Yeast Culture Research Articles

Highly parallelized laboratory evolution of wine yeasts for enhanced metabolic phenotypes

Adaptive Laboratory Evolution (ALE) of microorganisms can improve the efficiency of sustainable industrial processes important to the global economy. However, stochasticity and genetic background effects often lead to suboptimal outcomes during laboratory evolution. Here we report an ALE platform to circumvent these shortcomings through parallelized clonal evolution at an unprecedented scale. Using this platform, we evolved 104 yeast populations in parallel from many strains for eight desired wine fermentation-related traits. Expansions of both ALE replicates and lineage numbers broadened the evolutionary search spectrum leading to improved wine yeasts unencumbered by unwanted side effects. At the genomic level, evolutionary gains in metabolic characteristics often coincided with distinct chromosome amplifications and the emergence of side-effect syndromes that were characteristic of each selection niche. Several high-performing ALE strains exhibited desired wine fermentation kinetics when tested in larger liquid cultures, supporting their suitability for application. More broadly, our high-throughput ALE platform opens opportunities for rapid optimization of microbes which otherwise could take many years to accomplish.

Diversity of Culturable Yeasts in the Feces of Mew Gulls Breeding in Natural and Urban Habitats, with Insights into the Antifungal Susceptibility of the Observed Pathogens

Migratory birds play an important role in the spread of yeasts in the environment over long distances and in different geographical regions. Human activities, in turn, have a major impact on the biology of wild birds and, consequently, on the microbial communities for which birds act as carriers and disseminators. We sought to assess the “response” of the diversity of culturable yeasts in the feces of Mew Gulls to the type of nesting site (natural/anthropogenic) during the breeding season from April to October 2023. We isolated and molecularly identified 26 yeast species. The species composition in the feces of birds from the natural habitat was more diverse, and the diversity increased from April to October. In contrast, the diversity in the feces of birds from the urban habitat decreased from April to October. Analysis of susceptibility to conventional antibiotics (fluconazole, voriconazole, and amphotericin B) using the CLSI BMD (Clinical and Laboratory Standards Institute broth microdilution) method in isolated strains of opportunistic Candida (C. parapsilosis and C. tropicalis) and strains of the emerging pathogen Rhodotorula mucilaginosa showed that the proportion of resistant strains was higher in strains isolated from the feces of birds from the anthropogenic population. Mew Gulls that spent their breeding season near a landfill and flew away for wintering appear to be a source for the spread of pathogenic yeasts with resistance against antifungal agents.

Characterization of SCOBY-fermented kombucha from different regions and its effect on improving blood glucose

Kombucha is a healthy fermented beverage made by fermenting sugar tea with a symbiotic culture of bacteria and yeast (SCOBY). The fermentation performance and antioxidant capacity of kombucha fermented by SCOBY from five different regions were compared in this study. Two kinds of high-quality SCOBY were selected, GKT and XKT. Next, the metabolite profiles of GKT and XKT were analyzed, and 18 and 36 different metabolites were screened from GKT and XKT, respectively. The correlation analysis of the differential metabolites with the antioxidant indexes identified three metabolites with potential hypoglycemic efficacy, 1-O-Sinapoyl-beta-D-glucose, Actinodaphnine, and Biochanin A. Moreover, the potential of GKT and XKT for anti-diabetes was verified by cell experiments. At the same time, the microbiome of GKT and XKT analysis found that Gluconobacter, Komagataeibacter and Pseudomonas were the most common bacteria genera. Differently, the composition of fungal genera in GKT and XKT was differentiated, with the dominant genera being Filobasidium and Starmerella, respectively. Finally, correlation analysis of microbial composition with volatile and functional metabolites showed that Gluconobacter and Komagataeibacter were closely related to flavor substances. In addition, the dominant strain of GKT, Filobasidium, seems to contribute to its good sensory quality, and the dominant strain of XKT, Starmerella, is positively associated with several metabolites with hypoglycemic effects, particularly Biochanin A and Rhoifolin. In this study, XKT was screened with good fermentation performance and strong antioxidant capacity, as well as had anti-diabetic potential. These results provide microbial resources and theoretical support for the development of kombucha starter culture.

Hydrothermal conditioning of oleaginous yeast cells to enable recovery of lipids as potential drop-in fuel precursors

BackgroundLipids produced using oleaginous yeast cells are an emerging feedstock to manufacture commercially valuable oleochemicals ranging from pharmaceuticals to lipid-derived biofuels. Production of biofuels using oleaginous yeast is a multistep procedure that requires yeast cultivation and harvesting, lipid recovery, and conversion of the lipids to biofuels. The quantitative recovery of the total intracellular lipid from the yeast cells is a critical step during the development of a bioprocess. Their rigid cell walls often make them resistant to lysis. The existing methods include mechanical, chemical, biological and thermochemical lysis of yeast cell walls followed by solvent extraction. In this study, an aqueous thermal pretreatment was explored as a method for lysing the cell wall of the oleaginous yeast Rhodotorula toruloides for lipid recovery.ResultsHydrothermal pretreatment for 60 min at 121 °C with a dry cell weight of 7% (w/v) in the yeast slurry led to a recovery of 84.6 ± 3.2% (w/w) of the total lipids when extracted with organic solvents. The conventional sonication and acid-assisted thermal cell lysis led to a lipid recovery yield of 99.8 ± 0.03% (w/w) and 109.5 ± 1.9% (w/w), respectively. The fatty acid profiles of the hydrothermally pretreated cells and freeze-dried control were similar, suggesting that the thermal lysis of the cells did not degrade the lipids.ConclusionThis work demonstrates that hydrothermal pretreatment of yeast cell slurry at 121 °C for 60 min is a robust and sustainable method for cell conditioning to extract intracellular microbial lipids for biofuel production and provides a baseline for further scale-up and process integration.Graphical abstract

Precursors consumption preferences and thiol release capacity of the wine yeasts Saccharomyces cerevisiae, Torulaspora delbrueckii, and Lachancea thermotolerans

The aromatic profile of wine determines its overall final quality, and among the volatile molecules that define it, varietal thiols are responsible for shaping the distinctive character of certain wine varieties. In grape must, these thiols are conjugated to amino acids or small peptides in a non-volatile form. During wine fermentation, yeasts play a principal role in expressing these aromatic compounds as they internalise and cleavage these precursors, releasing the corresponding free and aroma-impacting fraction. Here, we investigate the impact of three wine yeasts (Saccharomyces cerevisiae, Torulaspora delbrueckii and Lachancea thermotolerans) on thiol releasing in synthetic grape must fermentations supplemented with different cysteinylated (Cys-4MSP and Cys-3SH) and glutathionylated (GSH-4MSP and GSH-3SH) precursors. We demonstrate higher consumption levels of cysteinylated precursors, and consequently, higher amounts of thiols are released from them compared to glutathionylated ones. We also report a significant impact of yeast inoculated on the final thiols released. Meanwhile T. delkbrueckii exhibits a great 3SHA releasing capacity, L. thermotolerans stands out because of its high 3SH release. We also highlight the synergic effect of the co-inoculation strategy, especially relevant in the case of S. cerevisiae and L. thermotolerans mixed fermentation, that has an outstanding release of 4MSP thiol. Although our results stem from a specific experimental approach that differs from real winemaking situations, these findings reveal the potential of unravelling the specific role of different yeast species, thiol precursors and their interaction, to improve wine production processes in the context of wine aroma enhancement.

A Droplet-Based Microfluidic Platform for High-Throughput Culturing of Yeast Cells in Various Conditions.

Yeast plays a significant role in a variety of fields. In particular, it is extensively used as a model organism in genetics and cellular biology studies, and is employed in the production of vaccines, pharmaceuticals, and biofuels. Traditional "bulk"-based studies on yeast growth often overlook cellular variability, emphasizing the need for single-cell analysis. Micro-droplets, tiny liquid droplets with high surface-area-to-volume ratios, offer a promising platform for investigating single or a small number of cells, allowing precise control and monitoring of individual cell behaviors. Microfluidic devices, which facilitate the generation of micro-droplets, are advantageous due to their reduced volume requirements and ability to mimic in vivo micro-environments. This study introduces a custom-designed microfluidic device to encapsulate yeasts in micro-droplets under various conditions in a parallel manner. The results reveal that optimal glucose concentrations promoted yeast growth while cycloheximide and Cu2+ ions inhibited it. This platform enhances yeast cultivation strategies and holds potential for high-throughput single-cell investigations in more complex organisms.

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.

Antimicrobial Activity of Leaf Aqueous Extract of Schinus polygamus (Cav.) Cabrera against Pathogenic Bacteria and Spoilage Yeasts.

The antimicrobial activity of an aqueous extract of the leaves of Schinus polygamus (cav.) Cabrera against microorganisms of food importance was evaluated. First, the leaf aqueous extract of Schinus polygamus was characterized, quantifying hydroxycinnamic acids and phenolic compounds. Then, a battery of strains was tested, including Escherichia coli ATCC 25922, Salmonella Typhimurium ATCC 14028, and Listeria monocytogenes ATCC 13932. Also, we tested wine spoilage yeasts such as Brettanomyces bruxellensis LAMAP2480, B. bruxellensis LAMAP1359, B. bruxellensis CECT1451, and Pichia guilliermondii NPCC1051. Tests were conducted using the kinetic curve of growth and cell viability counts. The results indicate that with 10% v/v of concentrated extract, it is possible to observe growth inhibition of all microorganisms studied, with statistically significant differences during the whole measurement time (70 h for bacteria and 145 h for yeast).

Effects of Fermentation with Kombucha Symbiotic Culture of Bacteria and Yeasts on Antioxidant Activities, Bioactive Compounds and Sensory Indicators of Rhodiola rosea and Salvia miltiorrhiza Beverages.

Kombucha is a well-known fermented beverage traditionally made from black tea infusion. Recent studies have focused on finding alternative materials to create novel kombucha beverages with various health benefits. In this study, we prepared and evaluated two novel kombucha beverages using Rhodiola rosea and Salvia miltiorrhiza as materials. The effects of fermentation with the residue of these plants on the kombucha were also investigated. The antioxidant activities, total phenolic contents, and concentrations of the bioactive compounds of the kombucha beverages were determined by the Trolox equivalent antioxidant capacity test, ferric-reducing antioxidant power test, Folin-Ciocalteu method, and high-performance liquid chromatography, respectively. The results revealed that the kombucha beverages made with Rhodiola rosea and Salvia miltiorrhiza had strong antioxidant capacities and abundant phenolic contents. Additionally, the kombucha fermented with Rhodiola rosea residue had higher FRAP, TEAC and TPC values than that fermented without residue. On the other hand, the Salvia miltiorrhiza kombucha fermented with residue had similar FRAP and TEAC values but lower TPC values compared to that fermented without residue. The correlation analysis showed that gallic acid, salidroside, and tyrosol were responsible for the antioxidant abilities and total phenolic contents of the Rhodiola rosea kombucha, and salvianolic acid A and salvianolic acid B contributed to the antioxidant abilities of the Salvia miltiorrhiza kombucha. Furthermore, the kombucha fermented with Rhodiola rosea residue had the highest sensory scores among the kombucha beverages studied. These findings suggest that Rhodiola rosea and Salvia miltiorrhiza are suitable for making novel kombucha beverages with strong antioxidant abilities and abundant phenolic contents, which can be used in preventing and managing oxidative stress-related diseases.

Bioprocess Optimization by Taguchi Design and Response Surface Analysis for Obtaining Active Yeast Used in Vinification

This study presents the behavior of the identified yeast strain S. cerevisiae, isolated from Busuioacă de Bohotin grapes from the Pietroasa winery, during the process of fermentation to obtain dry active yeast biomass for the winemaking process. In this respect, we promoted an optimization strategy for obtaining active dry yeast biomass. The cultivation conditions for micropilot fermentation (temperature, pH, carbon source, and nitrogen source) were selected and designed according to a Taguchi design with four factors and three levels. Reproducibility testing was conducted under specific fermentation parameters: temperature (32 °C), pH (4.5), carbon source (12%), and nitrogen source (0.7%). Following the optimization process, two combinations of cultivation parameters were selected, and one of them, based on the results, was selected for further analysis. Quantitative data were obtained, showing dry yeast biomass (DCW) at 1.39 g/100 mL and protein content at 45.57%. The active yeast was then used in the winemaking process for Tămâioasă Românească and Busuioacă de Bohotin varieties at Pietroasa winery for validation. This optimization aims to facilitate easy and rapid production of fresh wine yeast tailored to the local winemaking practices of Pietroasa winery, with real application potential in other viticultural areas, aligning with the terroir concept.

Evaluation of the Nonlinear Response of KCl Solution to High-Amplitude Stimulation Using a Four-Point Cylindrical Gold Electrode.

A four-electrode system and high-voltage amplitude stimulation (with a voltage of more than 1 V) are always used for nonlinear spectroscopy of biological solutions like yeast suspension. Multiple studies indicate that yeast membrane activities are the primary source of the harmonics detected from yeast suspensions. However, other investigations have demonstrated that at least part of the harmonics are mainly due to the nonlinear impedance of the electrode-electrolyte interface. Here, we want to find a mathematical model to analyze the odd harmonics generated by the interface. The KCl solution is the basis of the essential medium used in yeast culture. No study models the nonlinear behavior of the KCl solution using four-electrode. We used a sinusoidal voltage with a low frequency and a high amplitude to test an ionic solution with 1, 30, and 130 mM of potassium chloride. The voltage had no direct current component. Here, we show that the relationship between the amplitude of the excitation voltage and the third harmonic of the current can be accurately represented using the Butler-Volmer equation. Additionally, we analyze the impact of potassium chloride (KCl) concentration on the behavior of the third harmonic. This paper presents a method and model that can be used for nonlinear spectroscopy studies of biological solutions using a four-electrode system with the aim of deriving information from cells without considering the interface effect.

Effects of yeast culture supplementation on milk yield, rumen fermentation, metabolism, and bacterial composition in dairy goats.

The effects of yeast culture (YC) on dairy goat milk yield and potential effects of rumen microbial population changes on rumen fermentation are poorly understood. This study aimed to evaluate the effects of YC on milk yield and rumen fermentation in dairy goats and explore the potential microbial mechanisms. Forty Laoshan dairy goats with a weight of 51.23 ± 2.23 kg and daily milk yield of 1.41 ± 0.26 kg were randomly divided into 4 groups: control (no YC), YC1 (10 g/day per goat), YC2 (25 g/day per goat), and YC3 (40 g/day per goat). The pre-feeding period was 15 days, and the official period was 60 days. Laoshan dairy goats were milked twice daily, and the individual milk yield was recorded. On the last day of the official period, rumen fluid was collected to measure rumen fermentation, perform quantitative polymerase chain reaction (PCR), and detect metabolites. Compared to the control group, the YC group had greater milk yield; higher acetic acid, butyric acid, and total volatile fatty acid contents; and lower ammonia-N (NH3-N) content in the rumen (p < 0.05). YC increased the abundance of Clostridia_UCG-014 and Paraprevotella (p < 0.05). Differential metabolites L-leucine and aspartic acid were screened. This study revealed the microbial mechanisms linking the relative abundance of Paraprevotella and Clostridia_UCG-014 to L-leucine and aspartic acid utilization. These results describe the potential benefits of supplementing 10 g/day per goat YC in the diets of Laoshan dairy goats for improving the rumen environment and milk yield.

Quality improvement of nata bioedible film using Acetobacter xylinum and kombucha symbiotic culture of bacteria and yeast as starter

The trend towards bio-edible films is being promoted to reduce the use of plastic packaging, specifically as primary packaging for food. Therefore, this study aimed to improve the quality of nata as bioedible film by using a starter during fermentation to produce microbial metabolites such as the cellulose matrix, “nata”. The fermentation of nata was carried out using coconut water as substrate, while the starter used single culture (Acetobacter xylinum) and multiculture (kombucha symbiotic culture of bacteria and yeast (SCOBY)). The starter (15% v/v) was inoculated on the coconut water substrate which had been formulated and adjusted at pH 3-4, then incubated for 14, 18 and 22 days. Fermentation using the kombucha SCOBY for 22 days of fermentation resulted in good quality nata biofilm. The results showed that the yield of nata reached 35.44% w/v with 14.76 mm of thickness, 5 cm of diameter, 0.86 N/nm2 of elasticity, 34.02-52.01 MPa of tensile strength, and 2.3-2.6% of elongation. The functional groups profile of nata edible film was OH, CH, C=C, and aromatic rings. The results of this study showed that kombucha SCOBY has good potential as a starter for producing bioedible films

Multidisciplinary advances in kombucha fermentation, health efficacy, and market evolution.

Kombucha, a fermented tea beverage, has seen a significant rise in global popularity. This increase is attributed to its reported health benefits and extensive cultural heritage. The comprehensive review examines kombucha through microbiology, biochemistry, and health sciences, highlighting its therapeutic potential and commercial viability. Central to kombucha production is the symbiotic culture of bacteria and yeasts (SCOBY), which regulates a complex fermentation process, resulting in a bioactive-rich elixir. The study examines the microbial dynamics of SCOBY, emphasizing the roles of various microorganisms. It focuses the contributions of acetic acid bacteria, lactic acid bacteria, and osmophilic yeasts, including genera such as Saccharomyces, Schizosaccharomyces, Zygosaccharomyces, Brettanomyces/Dekkera, and Pichia. These microorganisms play crucial roles in producing bioactive compounds, including organic acids, polyphenols, and vitamins. These bioactive compounds confer therapeutic properties to kombucha. These properties include antioxidant, antimicrobial, anti-inflammatory, antidiabetic, antihypertensive, cancer prevention, hepatoprotective, and detoxifying effects. The review also explores the growing market for kombucha, driven by consumer demand for functional beverages and opportunities for innovative product development. It emphasizes the necessity of standardized production to ensure safety and validate health claims. Identifying research gaps, the review highlights the importance of clinical trials to verify therapeutic benefits. Ultimately, this study integrates traditional knowledge with scientific research, providing directions for future studies and commercial expansion, emphasizing the role of kombucha in health and wellness.

Identification of ERAD-dependent degrons for the endoplasmic reticulum lumen.

Degrons are minimal protein features that are sufficient to target proteins for degradation. In most cases, degrons allow recognition by components of the cytosolic ubiquitin proteasome system. Currently, all of the identified degrons only function within the cytosol. Using Saccharomyces cerevisiae, we identified the first short linear sequences that function as degrons from the endoplasmic reticulum (ER) lumen. We show that when these degrons are transferred to proteins, they facilitate proteasomal degradation through the ERAD system. These degrons enable degradation of both luminal and integral membrane ER proteins, expanding the types of proteins that can be targeted for degradation in budding yeast and mammalian tissue culture. This discovery provides a framework to target proteins for degradation from the previously unreachable ER lumen and builds toward therapeutic approaches that exploit the highly-conserved ERAD system.

Precise evaluation of the nutritional value of yeast culture and its effect on pigs fed low-protein diets

The purpose of the present study was to assess the nutritional value of yeast culture (YC) and to explore the effect of YC on growth performance and health of piglets fed low-protein (LP) diets. In Exp.1, 12 growing barrows were allocated into control diet and YC diet treatments to determine the available energy of YC. Results showed that the digestible energy and metabolizable energy of YC are 12.12 and 11.66 MJ/kg dry matter (DM), respectively. In Exp.2, 12 growing barrows were surgically equipped with a T-cannula near the distal ileum and were assigned to 2 dietary treatments (nitrogen-free diet and YC diet), and the amino acid digestibility of YC was determined. In Exp.3, a total of 96 weaned piglets were randomly divided into 4 treatments, including LP basal diet (Basal), Basal+0.5% YC (0.5%YC), Basal+1.0% YC (1%YC), and Basal+1.5% YC (1.5%YC). The results were as follows: YC supplementation linearly improved the weight gain and feed intake ratio (P < 0.001), linearly increased the activity of glutathione peroxidase on d 14 (P = 0.032) and linearly decreased the concentration of malondialdehyde on d 14 (P = 0.008) and d 32 (P = 0.004) in serum, and linearly decreased the concentration of total short-chain fatty acid on d 14 in feces (P = 0.045). Compared with other treatments, 1.5%YC group showed a greater abundance of various probiotics, such as Prevotellaceae, Prevotella and Turicibacter. In Exp.4, twelve growing barrows with an ileal T-cannula were randomly assigned to Control and 1.5%YC treatments to clarify the impact of YC supplementation on nitrogen balance and nutrient digestibility. Results showed that YC had no significant effect on nitrogen efficiency and nutrient digestibility, except for trend of reducing the total tract digestibility of organic matter (P = 0.067). In conclusion, the present study assessed the digestible and metabolizable energy values (12.12 and 11.66 MJ/kg DM, respectively) and standardized ileal digestibility of amino acid (from 43.93% to 82.65%) of YC in pig feed and demonstrated that moderate supplementation of YC (1.5% of diet) can effectively improve feed conversion efficiency, enhance antioxidant capacity, and promote a balanced gut microbiota in piglets.

Study on the changes during the fermentation of the wine prepared from palm (Phoenix sylvestris) sap

The sugary sap of different palm trees is fermented to create palm wine, an alcoholic beverage. This work was aimed at studying the changes that occur during the fermentation process of wine made from the sap of the wild date palm species Phoenix sylvestris. At first, the best age of the palm tree was determined by observing total soluble solid and sap yield for 24 h and was found to be middle-aged palm plants (15–40 years old). Pure wine yeast (Saccharomyces cerevisiae SC22) and a natural starter culture were added to the palm saps, adjusting the total soluble solid (TSS) to 21.5° brix (°Bx). Total titratable acidity, pH, volatile acidity, reducing sugar, non-reducing sugar, total sugar, alcohol content, ester content, and aldehyde contents were the parameters under investigation. The statistical analysis showed significant (p ≤ 0.05) changes in the physico-chemical and volatile constituents of palm sap during the fermentation process in both systems. Sensory evaluation revealed that palm wine fermented with pure yeast culture was significantly superior to natural, spontaneously fermented wine. The acceptability test showed that the ideal characteristics of palm wine are cloudy in appearance, fruity in aroma, and sweet in taste.

Cloning and Optimization of Intracellular Expression of Human Interferon β-1a in Pichia pastoris GS115.

Interferon-beta (IFN-β) is a cytokine with a wide range of biological and pharmaceutical applications, including multiple sclerosis (MS), cancer, some autoimmune disorders, and viral infectious diseases. Thus, many studies have been performed to develop novel strategies for the high-yield production of functional IFN-β in a cost-effective approach. Here, we aimed to improve the intracellular expression of IFN-β-1a in Pichia pastoris. The gene of IFN-β-1a was successfully sub-cloned into the pPICZA vector. The recombinant vector was transfected to P. pastoris GS115 cells by electroporation. After screening positive P. pastoris transformants, the expression of IFN-β-1a was evaluated and the cultivation conditions, including temperature, time of incubation, and methanol concentration, were optimized. The protein expression levels were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The double digestion with EcoRI and XhoI restriction enzymes and sequence analysis confirmed the correct sub-cloning of the IFN-β-1a gene into pPICZA. SDS-PAGE analysis showed that the highest level of IFN-β-1a (25 mg per 1 L of yeast culture) was produced with 2% methanol at 28°C after 72 h incubation. Optimization of cultivation conditions for intracellular expression of IFN-β-1a was successfully performed. This approach can be generally applied to improve the production yield and quality of other recombinant proteins in P. pastoris.

The Effect of Fermentation Time Variation on Antioxidant Activity of Fermented Robusta Coffee Leaves (Coffea canephora Pierre ex A.Froehner) Brew

Antioxidants are electron-donor compounds that can delay, slow down and prevent free radical reactions. Fermentation is a chemical change that occurs due to the activity of microorganisms which can increase the amount of phenols in tea thereby increasing its antioxidant activity. Fermentation is carried out with the help of a Symbiotic Culture of Bacteria and Yeast (SCOBY) culture, which is commonly used in the production of kombucha. Kombucha is a fermented tea drink produced from leaves that contain high phenols such as robusta coffee leaves (Coffea canephora Pierre ex A.Froehner). Factors that affect the fermentation process are pH and fermentation time. This study aimed to explain the effect of fermentation time on the antioxidant activity of fermented robusta coffee leaves brew. Brewed robusta coffee leaves were made by brewing robusta coffee leaves simplicia powder, adding kombucha liquid starter, then fermented for 0; 3; 7; 14; and 21 days, then pH and antioxidant activity was measured using UV-Visible spectrophotometer using the 2,2-dipheny-1,1-picrylhydrazyl (DPPH) at a wavelength of 516 nm. This study showed that 0; 3; 7; 14; and 21 days fermented robusta coffee leaves brew have pH of 6,07; 4,62; 4,07; 3,21; and 2,74, and IC50 values is 285,1645 μg/ml; 219,7012 μg/ml; 192,2325 μg/ml; 187,0434 μg/ml; and 173,9249 µg/ml. There is an effect of fermentation time variation on the antioxidant activity of fermented robusta coffee leaves brew. Keyword: Antioxidant, Fermentation, pH, Robusta coffee leaves, Time variation

Antimicrobial activities of silver nanoparticles synthesized from Helichrysum italicum (Roth) G. Don

In our study, the ability to produce silver nanoparticles (AgNP) by green synthesis method using Helichrysum italicum (Roth) G. Don. plant was investigated. The color of the extract changed from yellow to brown, proving the presence of AgNPs. Transmission electron microscope (TEM), scanning electron microscopy (SEM), and Energy-dispersive spectra (EDS), FT-IR and Uv-Vis analysis were used to characterized AgNPs. AgNPs ranged from 20 nm to 40 nm in size and were mostly spherical and cubic structures. The antimicrobial potential of AgNPs was tested against seven bacterial and one yeast cultures by disc diffusion and minimum inhibitory concentration (MIC) methods. AgNPs significantly inhibited all test cultures. The highest antimicrobial activities of plant extract and AgNP were found against Proteus vulgaris ATCC 13315.