerals such as magnesium, calcium, potassium, and zinc from DOW, enhancing its growth and fermentation performance. Proteomic analysis revealed significant shifts in the expression of 21 proteins related to glycolytic and energy metabolism, alcohol metabolism, and growth regulation, all influenced by DOW's mineral-rich environment. This indicates that DOW's mineral content is a key factor in upregulating essential enzymes in glycolytic metabolism and alcohol dehydrogenase. An increase in proteins involved in synthesis and folding processes was also observed, leading to a substantial increase in yeast biomass. This study underscores the potential of DOW as a natural enhancer in yeast fermentation processes, enriching the yeast with diverse minerals and modulating proteomic expression to optimize yeast growth and fermentation.

Metabolomic analysis reveals the interactions between Chinese indigenous and commercial Saccharomyces cerevisiae strains during wine co-fermentations at low YAN concentration

The successful use of indigenous strains as starter cultures is dependent not only upon their capacity to complete a fermentation but also on their ability to dominate the fermentation. Chinese strain NX11424 was identified as an indigenous Chinese wine strain with commercial potential. Fermentations to assess the ability of NX11424 to compete with a commercial strain, Montrachet (UCDVEN 522), in pure and co-inoculations were performed with exo- and endo-metabolomic analyses. Metabolome analysis was run at three time points (days 2, 4 and 6). The metabolic profiles of all three treatment were similar at day 2. The endo-metabolome of the co-culture most closely resembles that of Montrachet strain at days 4 and 6, while exo-metabolomic data most closely resembles the NX11424 strain at day 6. The levels of key biomarkers among different fermentations varied at different time points, especially in amino acid metabolism; however glycerolipid, sucrose and starch metabolism were consistently most impactful in driving differences across treatment. Sulfur metabolism was also differential among treatments, with higher levels of associated metabolites in Montrachet strain. This study demonstrated the effects of pure and co-starter fermentations on cellular and wine metabolites, which can be helpful for control of wine fermentation and management of desirable wine flavor.

Saccharomyces cerevisiae oral immunization in mice using multi-antigen of the African swine fever virus elicits a robust immune response.

African swine fever virus (ASFV) is one of the most complex viruses. ASFV is a serious threat to the global swine industry because no commercial vaccines against this virus are currently available except in Vietnam. Moreover, ASFV is highly stable in the environment and can survive in water, feed, and aerosols for a long time. ASFV is transmitted through the digestive and respiratory tract. Mucosal immunity is the first line of defense against ASFV. Saccharomyces cerevisiae (SC), which has been certified by the U.S. Food and Drug Administration and has a generally recognized as safe status in the food industry, was used for oral immunization in this study. ASFV antigens were effectively expressed in recombinant SC strains with high DNA copy numbers and stable growth though surface display technology and chromosome engineering (δ-integration). The recombinant SC strains containing eight ASFV antigens-KP177R, E183L, E199L, CP204L, E248R, EP402R, B602L, and B646L- induced strong humoral and mucosal immune responses in mice. There was no antigenic competition, and these antigens induced Th1 and Th2 cellular immune responses. Therefore, the oral immunization strategy using recombinant SC strains containing multiple ASFV antigens demonstrate potential for future testing in swine, including challenge studies to evaluate its efficacy as a vaccine against ASFV.

The Value of Indigenous <i>Saccharomyces cerevisiae</i> 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.

Biosynthetic Pathways of Tryptophan Metabolites in Saccharomyces cerevisiae Strain: Insights and Implications.

Tryptophan metabolites, such as 5-hydroxytryptophan (5-HTP), serotonin, and melatonin, hold significant promise as supplements for managing various mood-related disorders, including depression and insomnia. However, their chemical production via chemical synthesis and phytochemical extraction presents drawbacks, such as the generation of toxic byproducts and low yields. In this study, we explore an alternative approach utilizing S. cerevisiae STG S101 for biosynthesis. Through a series of eleven experiments employing different combinations of tryptophan supplementation, Tween 20, and HEPES buffer, we investigated the production of these indolamines. The tryptophan metabolites were analyzed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Notably, setups replacing peptone in the YPD media with tryptophan (Run 3) and incorporating tryptophan along with 25 mM HEPES buffer (Run 4) demonstrated successful biosynthesis of 5-HTP and serotonin. The highest 5-HTP and serotonin concentrations were 58.9 ± 16.0 mg L-1 and 0.0650 ± 0.00211 mg L-1, respectively. Melatonin concentrations were undetected in all the setups. These findings underscore the potential of using probiotic yeast strains as a safer and conceivably more cost-effective alternative for indolamine synthesis. The utilization of probiotic strains presents a promising avenue, potentially offering scalability, sustainability, reduced environmental impact, and feasibility for large-scale production.

Autochthonous Ingredients for Craft Beer Production

Innovation in the beer market focuses on research into the different ingredients that make up this popular and sought-after beverage. Some of these innovations have focused on the use of autochthonous ingredients, which bring added value to products, reduce production costs and environmental impact, and provide beers with special organoleptic characteristics. This study focuses on the use of autochthonous ingredients from the Community of Madrid and the design of four recipes that are suited to these ingredients and consumer demand. In terms of the ingredients used, the micromalting of barley grown in the CM was carried out, while hops (Cascade variety for bittering hops and wild hops Rayo verde Tajuña and Torreta Tajuña for aroma) and yeast strains (Saccharomyces cerevisiae G 520 and G 354) were selected from previous studies, which were also sourced from the CM. For the barley malt, parameters such as protein content were determined, which was at the maximum limit but did not affect the final quality of the beers. The content of α- and β-acids in the hops was also determined, being higher in the Cascade variety. Different physicochemical analyses (GC, aromatic compounds, bitterness, colour, SO2, polyphenols, and antioxidant capacity) were carried out to determine the quality of the beers obtained from the four recipes designed, as well as tastings by a panel of experts and consumers. Finally, two recipes were rated the best by a panel of experts and consumers, where the cereal, hop, and fruit flavours stood out for their aromatic profile.

Screening of a Saccharomyces cerevisiae Strain with High 3-Methylthio-1-Propanol Yield and Optimization of Its Fermentation Conditions.

3-Methylthio-1-propanol (3-Met) is an important flavor compound in various alcoholic beverages such as Baijiu and Huangjiu. To maintain the content of 3-Met in these alcoholic beverages, it is necessary to screen a micro-organism with high yield of 3-Met from the brewing environment. In this study, the ability of yeast strains from the Baijiu brewing to produce 3-Met was analyzed, aiming to obtain yeast with high-yield 3-Met, and its fermentation conditions were optimized. Firstly, 39 yeast strains were screened using 3-Met conversion medium. The results showed that the majority of the strains from Baijiu brewing sources could produce 3-Met, and nearly half of the strains produced more than 0.5 g/L of 3-Met. Among these, yeast F10404, Y03401, and Y8#01, produced more than 1.0 g/L of 3-Met, with yeast Y03401 producing the highest amount at 1.30 g/L. Through morphological observation, physiological and biochemical analysis, and molecular biological identification, it was confirmed that yeast Y03401 was a Saccharomyces cerevisiae. Subsequently, the optimal fermentation conditions for 3-Met production by this yeast were obtained through single-factor designs, Plackett-Burman test, steepest ascent path design and response surface methodology. When the glucose concentration was 60 g/L, yeast extract concentration was 0.8 g/L, L-methionine concentration was 3.8 g/L, initial pH was 4, incubation time was 63 h, inoculum size was 1.6%, shaking speed was 150 rpm, loading volume was 50 mL/250 mL, and temperature was 26 °C, the content of 3-Met produced by S. cerevisiae Y03401 reached a high level of 3.66 g/L. It was also noteworthy that, in contrast to other study findings, this yeast was able to create substantial amounts of 3-Met even in the absence of L-methionine precursor. Based on the clear genome of S. cerevisiae and its characteristics in 3-Met production, S. cerevisiae Y03401 had broad prospects for application in alcoholic beverages such as Baijiu.

RT-qPCR based quantitative analysis of ARO and ADH genes in Saccharomyces cerevisiae and Metschnikowia pulcherrima strains growth white grape juice.

Yeast biosynthesizes fusel alcohols in fermentation through amino acid catabolism via the Ehrlich pathway. ARO8 and ARO9 genes are involved in the first step of the Ehrlich pathway, while ADH2 and ADH5 genes are involved in the last step. In this study, we describe RT-qPCR methods to determine the gene expression level of genes (ARO8, ARO9, ADH2, ADH5) found in Saccharomyces cerevisiae (Sc) and Metschnikowia pulcherrima (Mp) strains growth pasteurized white grape juice. We used RNA extraction and cDNA synthesis protocols. The RT-qPCR efficiency of primer pairs was evaluated by generating a standard curve through serial dilution of yeast-derived cDNA. Method performance criteria were determined for each RT-qPCR assay. Then, we evaluated the gene expression levels of the four genes in all samples. RNA extraction and cDNA synthesis from yeast samples demonstrated the method's capability to generate high-yield, high-purity nucleic acids, supporting further RT-qPCR analysis. The highest normalized gene expression levels of ARO8 and ARO9 were observed in SC1, SC4, and SC5 samples. No significant difference in ADH2 gene expression among Mp strains was observed during the examination of ADH2 and ADH5 genes (p < 0.05). We observed no expression of the ADH5 gene in Mp strains except MP6 strain. The expression of ADH2 and ADH5 genes was higher in Sc strains compared to Mp strains. The results suggest that the proposed RT-qPCR methods can measure gene expression of ARO8, ARO9, ADH2, and ADH5 in Sc and Mp strains growing in pasteurized white grape juice.

Development of freezing-resistant hybrid yeast from Saccharomyces cerevisiae for French bread dough

This study aimed to develop non-GMO freezing-resistant hybrid yeast for frozen French bread dough. The hybrids were generated by direct mating, using Saccharomyces cerevisiae strains from the ethanol industry (PE-2) and a baker's yeast (FLE). Four viable hybrids were generated and named F5P1, F5P33, F5P40, and F5P42. According to the 96-well microplate analyses, the growth curves of the hybrids after freezing and subsequent thawing presented similar behavior to those without freezing. In a performance test after the freezing step, F5P1 and F5P40 achieved a higher development index, with values of (147.1 ± 2.5) % and (102.1 ± 5.6) %, respectively. The peak reached by F5P1, after 50 min, was similar (p > 0.05) to that of FLE, which was at 110 min. The difference between the time required for F5P1 to achieve the growth peak with and without freezing was 10 min, whereas for FLE, this difference was three times higher, which highlights the improving freezing resistance of the hybrid. Therefore, the hybrid F5P1 showed a leavening capacity similar to FLE (p > 0.05) and improved freezing resistance, which indicates its potential for application to frozen French bread dough.

New Insights into the Production of Assyrtiko Wines from the Volcanic Terroir of Santorini Island Using Lachancea thermotolerans.

Assyrtiko is a rare ancient grape variety of Greece, which is known to produce Protected Designation of Origin (PDO) Santorini white wines. Besides the famous character of the volcanic terroir, Assyrtiko of Santorini is also marked by a low pH value and sharp acidity. The aim of the present study was to apply a new inoculation procedure that modulates the fermentation process by maintaining the unique sensorial characteristics of Assyrtiko wines based on acidity. For this purpose, the Lachancea thermotolerans species, known for the formation of lactic acid, was tested in sequential fermentation with three different Saccharomyces cerevisiae strains. At the end of the fermentation process, implantation control for S. cerevisiae strains (interdelta sequence profile analysis) was performed, oenological parameters were determined according to the OIV protocols, and the volatile compounds produced were measured by gas chromatography-mass spectrometry (GC/MS). Finally, all produced wines were evaluated by quantitative descriptive analysis by two groups of experts; the Greek team of oenologists from Santorini Island specialized in Assyrtiko wines, and the French team of oenologists specialized in wine from Bordeaux. As expected, the inoculated strain was the one that dominated the fermentation process, but nine S. cerevisiae indigenous strains were also identified in the produced wines. Lachancea thermotolerans produced 1 g/L of lactic and also modulated the volatile profile of the wines independently of the S. cerevisiae strain used. The origin of the panelists played an important role in bringing up sensorial traits, such as acidity. Our results led to a new interesting application of L. thermotolerans for white wine production adapted to climate change claims.

Enhancing xylose-fermentation capacity of engineered Saccharomyces cerevisiae by multistep evolutionary engineering in inhibitor-rich lignocellulose hydrolysate.

Major progress in developing Saccharomyces cerevisiae strains that utilize the pentose sugar xylose has been achieved. However, the high inhibitor content of lignocellulose hydrolysates still hinders efficient xylose fermentation, which remains a major obstacle for commercially viable second-generation bioethanol production. Further improvement of xylose utilization in inhibitor-rich lignocellulose hydrolysates remains highly challenging. In this work, we have developed a robust industrial S. cerevisiae strain able to efficiently ferment xylose in concentrated undetoxified lignocellulose hydrolysates. This was accomplished with novel multistep evolutionary engineering. First, a tetraploid strain was generated and evolved in xylose-enriched pretreated spruce biomass. The best evolved strain was sporulated to obtain a genetically diverse diploid population. The diploid strains were then screened in industrially relevant conditions. The best performing strain, MDS130, showed superior fermentation performance in three different lignocellulose hydrolysates. In concentrated corncob hydrolysate, with initial cell density of 1g DW/l, at 35°C, MDS130 completely coconsumed glucose and xylose, producing±7% v/v ethanol with a yield of 91% of the maximum theoretical value and an overall productivity of 1.22g/l/h. MDS130 has been developed from previous industrial yeast strains without applying external mutagenesis, minimizing the risk of negative side-effects on other commercially important properties and maximizing its potential for industrial application.

Valorisation of cotton post-industrial textile waste into lactic acid: chemo-mechanical pretreatment, separate hydrolysis and fermentation using engineered yeast

BackgroundThe textile industry has several negative impacts, mainly because it is based on a linear business model that depletes natural resources and produces excessive amounts of waste. Globally, about 75% of textile waste is disposed of in landfills and only 25% is reused or recycled, while less than 1% is recycled back into new garments. In this study, we explored the valorisation of cotton fabric waste from an apparel textile manufacturing company as valuable biomass to produce lactic acid, a versatile chemical building block.ResultsPost-industrial cotton patches were pre-treated with the aim of developing a methodology applicable to the industrial site involved. First, a mechanical shredding machine reduced the fabric into individual fibres of maximum 35 mm in length. Afterwards, an alkaline treatment was performed, using NaOH at different concentrations, including a 16% (w/v) NaOH enriched waste stream from the mercerisation of cotton fabrics. The combination of chemo-mechanical pre-treatment and enzymatic hydrolysis led to the maximum recovery yield of 90.46 ± 3.46%, corresponding to 74.96 ± 2.76 g/L of glucose released, which represents a novel valorisation of two different side products (NaOH enriched wastewater and cotton textile waste) of the textile industry. The Saccharomyces cerevisiae strain CEN.PK m850, engineered for redirecting the natural alcoholic fermentation towards a homolactic fermentation, was then used to valorise the glucose-enriched hydrolysate into lactic acid. Overall, the process produced 53.04 g/L ± 0.34 of l-lactic acid, with a yield of 82.7%, being the first example of second-generation biomass valorised with this yeast strain, to the best of our knowledge. Remarkably, the fermentation performances were comparable with the ones obtained in the control medium.ConclusionThis study validates the exploitation of cotton post–industrial waste as a possible feedstock for the production of commodity chemicals in microbial cell-based biorefineries. The presented strategy demonstrates the possibility of implementing a circular bioeconomy approach in manufacturing textile industries.Graphical