Yeast Fermentation Research Articles

Utilization of cotton stalk waste for sustainable isopropanol production via hydrolysis and coculture fermentation

The present investigation focused on isopropanol production from lignocellulosic cotton stalk biomass (CSB) using conventional pretreatment, enzymatic hydrolysis and fermentation methods. In comparison to alkaline (NaOH) pretreatment, dilute sulphuric acid (H2SO4) pretreatment showed higher cellulose exposure (448.5 mg/g). Further, ultrasono assisted acid and alkali pretreatment was performed for maximum exposure of cellulose and it was found 616.9 and 586.15 mg/g respectively. Chemical pretreated CSB was additionally exposed to independent enzymatic hydrolysis using commercial enzymes (Celluclast and Viscozymes) following Response Surface Methodology (RSM), which revealed a maximal production of glucose and xylose yield (544.6 mg/g and 41.8 mg/g). Pretreated and enzymatic hydrolyzed cotton stalk biomass at various conditions were analysed using SEM, FTIR, and XRD to determine the structural and functional changes. Further, a co-culture strategy was employed on pretreated and hydrolyzed CSB using two fermented yeast strains (Saccharomyces cerevisiae and Pichia pastoris) for isopropanol production. HR-MS analysis revealed that the maximum concentration of isopropanol (126.228 mM) was produced in 2:1 proportionate ratio of two fermented yeasts with 20 g/L of substrate loadings at 72 h of incubation time. These results indicate that the production of isopropanol (7.46 g/L) from CSB with different parametric conditions is an encouraging step and can be exploited further for various industrial applications.

Effect of Sourdough-Yeast Co-Fermentation on Physicochemical Properties of Corn Fagao Batter.

Fagao is one of China's traditional gluten-free staple foods made with rice or corn flour. Corn Fagao prepared by co-fermentation with sourdough and yeast exhibits better quality and less staling compared to traditional yeast-fermented Fagao. The physicochemical properties of corn Fagao batter during sourdough-yeast co-fermentation were investigated. The results showed that compared with yeast fermentation, the gas production and viscosity of the batter increased with co-fermentation. The co-fermented batter showed a higher hydrolysis of starch and less amylose content. The integrity of starch granules in the co-fermented batter was damaged more seriously, and the crystallinity and short-range ordered structure were less than in the yeast-fermented batter, even though the crystal structure type of starch did not obviously change. The peak viscosity, minimum viscosity, final viscosity, decay value, and recovery value of the corn batter were reduced by co-fermentation, which improved the thermal stability of the batter and slowed down the aging. Co-fermentation also resulted in a more pronounced reduction in protein subunit content than yeast fermentation. The changes in the physicochemical properties of the corn Fagao batter help explain the improvement in quality of corn Fagao made from the co-fermentation method and may provide theoretical references for co-fermentation with sourdough and yeast to other gluten-free foods.

Metabolization of Free and Peptide-Bound Oxidized Methionine Derivatives by Saccharomyces cerevisiae in a Model System.

In the brewing process, methionine is a decisive amino acid for (off-)flavor formation. A significant part of methionine is oxidized to methionine sulfoxide (MetSO) in malt. We hypothesized that MetSO and MetSO2 are metabolized to volatile compounds during yeast fermentation and examined whether the yeast Saccharomyces cerevisiae is able to catabolize l-MetSO and l-MetSO2 in free and dipeptide-bound forms. We also investigated the stability of l-methionine sulfoximine and S-methylmethionine. Cell viability in the presence of the test compounds was at least 90%. Both free and peptide-bound test substances were metabolized by Saccharomyces cerevisiae. l-MetSO was degraded most rapidly as the free amino acid, while l-MetSO2 was degraded most rapidly bound in dipeptides. We observed a different degradation behavior of the (R) and (S) diastereoisomers for l-MetSO and l-methionine sulfoximine. Furthermore, we detected methionol as the only metabolite of MetSO. Methionol sulfoxide was not formed. MetSO2 was not converted to methionol or methionol sulfone but to the respective α-hydroxy acid. We conclude that the reduction of MetSO to methionine proceeds faster than transamination. The occurrence of MetSO or MetSO2 in brewing malt will not lead to the formation of hitherto unknown volatile metabolites of the Ehrlich pathway.

Role of cultivation parameters in carbohydrate accretion for production of bioethanol and C-phycocyanin from a marine cyanobacterium Leptolyngbya valderiana BDU 41001: A sustainable approach

The investigation aimed to augment carbohydrate accumulation in the marine cyanobacterium Leptolyngbya valderiana BDU 41001 to facilitate bioethanol production. Under the standardised physiochemical condition (SPC), i.e. 90 µmol photon m−2 s−1 light intensity, initial culture pH 8.5, 35 °C temperature and mixing at 150 rpm increased the carbohydrate productivity ∼70 % than the control, while a 47 % rise in content was obtained under the nitrate (N)-starved condition. Therefore, a two-stage cultivation strategy was implemented, combining SPC at the 1st stage and N starvation at the 2nd stage, resulting in 80 % augmentation of carbohydrate yield, which enhanced the bioethanol yield by ∼86 % as compared to the control employing immobilised yeast fermentation. Moreover, biomass utilisation was maximised by extracting C-phycocyanin, where a ∼77 % rise in productivity was recorded under the SPC. This study highlights the potential of L. valderiana for pilot-scale biorefinery applications, advancing the understanding of sustainable biofuel production.

Effects of cereal flour types and sourdough on dough physicochemical properties and steamed bread quality.

Coarse cereals have been promoted for their health benefits, and sourdough is used to improve their steamed bread sensory acceptance. However, grains vary in dough physiochemical properties and steamed bread-making performance. This study investigated the effects of yeast and sourdough fermentation on the biochemical, textural, and flavor properties of dough and steamed bread of eight grain types. Results indicated that sourdough dough had a lower pH and higher total titrable acidity compared with yeast group. The texture of sourdough-steamed bread was significantly improved with reduced hardness and enhanced springiness. Microstructure revealed that sourdough resulted in starch surface corrosion and less amylopectin recrystallization. Aldehydes, alcohols, and esters are more dominant in sourdough group than yeast group. Foxtail millet and sorghum steamed breads exhibited the highest performances in texture, flavor, and sensory evaluation. This could provide a theoretical basis for producing coarse cereal products with desirable quality.

Effects of unconventional non-Saccharomyces yeast fermentation on the chemical profile and bioaccessibility of watermelon wine

This study evaluated effects of watermelon fermentation using different non-Saccharomyces yeast strains (Brettanomyces lambicus, Brettanomyces bruxellensis, and Lachancea thermotolerans) on the chemical profile of the derived wines. A control product was produced using Saccharomyces cerevisiae. Analytical techniques, namely HPLC-DAD-RID and LLE-GC/MS, were used to evaluate sugars, organic acids, alcohols, phenolic compounds, amino acids, and volatile compounds profiles. Fermentation reduced the sugar (glucose and fructose) and amino acid contents and increased alcohol (ethanol and glycerol), organic acids (lactic and acetic acids), phenolics, and volatile compounds. The use of the B. bruxellensis strain resulted in an alcoholic beverage with quality parameters similar to that from S. cerevisiae but with higher concentrations of phenolics (rutin, gallic acid, caftaric acid, and caffeic acid) and important volatile compounds (isoamyl acetate). The watermelon wine had a higher content of bioaccessible phenolic compounds than the must, especially catechin, procyanidin B1, procyanidin B2 and gallic acid, due to their high bioaccessibility (>70%). The yeast type influenced the chemical profile of the beverages. B. bruxellensis is a promising tool to obtain watermelon alcoholic beverage in an industrial scale.

Assessment of dietary yeast-based additives for cultured catfish and tilapia health.

Channel catfish (Ictalurus punctatus) and Nile tilapia (Oreochromis niloticus) are two aquaculture species of great importance. Intensive production is often hindered by poor growth performance and disease mortality. The aim of this study was to evaluate the potential of a commercial fermented yeast product, DVAQUA, on channel catfish and Nile tilapia growth performance metrics and disease resistance. Channel catfish and Nile tilapia were fed practical diets supplemented with 0%, 0.1% or 0.4% of DVAQUA over approximately 2-month feeding periods in recirculation aquaculture systems. To assess the potential of the postbiotic against common aquaculture pathogens, juvenile catfish were subsequently challenged by immersion with Edwardsiella ictaluri S97-773 or virulent Aeromonas hydrophila ML09-119. Nile tilapia juveniles were challenged by injection with Streptococcus iniae ARS-98-60. Serum lysozyme activity, blood chemistry and growth metrics were measured at the end of the feeding period, but no differences were observed across the different metrics, except for survival. For the pathogen challenges, there were no differences in endpoint mortality for channel catfish with either pathogen (p > .05). In contrast, Nile tilapia survivability to S. iniae infection increased proportionally to the inclusion of DVAQUA (p = .005). Changes to sera lysozyme activity were also noted in the tilapia trial, with a reduction of activity in the fish fed the 0.4% DVAQUA diet compared to the control diet (p = .031). Expression profiles of proinflammatory genes and antibodies were also found to be modulated in channel catfish fed the postbiotic, indicating some degree of protective response. These results suggest that this postbiotic may be beneficial in protecting Nile tilapia against S. iniae infection by influencing immune parameters and additional research is needed to evaluate the potential of this DVAQUA for improving catfish health and disease control.

Different Nitrogen Consumption Patterns in Low Temperature Fermentations in the Wine Yeast Saccharomyces cerevisiae.

Nowadays, the wine industry carries out fermentations at low temperatures because this oenological practice clearly improves the aromatic complexity of the final wines. In addition, nitrogen content of the must also influences the quality of the wine. In this study, we carried out a phenotypic and fermentative analysis of two industrial wine Saccharomyces cerevisiae strains (P5 and P24) at 15 and 28 °C and three nitrogen concentrations (60, 140 and 300 mg N/L) in synthetic must. Our results show that both parameters, temperature and nitrogen, are interrelated and clearly determine the competitiveness of the wine strains and their ability to adapt at low temperatures. The best adapted strain at low temperatures decreased its competitiveness at lower nitrogen concentrations. In addition, our results show that it is not only the quantity of nitrogen transported that is important but also the quality of the nitrogen source used for wine yeast adaptation at low temperatures. The presence of some amino acids, such as arginine, branched chain amino acids, and some aromatic amino acids can improve the growth and fermentation activity of wine yeasts at low temperatures. These results allow us to better understand the basis of wine yeast adaptation to fermentation conditions, providing important information for winemakers to help them select the most appropriate yeast strain, thus reducing the economic costs associated with long and sluggish fermentations. The correlation between some amino acids and better yeast fermentation performance could be used in the future to design inactive dry yeast enriched in some of these amino acids, which could be added as a nutritional supplement during low temperature fermentations.

Brewing with Sea Vegetable: The Effect of Spirulina (Arthrospira platensis) Supplementation on Brewing Fermentation Kinetics, Yeast Behavior, and the Physiochemical Properties of the Product

Spirulina is a highly nutritious microalgae commonly used as a food additive. During fermentation, different adjuncts are incorporated to act as a nutrient source for yeast and fortify or modify the sensory attributes of the final product. In this study, the effect of Spirulina on the characteristics of controlled yeast fermentation and the production of volatile organic compounds (VOCs) was analyzed. Spirulina was added to malted barley during mashing and fermented under standard conditions. An unaltered mash (negative control) and yeast extract (positive control) were also fermented. The addition of Spirulina resulted in an increased fermentation rate (~14% faster) and bigger yeast cells (~34% larger) in comparison to the negative control. There were differences in color (determined as SRM) between treatments; however, there were only minor differences in VOCs, with no statistical differences observed between chemical compound groups. No differences were observed in the pH, total number of yeast cells, or final attenuation between treatments. The primary mechanism for the observed differences is believed to be an increase in amino acids available to yeast that were contributed by the Spirulina. This shows both that Spirulina has a high potential as a fermentation adjunct and that the amino acid profile of an adjunct can significantly impact fermentation.

Nitrogen Availability and Utilisation of Oligopeptides by Yeast in Industrial Scotch Grain Whisky Fermentation

Scotch grain whisky is produced with a substantial proportion of unmalted grains, which can result in nitrogen deficiency for yeast in the fermentable grain mash. This study examined nitrogen source availability and utilisation by three commercial whisky strains (Saccharomyces cerevisiae) during Scotch grain whisky fermentation, focusing on oligopeptides. Peptide uptake kinetics in synthetic whisky mash with defined peptides showed that oligopeptides of up to nine amino acids were taken up by the strains, albeit with some variability between the strains. The study found that peptides with appropriate molecular weights could replace free amino acids without negatively affecting fermentation kinetics. Moreover, fermentation performance improved when additional nitrogen was provided via peptides rather than diammonium phosphate. Analysis of industrial grain mash indicated that despite low initial yeast assimilable nitrogen, residual proteolytic activity from malt increased nitrogen availability during fermentation. Approximately 30% of the nitrogen consumed by yeast during grain mash fermentation was derived from peptides. LC-HRMS peptide analysis revealed complex dynamics of peptide formation, degradation, and utilisation. This study highlights the importance of oligopeptides in ensuring optimal fermentation efficiency in Scotch grain mash and similar substrates.

Biological and chemical enhancements of Elaeocarpus sylvestris var. ellipticus through fermentation: Implications for therapeutic and industrial applications

This study aimed to evaluate the biological and chemical improvements that take place during the yeast fermentation of hot water-extracted Elaeocarpus sylvestris (HES). HES was diluted with sterile distilled water to a final concentration of 1:200 (v/v), followed by the inoculation of Pichia fermentans (PF) and Saccharomyces cerevisiae (SC) were inoculated as starters. The fermentation by yeast strains proceeded for five days, during which the viable cell count and pH were monitored. The radical scavenging activity was assessed using DPPH and ABTS assays. The impact of HES on proinflammatory cytokines in RAW 264.7 cells and cytotoxicity in HT-29, MCF-7, and INT407 cells was examined. Changes in metabolic profile during fermentation were analyzed using GC-TOF-MS and UPLC-Orbitrap-MS/MS. HES fermented with PF and SC maintained viable cell counts throughout the 5-day fermentation period. The pH levels were consistently within the range of 3.62 ± 0.03 to 3.65 ± 0.15. The expression of TNF-α and IL-6 significantly decreased at all concentrations (31–500 mg/mL) in RAW cells treated with lipopolysaccharide and HES, with cytokine expression levels returning to baseline levels. The cell viability of INT407 significantly increased following HES treatment (p < 0.05). Fermentation of HES with PF or SC led to significant changes in the composition ratios of carboxylic acids and derivatives, and organooxygen compounds. The HES extract demonstrates potential applicability as a raw material with antioxidative, anti-inflammatory, and anticancer properties for applications in the food and cosmetic industries.

Effect of yeast (Saccharomyces cerevisiae) fermentation on conformational changes in pig liver proteins and their ability to bind to characteristic aldehydes

This study aimed to explore the potential of a fermentation technology to reduce off-flavour perception and its underlying mechanisms. Results revealed that yeast fermentation (YF) significantly ameliorated the off-flavour of pig liver (p < 0.05). Specifically, YF pre-treatment decreased the relative abundance of α-helix and fluorescence intensity while increasing the surface hydrophobicity and SS level and loosening the microstructure of myofibrillar proteins (MPs) in pig liver. Additionally, the appropriate fermentation treatments enhanced the MP–aldehyde binding capacity by 0.25–1.30 times, demonstrating that YF-induced conformational modifications in pig liver proteins made them more prone to interacting with characteristic aldehydes. Moreover, molecular docking results confirmed that hydrophobic interactions are the primary drivers of MP–aldehyde binding. These findings suggest that YF technology holds immense promise for modulating off-flavour perception in liver products by altering protein conformation.

Engineered Chlorella vulgaris improves bioethanol production and promises prebiotic application.

Microalgal biomass for biofuel production, integration into functional food, and feed supplementation has generated substantial interest worldwide due to its high growth rate, non-competitiveness for agronomic land, ease of cultivation in containments, and presence of several bioactive molecules. In this study, genetic engineering tools were employed to develop transgenic lines of freshwater microalga Chlorella vulgaris with a higher starch content, by up-regulating ADP-glucose pyrophosphorylase (AGPase), which is a rate-limiting enzyme in starch biosynthesis. Expression of the Escherichia coli glgC (AGPase homolog) gene in C. vulgaris led to an increase in total carbohydrate content up to 45.1% (dry cell weight, DCW) in the transgenic line as compared to 34.2% (DCW) in the untransformed control. The starch content improved up to 16% (DCW) in the transgenic alga compared to 10% (DCW) in the control. However, the content of total lipid, carotenoid, and chlorophyll decreased differentially in the transgenic lines. The carbohydrate-rich biomass from the transgenic algal line was used to produce bioethanol via yeast fermentation, which resulted in a higher ethanol yield of 82.82mg/L as compared to 54.41mg/L from the untransformed control. The in vitro digestibility of the transgenic algal starch revealed a resistant starch content of up to 7% of total starch. Faster growth of four probiotic bacterial species along with a lowering of the pH of the growth medium indicated transgenic alga to exert a positive prebiotic effect. Taken together, the study documents the utilization of genetically engineered C. vulgaris with enriched carbohydrates as bioethanol feedstock and functional food ingredients.

Valorization of process wastes from soft-wheat processing industry through biofuel production.

A substantial amount of process waste is generated during the manufacture of soft-wheat products (SWPs), such as biscuits/cookies, crackers, wafers, and cakes. A small portion of waste is reused in specific biscuits, whereas the rest is usually discarded. This study aimed to investigate the suitability of this waste for the co-production of bioethanol and fatty acid methyl esters (FAMEs or biodiesel). Two groups of waste generated in the SWP industry were included in the study: (a) the waste of low-moisture (<10%) biscuits, crackers, and wafer sheets with no fillings and/or coatings, and (b) the waste of high-moisture (>10%) biscuits, crackers, wafers, and cakes with fillings and/or coatings. The study involved extracting each sample with hexane, and the recovered fat was converted to the FAME through alkali-catalyzed transesterification. The remaining carbohydrate-rich fraction was then converted to bioethanol through amylolytic hydrolysis and yeast fermentation. A great portion (92.42%-93.17%) of the fat was extracted from the wastes and converted to the FAME with adequate yields (13.81-14.55g FAME/g waste, dm) and acceptable conversion efficiencies (85.19%-89.04%). However, bioethanol production from the defatted carbohydrate-rich fractions proceeded rather slowly, yielding only 16.54-18.02 (g ethanol per g of waste, dm), corresponding to fermentation efficiencies ranging from 43.32% to 48.29%. Upon the co-production of FAME and ethanol, a considerable amount (50.93%-53.08%) of waste solids remained in the residue fraction. These findings indicated that production of the FAME with adequate yields and conversion efficiencies is viable from the SWP industry wastes; however, bioethanol yields and fermentation efficiencies are rather limited, which warrants further investigation. PRACTICAL APPLICATION: The soft-wheat processing industry generates 1%-5% of total production as waste. The waste was studied to produce FAME and bioethanol. The fat was extracted from the waste and converted to FAME. Bioethanol yields and fermentation efficiencies are limited due to dough modifiers and antimicrobial additives used in SWP production. Further research is required to improve ethanol yield.

Cultivating black soldier fly (Hermetia illucens) larvae on coconut endosperm and soybean curd residue: Impact on growth performance, waste reduction efficiency and larval nutritional composition

AbstractBlack soldier fly larva (BSFL), Hermetia illucens (L.) (Diptera: Stratiomyidae), presents an attractive waste management solution that not only addresses the issue of waste but also generates alternative proteins and fats. Substrate quality and pre‐treatment by fermentation may play a crucial role in waste reduction efficiency and larval quality. The aim of this study was to investigate the growth performance, waste reduction efficiency and nutritional composition of BSFL rearing using different ratios of coconut endosperm (CE) and soybean curd residue (SC) with pre‐treatment by self‐fermentation (F), fermentation with baker's yeast (Y) and without pre‐treatment (C). Seven‐day‐old larvae were randomly separated into 15 experimental groups with three replicates. The experiment ended when the prepupae appeared. Based on the results, the larvae fed a diet with a high percentage of fresh SC exhibited the highest weight and growth rate, whereas the shortest development time was observed in larvae reared on Y. Pre‐treatment of the substrate with fermentation proved effective in waste reduction. The nutritional composition of the larvae showed that the highest crude protein was produced in larva reared on fresh 100% SC, and the highest ether extract was found when fed CE at 100% with pre‐treatment with F or Y. In conclusion, it is recommended to employ a fresh diet with a high SC content to optimise protein production, whereas a high level of CE with F or Y may be performed to obtain a high ether extract content. However, incorporating yeast fermentation with an SC content exceeding 25% provided high efficiency in waste reduction. Therefore, the selection of the ratio between CE and SC and of pre‐treatment techniques depends on the producer's objective.

A MATLAB-based simulator for the study of process control of fed-batch yeast fermentations

This paper presents a MATLAB-based simulator for the study of process control of fed-batch yeast fermentations that meets the educational needs of undergraduate and graduate students. Against the background of challenges in interdisciplinary education, the paper examines the evolving environment of simulation tools in the field of bioprocesses. It emphasizes the need for interdisciplinarity to prepare students for the complexities of the modern biotechnology industry. Built with accessibility and flexibility in mind, the simulator offers a modular structure with a graphical user interface (GUI) for novice users and direct access to MATLAB functions for advanced users. This design choice ensures ease of use for students with different programming backgrounds and allows for adaptability to alternative software environments. The introductory sections provide a detailed overview of the simulator development, including the mathematical models that govern biomass growth kinetics, mass transfer, and process quality indicators. Selected process control strategies such as rule-based and feedforward approaches are incorporated in the simulator to enhance the learning experience by allowing students to experiment with different scenarios. The paper concludes by emphasizing the simulator's adaptability, modularity, and user-friendly interface as a valuable asset in educating students about the complexities of bioprocess control.

Critique: YEASTsim - A Matlab-based simulator for teaching process control in fed-batch yeast fermentations

One of the primary challenges in chemical engineering education lies in the practical application of theoretical knowledge. Contemporary society demands chemical engineers who are not only adept at critical thinking but also proficient in practical problem-solving. Historically, acquiring such practical experience has been heavily dependent on laboratory experiments. However, with the advent of advanced technology, there are now more accessible and varied opportunities for experiential learning. Simulation-based learning tools have proven to be powerful complements to traditional laboratory experiments. These tools enable students to investigate complex chemical processes, test hypotheses, and develop practical skills in a safe, cost-effective, and scalable environment. This critique aims to evaluate the educational potential of the YEASTsim simulator, positioning it as an interactive and innovative tool to enhance the study of core subjects such as Chemical Reaction Engineering, a fundamental component of chemical engineering practice.

Extraction of polyphenols and antioxidant compounds from SCOBY, as a by-product of Kombucha, using different types of extraction

Kombucha is obtained through a symbiosis fermentation of bacteria and yeast, generating a floating film called SCOBY (Symbiotic Culture of Bacteria and Yeast). After a period of use, SCOBY becomes a production waste. Thus, alternative uses of SCOBY can add value to Kombucha production. In this way, this work aimed to produce and evaluate antioxidant activity and total phenolic contents of SCOBY ethanolic extracts, which were obtained by two different solid–liquid extraction with three different waste weight/extractor solvent volume ratios. The SCOBY evaluated had a high concentration of cellulose (9.42 ± 1.00%) and low concentrations of proteins (0.84 ± 0.04%) and lipids (0.29 ± 0.05%). All evaluated extracts showed a high concentration of total phenolics (40.7–64.3 mg GAE/100 g of SCOBY) and high antioxidant activity (ABTS•+, FRAP and ORAC assays). The most efficient extraction of bioactive compounds was obtained by decoction using the highest solvent volume (ratio 1:20). The PCA showed clusters of the samples both by the type of extraction and by the different ratios of the weight of SCOBY to the solvent volume. Of our knowledge, our work is the first in the literature that describes such data. Therefore, the present work showed the possibility of using this industrial waste mainly to obtain active compounds and antioxidants.

RETRACTED: Wong et al. In-Situ Yeast Fermentation Medium in Fortifying Protein and Lipid Accumulations in the Harvested Larval Biomass of Black Soldier Fly. Processes 2020, 8, 337

The Processes Editorial Office retracts the article titled “RETRACTED: In-Situ Yeast Fermentation Medium in Fortifying Protein and Lipid Accumulations in the Harvested Larval Biomass of Black Soldier Fly” [...]

Use of non-&lt;i&gt;Saccharomyces&lt;/i&gt; yeasts in the &lt;i&gt;pris de mousse&lt;/i&gt; of Lambrusco. Microbial evolution through alcoholic fermentation and effect on wine volatile profile

Lambrusco is a sparkling wine largely produced in the north of Italy, especially in the Emilia region. The possible application of two non-Saccharomyces yeasts in the secondary bottle fermentation (Champenoise method) was tested in combination with a commercial strain of S. cerevisiae to obtain wines having a distinctive volatile profile. Our results demonstrated that the gradual increase of ethanol content in the pied de cuve ensured the adaptation of Hanseniaspora guilliermondii and Torulaspora delbrueckii at the bottle fermentation, with survival comparable with that of S. cerevisiae, in the order of 6 log units. The simultaneous presence of two yeast species reduced the maximum fermentation rate, without any relevant alteration in the main oenological parameters of resultant wines. GC MS-MS analysis of the volatile profile of wines (46 compounds) highlighted differences in wine made from a pure culture of S. cerevisiae from wines obtained by mixed yeasts. Acetates, esters, and fatty acids are the classes of volatile compounds mostly affected by using different yeasts in the bottle fermentation of Lambrusco wines. This work provided for the first time information about the volatile profile of Lambrusco and suggests an innovative application of non-Saccharomyces yeast in the production of sparkling wines by champenoise methods.