Yeast Viability Research Articles

Chemical and microbiological assessment of early wine fermentation phase can predict yeast cell viability during post-fermentation process.

The management of post-fermentation phase is essential for the protection of wine oxidation. The prolonged contact of yeast lees and wine can help to limit this problem, although off-flavours can originate. It is known that some cellular components (mannoproteins, lipids, glutathione, etc.) released into the wine influence oxygen protection; however, still active cells could contribute to maintaining protection against oxidation. To date, in the literature there is a lack of data that evaluates cell viability, especially in the post-fermentation phase, particularly using methods different by plate count that identifies only a small part of the viable population. The aim of the work was to investigate the yeast viability of 12 wine Saccharomyces cerevisiae strains during 45days after the fermentation in natural grape juice. The major fermentation parameters were measured at early phase (40h) and at the end of the process, and were correlated with total and viable cells in the post-fermentation phase. Contrary to what has been observed in the literature, this work demonstrates that cell viability in the post-fermentation phase is very high and dependent on the yeast strain. A predictive model that can estimate viability in the post-fermentation phase, based on parameters measured at the early fermentation phase, was successfully set up. This approach can be adopted by wineries or winemakers as it uses fermentation data (sugar and nitrogen residues, ethanol and glycerol production, total cell count) obtained through simple chemical and microbiological analyses.

UV-C and Nanomaterial-Based Approaches for Sulfite-Free Wine Preservation: Effects on Polyphenol Profile and Microbiological Quality.

Controlling the microorganisms employed in vinification is a critical factor for successful wine production. Novel methods aimed at lowering sulfites used for wine stabilization are sought. UV-C irradiation has been proposed as an alternative for reducing the viable cell count of microorganisms in wine and grape juice. Nevertheless, UV-C treatment poses the risk of altering the chemical properties of wine. Therefore, this study aimed to test and implement iron oxide-silica core-shell nanomaterial functionalized with TiO2 in UV-C treatment of white and red wines. Material for the study consisted of the synthesized nanocomposite, Saccharomyces cerevisiae as a model yeast, and Muscaris and Cabernet Cortis wines. The viability of yeasts under treatment, the physiochemical properties of wine, and polyphenol content were tested. Studies have shown that nanomaterial can modulate the effects of UV-C treatment regarding yeast viability and polyphenol content, and the effectiveness of the treatment depends on the wine type. These results open up discussion on the possible use of the proposed hurdle technology in winemaking to control the polyphenol composition and alcohol reduction.

The Probiotication of a Lychee Beverage with Saccharomyces boulardii: An Alternative to Dairy-Based Probiotic Products.

Probiotic vegetable-based beverages, such as lychee, can be rich in nutrients, free of cholesterol and lactose, and also have few allergenic components. Saccharomyces boulardii is an alternative to make lychee juice a probiotic beverage. This work aimed to develop probiotic lychee beverage (LB) using S. boulardii by evaluating the effect of refrigerated storage on cell viability, physicochemical characteristics, and acceptance. LB supplemented with S. boulardii was fermented (24 h/30 °C), supplemented with sucrose (4-12 °Brix), and refrigerated (up to 28 days/4 °C). The yeast viability, total soluble solid (TSS) and, ethanol content, pH, total phenolic compounds (TPC), and antioxidant activity (AA) levels were evaluated over 28 days of storage. Also, the profiles of sugars, organic acids, and phenolic were determined via chromatographic analysis. The sensory acceptance of the probiotic beverage was evaluated. Higher sucrose levels (12 °Brix) resulted in greater yeast viability (6.9 log CFU/mL) on the 21st day of storage and superior TPC (153 µmol TEAC/mL) and ethanol levels (8.7%). Storage reduced the TPC, AA, and TSS. LB supplemented with sucrose to 12 °Brix, probioticated by S. boulardii, and stored for 21 days became accepted by the consumer and presented an adequate physicochemical profile with probiotic potential. The probiotication of lychee beverage is an alternative to dairy-based probiotic beverages.

The protective effect of trehalose and monosodium glutamate on yeast viability and antagonistic properties during freeze-drying

The protective effect of trehalose and monosodium glutamate on yeast viability and antagonistic properties during freeze-drying

Development and characterization of microencapsulated Pichia kluyveri CCMA 0615 with probiotic properties and its application in fermented beverages

The study aimed to develop innovative microencapsulated formulations of strains with probiotic attributes, Pichia kluyveri CCMA 0615 and Saccharomyces cerevisiae CCMA 0732. The yeasts (8 log CFU/mL) were microencapsulated by spray drying technique using whey powder (WP - 15 %, 20 %, and 30 %) and sodium alginate (ALG - 1 %). The microcapsules and cell viability were characterized during two months of storage (4 °C and 25 °C). The selected formulations were applied to functional beverage fermentation, and viability and survival in the simulated gastrointestinal tract (GIT) were performed. The viability of yeasts microencapsulated by the spray drying method was shown to be dependent on the strain and encapsulating matrix used, ranging from 84 to 99 %. P. kluyveri required refrigeration when storing microcapsules. In functional beverage fermentation, microencapsulated yeast maintained the same fermentative profile with carbohydrate consumption, production of lactic acid (0.30 to 1.10 g/L) and alcohol (0.2 to 1.61 g/L), and greater viability during storage. Finally, the microencapsulation of P. kluyveri with 15 % WP + 1 % ALG maintained high viability under GIT conditions, whether exposed independently (>84 %) or incorporated into a food matrix (>94 %). The study demonstrated that this innovative microencapsulation of probiotic yeasts increases their viability, improves biotechnological application, and facilitates efficient delivery of probiotics to the host.

Photodynamic therapy reduces viability, enhances itraconazole activity, and impairs mitochondrial physiology of Sporothrix brasiliensis

Sporothrix brasiliensis is the main agent of sporotrichosis in Brazil, with few therapeutic options. This study aimed to investigate the in vitro efficacy of photodynamic therapy using a diode laser (InGaAIP) in combination with the photosensitizer methylene blue against S. brasiliensis yeasts. Additionally, we evaluated the underexplored mitochondrial activity of S. brasiliensis and the impact of laser treatment on the fungal mitochondrial aspects post-treatment. Three strains of S. brasiliensis were used, including a non-wild-type strain to itraconazole. Yeast viability was determined by counting colony-forming units. For a comprehensive analysis of irradiated versus non-irradiated cells, we assessed combined therapy with itraconazole, scanning electron microscopy of cells, and mitochondrial activity. The latter included high-resolution respirometry, membrane potential analysis, and reactive oxygen species production. Methylene blue combined with photodynamic therapy inhibited the growth of the isolates, including the non-wild-type strain to itraconazole. Photodynamic therapy induced the production of reactive oxygen species, which negatively affected mitochondrial function, resulting in decreased membrane potential and cell death. Photodynamic therapy altered the ultrastructure and mitochondrial physiology of S. brasiliensis, suggesting a new therapeutic approach for sporotrichosis caused by this species.

Supplementation of Chlorella vulgaris Extracts During Brewing: The Effects on Fermentation Properties, Phytochemical Activity and the Abundance of Volatile Organic Compounds

Chlorella vulgaris, a microalga rich in secondary metabolites and nutrients, offers a promising alternative for promoting microbial growth in food fermentation processes. This study aimed to evaluate the effects of C. vulgaris extracts on fermentation kinetics, sensory characteristics, phytochemical composition, antioxidant activity, and the abundance of volatile organic compounds (VOCs) in treated versus control beers. The bioactive compounds from C. vulgaris were extracted using an ultrasound-assisted method with water as the solvent. A German Pilsner-style lager beer (GPB) was brewed and supplemented with 0.5, 1, and 5 g/L of C. vulgaris extracts prior to primary fermentation. Yeast viability, °Brix, and pH levels were monitored to assess fermentation progress. Phytochemical composition was analyzed by quantifying total polyphenols and flavonoids. The antioxidant activity of the beer was evaluated using both the 2,2-diphenyl-1-picrylhydrazyl (DPPH●) and hydrogen peroxide assays. The addition of C. vulgaris extracts resulted in increased yeast viability and slight variations in gravity during the 7-day fermentation period. Moreover, the beers supplemented with C. vulgaris extracts demonstrated higher levels of total polyphenols, flavonoids, and antioxidant activity compared to the GPB. Specific volatile organic compounds, including 2-methyl-1-propanol, 1-hexanol, isopentyl hexanoate, 2-methylpropyl octanoate, β-myrcene, and geranyl acetate, were significantly more abundant (p < 0.05) in the treated beers than in the control. Sensory evaluations revealed a favorable impact of the treatment on aroma scores compared to the GPB. Overall, the findings indicate that C. vulgaris extracts could be a valuable ingredient for developing functional beers with enhanced health benefits, particularly regarding antioxidant activity. Additionally, the results underscore the importance of exploring innovative approaches that utilize natural sources like Chlorella to enrich the nutritional profile and sensory qualities of fermented products.

ML-enhanced peroxisome capacity enables compartmentalization of multienzyme pathway.

Repurposing an organelle for specialized metabolism provides an avenue for fermentable, unicellular organisms such as Saccharomyces cerevisiae to mimic compartmentalization of metabolic pathways within different plant tissues. Peroxisomes are attractive organelles for repurposing as they are not required for yeast viability when grown on glucose and can efficiently compartmentalize heterologous enzymes to enable physical separation of cytosolic native metabolism and peroxisomal engineered metabolism. However, when not required, peroxisomes are repressed, leading to low functional capacities for heterologous proteins. Here we engineer peroxisomes with enhanced functional capacities, with the goal of compartmentalizing up to eight metabolic enzymes to enhance titers. We implement a machine learning pipeline that allows the identification of factors to overexpress, culminating in a 137% increase in peroxisome functional capacity compared to a wild-type strain. Improved pathway compartmentalization enables an 80% increase in the biosynthesis titers of the monoterpene geraniol, up to 9.5 g L-1.

In Vivo Antioxidant Activity of Common Dietary Flavonoids: Insights from the Yeast Model Saccharomyces cerevisiae.

Oxidative stress, associated with diseases and aging, underscores the therapeutic potential of natural antioxidants. Flavonoids, known for scavenging free radicals and modulating cell signaling, offer significant health benefits and contribute to longevity. To explore their in vivo effects, we investigated the antioxidant activity of quercetin, apigenin, luteolin, naringenin, and genistein, using Saccharomyces cerevisiae as a model organism. We performed viability assays to evaluate the effects of these compounds on cell growth, both in the presence and absence of H2O2. Additional assays, including spot assays, drug drop tests, and colony-forming unit assays, were also conducted. Viability assays indicated that the tested compounds are non-toxic. H2O2 reduced yeast viability, but flavonoid-treated cells showed increased resistance, confirming their protective effect. Polyphenols scavenged intracellular reactive oxygen species (ROS) and protected cells from oxidative damage. Investigations into defense systems revealed that H2O2 induced catalase activity and oxidized glutathione accumulation, both of which were reduced by polyphenol treatment. The tested natural compounds enhance cell viability and reduce oxidative damage by scavenging ROS and modulating antioxidant defenses. These results suggest their potential as supplements and pave the way for further research.

Evaluation of different hydration techniques on the viability of commercial dry brewing yeast

The utilization of dry yeast is increasingly becoming established in the brewing context, owing to the ease of storage and handling, the long shelf life and the convenience of dispensing a delicate and costly propagation process for fresh liquid cultures, which would need specialized labor and equipment. Dried yeasts undergo a procedure that removes nearly all liquid contents from the cell, and rehydration is then recommended prior to inoculation into the wort. Hydration is a nuanced procedure that allows yeasts to regain their vitality and viability, however, if carried out improperly, it can adversely affect fermentation. This study aimed to evaluate the impact of four hydration techniques on the viability and vitality of dry commercial yeast cells, preliminarily evaluating the influence of hydration using distilled and filtered water. Two different methodologies were employed, including the assessment of viability, by counting live and dead cells in the Neubauer chamber and the evaluation of vitality, by measuring the CO₂ released after inoculation. The hydration techniques tested included hydration in water without stirring, hydration in water with stirring, and these two techniques replicated in dry malt extract (DME) wort with Specific Gravity (SG) 1040. Firstly, it was found that rehydrating the yeast in distilled water has a negative impact on cell viability. Also, the results indicated that initial agitation has a negative effect on yeast viability and vitality, while hydration in water without agitation produced the best results. Correct hydration of dry yeast is crucial for avoiding off-flavors due to accelerated or delayed fermentation.

The Impact of Cyclodextrins on the Physiology of Candida boidinii: Exploring New Opportunities in the Cyclodextrin Application.

Cyclodextrins, commonly used as excipients in antifungal formulations to improve the physicochemical properties and availability of the host molecules, have not been systematically studied for their effects and bioactivity without a complex active substance. This paper evaluates the effects of various cyclodextrins on the physiology of the test organism Candida boidinii. The research examines their impact on yeast growth, viability, biofilm formation and morphological changes. Native ACD, BCD, randomly methylated α- and β-CD and quaternary ammonium α-CD and β-CD were investigated in the 0.5-12.5 mM concentration range in both static and dynamic systems. The study revealed that certain cyclodextrins exhibited notable antifungal effects (up to ~69%) in dynamic systems; however, the biofilm formation was enhanced in static systems. The magnitude of these effects was influenced by several variables, including the size of the internal cavity, the concentration and structure of the cyclodextrins, and the contact time. Furthermore, the study found that CDs exhibited distinct effects in both static and dynamic systems, potentially related to their tendency to form aggregates. The findings suggest that cyclodextrins may have the potential to act as antifungal agents or growth promoters, depending on their structure and surrounding environments.

Sources of Variability in Determinations of Brewing Yeast Total Cell Counts and Viability Obtained by Microscopy

Brewers utilizing yeast cell counts and determinations of yeast viability obtained by microscopy require precise results for calculating the yeast pitch to produce fermentations with the highest quality and consistency. The steps in obtaining results with the desired precision include identifying, quantifying and controlling the sources of variation. In this study, we used simulated cell counting, manual cell counting, and semi-automated cell counting to explore sources of variation contributing to the variability of yeast cell counts and yeast viability determinations. The results showed that the variability associated with counting different fields of view contributed most to the variability in yeast cell counts and determinations of yeast viability. Understanding the sources and amounts of variation in cell counts will help brewers direct their efforts toward obtaining the highest quality determinations of yeast cell counts and viability.

Design of a Device Based on an LED Matrix for Water Sterilization

The scarcity of potable water emphasizes the urgent need to develop and implement more sustainable treatment technologies, considering both energy consumption and environmental impact. These technologies require effective disinfection systems that avoid the use of chemicals. Innovations in this area, utilizing UV-LED technology, can significantly improve efficiency, reduce costs, and mitigate environmental impacts. This study aimed to evaluate the antimicrobial properties of various encapsulated UV light-emitting diodes (LEDs) to identify the most suitable candidate for constructing an LED array capable of disinfecting large volumes of water. Different devices from various manufacturers, with differing costs and wavelengths, were examined, leading to the selection of the optimal candidate (LED 2) based on its antimicrobial effectiveness and cost-effectiveness. The impact of parameters such as bacterial concentration, sample volume, exposure time, and conditions on disinfection capacity was thoroughly investigated. Exposure to LED 2 resulted in substantial reductions in the viability of bacteria and yeast, demonstrating efficacy even against Clostridium perfringens endospores. Subsequently, an LED array was developed based on these findings and rigorously evaluated for efficacy, confirming its effectiveness as an efficient and environmentally friendly water treatment device.

Molecular characterization of Rft1, an ER membrane protein associated with congenital disorder of glycosylation RFT1-CDG.

The oligosaccharide needed for protein N-glycosylation is assembled on a lipid carrier via a multi-step pathway. Synthesis is initiated on the cytoplasmic face of the endoplasmic reticulum (ER) and completed on the luminal side after transbilayer translocation of a heptasaccharide lipid intermediate. More than 30 Congenital Disorders of Glycosylation (CDGs) are associated with this pathway, including RFT1-CDG which results from defects in the membrane protein Rft1. Rft1 is essential for the viability of yeast and mammalian cells and was proposed as the transporter needed to flip the heptasaccharide lipid intermediate across the ER membrane. However, other studies indicated that Rft1 is not required for heptasaccharide lipid flipping in microsomes or unilamellar vesicles reconstituted with ER membrane proteins, nor is it required for the viability of at least one eukaryote. It is therefore not known what essential role Rft1 plays in N-glycosylation. Here, we present a molecular characterization of human Rft1, using yeast cells as a reporter system. We show that it is a multi-spanning membrane protein located in the ER, with its N and C-termini facing the cytoplasm. It is not N-glycosylated. The majority of RFT1-CDG mutations map to highly conserved regions of the protein. We identify key residues that are important for Rft1's ability to support N-glycosylation and cell viability. Our results provide a necessary platform for future work on this enigmatic protein.

Obtention of pelleted Kluyveromyces marxianus CIDCA 9121 with immunomodulatory properties: Selection of protective agents using Plackett–Burman and Box–Behnken experimental designs

Probiotic yeasts are increasingly used in different animal production systems. Supply of feeding in the form of pellets provides productive advantages in several systems such as poultry production or aquaculture. The inclusion of viable yeast in the pellets has to overcome heat stress during palletization and also dehydration stress. We have worked with our previously characterized probiotic strain Kluyveromyces marxianus CIDCA 9121. We evaluated the resistance of this strain to a process dehydration and extrusion after pellet generation, conditions that were not analyzed for this genus of probiotic yeast. We determined the effects of different thermoprotective agents in order to maximize yeast viability over time and preservation of its probiotic traits. To this aim we used the single factor optimization method, Plackett-Burman central compound rotating design (P–B CCRD) and response surface methodology (RSM) for optimize survival conditions in a process performed at laboratory scale. We selected a combination of sucrose, maltose and maltodextrin and we have tested two different combinations of these agents in a pilot plant extrusion/dehydration process showing that one of them could increase yeast viability. Remarkably, we could observe that immunomodulatory capacity of this strain was not affected after dehydration in the presence of the different additives tested and subsequent rehydration, being the first report showing conservation of probiotic traits of K. marxianus strain upon pelleting opening new perspectives for product development.

Functional analysis of the zinc finger modules of the Saccharomyces cerevisiae splicing factor Luc7.

Identification of splice sites is a critical step in pre-messenger RNA (pre-mRNA) splicing because the definition of the exon/intron boundaries controls what nucleotides are incorporated into mature mRNAs. The intron boundary with the upstream exon is initially identified through interactions with the U1 small nuclear ribonucleoprotein (snRNP). This involves both base-pairing between the U1 snRNA and the pre-mRNA as well as snRNP proteins interacting with the 5' splice site (5'ss)/snRNA duplex. In yeast, this duplex is buttressed by two conserved protein factors, Yhc1 and Luc7. Luc7 has three human paralogs (LUC7L, LUC7L2, and LUC7L3), which play roles in alternative splicing. What domains of these paralogs promote splicing at particular sites is not yet clear. Here, we humanized the zinc finger (ZnF) domains of the yeast Luc7 protein in order to understand their roles in splice site selection using reporter assays, transcriptome analysis, and genetic interactions. Although we were unable to determine a function for the first ZnF domain, humanization of the second ZnF domain to mirror that found in LUC7L or LUC7L2 resulted in altered usage of nonconsensus 5'ss. In contrast, the corresponding ZnF domain of LUC7L3 could not support yeast viability. Further, humanization of Luc7 can suppress mutation of the ATPase Prp28, which is involved in U1 release and exchange for U6 at the 5'ss. Our work reveals a role for the second ZnF of Luc7 in splice site selection and suggests that different ZnF domains may have different ATPase requirements for release by Prp28.

Effect of levan polysaccharide on chronological aging in the yeast Saccharomyces cerevisiae

Levan is a fructose-based biopolymer with diverse applications in the medicinal, pharmaceutical, and food industries. However, despite its extensive biological and pharmacological actions, including antioxidant, anti-inflammatory, and antidiabetic properties, research on its anti-aging potential is limited. This study explored levan's impact on the chronological lifespan (CLS) of yeast Saccharomyces cerevisiae for the first time. The results show that levan treatment significantly extended the CLS of wild-type (WT) yeast by preventing the accumulation of oxidative stress markers (reactive oxygen species, malondialdehyde, and protein carbonyl content) and ameliorating apoptotic features such as reduced mitochondrial membrane potential, loss of plasma membrane integrity, and externalization of phosphatidylserine. By day 40 of the CLS, a significant increase in yeast viability of 6.8 % (p < 0.01), 11.9 % (p < 0.01), and 20.8 % (p < 0.01) was observed at 0.25, 0.5, and 1 mg/mL of levan concentrations, respectively, compared to control (0 %). This study's results indicate that levan treatment substantially modulates the expression of genes involved in the TORC1/Sch9 pathway. Moreover, levan treatment significantly extended the CLS of yeast antioxidant-deficient mutant sod2Δ and antiapoptotic gene-deficient mutant pep4Δ. Levan also extended the CLS of signaling pathway gene-deficient mutants such as pkh2Δ, rim15Δ, atg1, and ras2Δ, while not affecting the CLS of tor1Δ and sch9Δ.

Characteristics of functional ice cream produced with goat’s milk kefir in combination with mono-acylglycerol

&lt;p class="MDPI17abstract"&gt;&lt;strong&gt;Objective:&lt;/strong&gt; The purpose of this study was to determine the character of ice cream made from goat’s milk kefir (GMK) combined with mono-acylglycerol (MAG).&lt;/p&gt;&lt;p class="MDPI17abstract"&gt;&lt;strong&gt;Methods:&lt;/strong&gt; GMK was produced by pasteurized goat’s milk (85&lt;sup&gt;o&lt;/sup&gt;C for 15 s) and 2% kefir grain. Ice cream mix (ICM) was produced by GMK, MAG (T0= 0%, T1= 0.25%; T2= 0.50%; T3= 0.75%), 4% sucrose, and 0.1% salt. The results of observations processed using Analysis of Variance (ANOVA) and Duncan’s Multiple Range Test (DMRT) if there were a significant difference between treatments.&lt;/p&gt;&lt;p class="MDPI17abstract"&gt;&lt;strong&gt;Results:&lt;/strong&gt; Es puter (EP) made from GMK in combination with MAG has a distinctive character against T0 (control). The use of GMK decreases the pH value, thereby increasing the lactic acid and ethanol content which brings out the acidity of the product (p&amp;lt;0.05). In addition, increasing MAG concentration can improve the EP’s overrun to make the product more melt resistant (p&amp;lt;0.05). The use of GMK combined with MAG can support the viability of lactic acid bacteria (LAB) and yeast through storage in the freezer. The number of viable LAB and yeast meets WHO standards to obtained health benefit from consuming probiotic ice cream.&lt;/p&gt;&lt;p class="MDPI17abstract"&gt;&lt;strong&gt;Conclusions:&lt;/strong&gt; To conclude, EP made from GMK in combination with 0.50% MAG was proved to be the best treatment.&lt;/p&gt;&lt;p&gt; &lt;/p&gt;

Evaluation of refractance window-dried type 3 sourdough as an alternative to liquid sourdough in bread production

The aim of this study was to produce type 3 sourdough using refractance window drying (RWD) and to compare the effects of dried and liquid sourdough on bread quality. The drying experiments resulted in a similar viability loss in the total LAB counts of type 3 sourdough samples (p < 0.05), with a 3-log reduction, and the lowest loss in yeast viability was determined in spontaneous sourdough samples. The specific bread volume was in the range of 3.18–3.24 mL/g in the usage of liquid sourdough and 2.75–3.12 ml/g in type 3 sourdough, respectively. The incorporation of type 3 sourdough increased the hardness of bread. The total cell counts, and area was statistically similar in bread produced using liquid spontaneous sourdough and dried Lb. fermentum. When sensory parameters were evaluated, it became evident that sourdough bread produced by dried sourdough exhibited superior organoleptic qualities when compared to those produced by the liquid ferment. As a conclusion, the findings of this study suggest that the application of RW-dried type 3 sourdough can be considered as an alternative to liquid sourdough, without any adverse effect on the physical and sensory attributes of the sourdough bread.

Application of Proton Ionic Liquid in the Process of Obtaining Bioethanol from Hemp Stalks

In this work, hemp (Cannabis sativa L.) stalks were pretreated with pyrrolidinium acetate [Pyrr][AC] to increase the availability of cellulose for cellulolytic enzymes and thus improve the production of 2G ethanol from reducing sugars. The process was carried out under different temperature and time conditions: The control sample was raw material and deionized water, and the second sample was kept at 21 °C for 24 h. The third sample was kept at 90 °C for 30 min, and the fourth sample was kept at the same temperature for 24 h. For each pretreatment, the extraction of lignin and hemicellulose was determined, as well as the change in biomass composition before and after pretreatment. The stalks of hemp seed contained 41.54% cellulose, 18.08% lignin and 28. 87% hemicellulose. [Pyrr][AC] used to dissolve seed hemp at 90 °C for 24 h was most effective in the extraction of this biopolymer, extracting 3.1% content. After enzymatic hydrolysis, the highest content of reducing sugars was found for samples of hemp stalks that were first pretreated with proton ionic liquid (PIL) and stirred at 90 °C for 24 h. On the other hand, the highest ethanol content (5.6 g/L) after fermentation and yeast viability (56.7%) after 72 h were obtained in samples pretreated at 90 °C and for 24 h.