Yeast Culture Research Articles

Physical Modifications of Kombucha‐Derived Bacterial Nanocellulose: Toward a Functional Bionanocomposite Platform

AbstractSustainable functionalization of bacterial cellulose for cost‐effective bionanocomposites with desired properties has received growing attention in recent years. This article presents the results of work aimed at obtaining bionanocomposite materials based on bacterial cellulose, a natural and eco‐friendly material. Bacterial cellulose obtained from the Kombucha symbiotic culture of bacteria and yeast (SCOBY) fermentation process is functionalized by embedding with diatom frustules, silver nanoparticles (AgNPs), and poly(3,4‐ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS). The effects of functionalization on mechanical, optical, plasmonic, electrical, chemiluminescent, and antimicrobial properties are evaluated. Morphological characteristics of the nanocomposites are studied using electron microscopy. Addition of diatom frustules introduced into the SCOBY culture media results in bionanocomposite materials with enhanced tensile strength and increased ultraviolet (UV) blockage properties. In situ functionalization of bacterial cellulose with AgNPs tunes plasmonic and chemiluminescent properties, revealing the biosensing potential of the material. Modified bacterial cellulose shows antimicrobial activity in experiments with gram‐positive and gram‐negative bacteria. Dual functionalization of bacterial cellulose with PEDOT:PSS and AgNPs results in improved electrical conductivity of the bionanocomposite. Overall, bottom‐up physical functionalization approaches and the resulting bionanocomposite materials will open up new opportunities for the low‐cost production of green materials and contribute to the development of a sustainable economy.

Cultivation of yeasts on liquid digestate to remove organic pollutants and nutrients and for potential application as co-culture with microalgae

In this study, the growth of yeast and yeast-like fungi in the liquid digestate from vegetable wastes was investigated in order to remove nutrients and organic pollutants, and for their application as co-culture members with green microalgae. The studied yeast strains were characterized for their assimilative and enzymatic profiles as well as temperature requirements. In the first experimental stage, the growth dynamics of each strain were determined, allowing to select the best yeasts for further studies. In the subsequent stage, the ability of selectants to remove organic pollutants was assessed. Different cultivation media containing respectively 1:3, 1:1, 3:1 vol ratio of liquid digestate and the basal minimal medium were used. Among all tested yeast strains, Rhodotorula mucilaginosa DSM 70825 showed the most promising results, demonstrating the highest potential for removing organic substrates and nutrients. Depending on the medium, this strain achieved 50–80% sCOD, 45–60% tVFAs, 21–45% TN, 33–52% PO43− reduction rates. Similar results were obtained for the strain Candida sp. OR687571. The high nutrient and organics removal efficiency by these yeasts could likely be linked to their ability to assimilate xylose (being the main source of carbon in the liquid digestate). In culture media containing liquid digestate, both yeast strains achieved good viability and proliferation potential. In the liquid digestate medium, R. mucilaginosa and Candida sp. showed vitality at the level of 51.5% and 45.0%, respectively. These strains seem to be a good starting material for developing effective digestate treatment strategies involving monocultures and/or consortia with other yeasts or green microalgae.

Enhancement of biochemical parameters and enzyme activity in solid-state fermented and biofortified maize cobs utilizing yeasts and plant extracts

This study investigated the impact of solid-state fermentation and biofortification of maize cobs using yeast and plant extracts on their biochemical properties and enzyme activity. Maize cobs were fermented with extracts from elephant grass, cassava leaves, or Siam weed, combined with baker's yeast or palm wine yeast. In vitro assays revealed increased l-lysine concentration (up to 8.6 mg/g), soluble protein content (up to 27.4 mg/g), total phenolic content (up to 15.5 μg GAE/g), total flavonoid content (up to 23.4 μg CE/g), and xylanase activity (up to 25.7 Units) compared to controls. Glucose concentration decreased (to 0.8 mg/mL), indicating efficient utilization. Amylase and protease activities varied, with some combinations showing higher enzymatic activity (amylase at 35.4 Units, protease at 39.2 Units). These results underscore the potential of solid-state fermentation and biofortification to enhance the nutritional quality of agro-residues, offering novel insights into sustainable food production and waste utilization strategies.

Enhancing coffee sensory quality: unleashing synergistic effects of selected microorganism starter cultures and ohmic heating technology

Abstract The sensory characteristics of coffee play a pivotal role in determining its consumer appeal and global market potential. The flavor quality of coffee is influenced by a range of factors, including genetic attributes, geographic variables, agricultural practices, and post-harvest processes. Among these, the post-harvest phase, which includes fermentation, stands out as a critical determinant of coffee quality. Notably, controlled wet fermentation techniques and the introduction of specialized starter cultures have demonstrated a significant capacity to enhance the flavor and unique sensory profiles of coffee, capturing the interest of consumers. Utilizing yeast cultures, such as Saccharomyces cerevisiae and Torulaspora delbrueckii, alongside lactic acid bacteria like Lactobacillus sp., has been scientifically substantiated as a means to elevate coffee quality. The application of ohmic heating technology during the coffee fermentation process presents an innovative approach to maintaining the fermentation process at the desired temperature. When synergistically applied in tandem with carefully selected microorganism starter cultures, this approach holds the promise of delivering a substantial enhancement in coffee quality.

Potential symbiotic culture of bacteria and yeast (SCOBY) jackfruit leaves as an active ingredient for skin pathogens infection treatment

Potential symbiotic culture of bacteria and yeast (SCOBY) jackfruit leaves as an active ingredient for skin pathogens infection treatment

Influence and metabolomic basis of an indigenous yeast CECA, from Ningxia wine region of China, on the aroma and flavor of Cabernet Sauvignon wines

In this study, three fermentation treatments of spontaneous fermentation (SF), direct inoculation of CECA (YF), and inoculation with CECA after addition of dimethyl dicarbonate (YDF) were carried out. Multivariate statistical analysis approved that CECA inoculation significantly influenced the composition of 141 metabolites (15 volatile organic compounds (VOCs) and 126 non-VOCs), mainly consisting of 36 acids and derivatives and 25 lipids and lipid-like molecules. YF and YDF wines exhibited similar correlations with aroma types, while there were differences in the kinds and number of VOCs. Moreover, CECA-inoculated fermentation was more favorable to the formation of aftertaste-A, umami, sourness, and richness. The KEGG metabolic pathway analysis indicated that the inoculation strategy significantly affected the amino acid metabolism. The antimicrobial treatment effectively enhanced bitterness, astringency, umami and saltiness while reducing acidity. Further studies are needed to assess the effects of antimicrobial treatment on lipid metabolism.

Integrated omics of Saccharomyces cerevisiae CENPK2-1C reveals pleiotropic drug resistance and lipidomic adaptations to cannabidiol

Yeast metabolism can be engineered to produce xenobiotic compounds, such as cannabinoids, the principal isoprenoids of the plant Cannabis sativa, through heterologous metabolic pathways. However, yeast cell factories continue to have low cannabinoid production. This study employed an integrated omics approach to investigate the physiological effects of cannabidiol on S. cerevisiae CENPK2-1C yeast cultures. We treated the experimental group with 0.5 mM CBD and monitored CENPK2-1C cultures. We observed a latent-stationary phase post-diauxic shift in the experimental group and harvested samples in the inflection point of this growth phase for transcriptomic and metabolomic analysis. We compared the transcriptomes of the CBD-treated yeast and the positive control, identifying eight significantly overexpressed genes with a log fold change of at least 1.5 and a significant adjusted p-value. Three notable genes were PDR5 (an ABC-steroid and cation transporter), CIS1, and YGR035C. These genes are all regulated by pleiotropic drug resistance linked promoters. Knockout and rescue of PDR5 showed that it is a causal factor in the post-diauxic shift phenotype. Metabolomic analysis revealed 48 significant spectra associated with CBD-fed cell pellets, 20 of which were identifiable as non-CBD compounds, including fatty acids, glycerophospholipids, and phosphate-salvage indicators. Our results suggest that mitochondrial regulation and lipidomic remodeling play a role in yeast’s response to CBD, which are employed in tandem with pleiotropic drug resistance (PDR). We conclude that bioengineers should account for off-target product C-flux, energy use from ABC-transport, and post-stationary phase cell growth when developing cannabinoid-biosynthetic yeast strains.

Enzymatic Activity of Fungi for Hydrolysis of Wheat Bran and Cultivation of Oleaginous Yeasts

Sustainable alternatives to petrochemical raw materials in oleochemical, laundry detergent and personal care industry need to be implemented. Presently, palm oil is the predominant cost-efficient option. While it is a renewable source, it has negative environmental and socioeconomic implications. Therefore, the request for ecologically certified and palm oil-free cleaning agents and household care products is rapidly growing, supported by concepts of waste-free biorefinery. Oleaginous yeasts are currently the most promising source for microbial lipid production. Additionally, wheat bran is a cereal milling by-product, which is available in high quantities and provides an excellent feedstock. Enzymatically generated wheat bran hydrolysate provides an excellent microbial cultivation medium for the sustainable production of high value biological additives. Indeed, wheat bran consists predominantly of lignocellulose, and therefore of cellulose, hemicellulose and lignin, which can be hydrolyzed by established enzyme systems. Currently, such enzymatic systems are mainly obtained for the market from genetically modified organisms (GMO). This represents a problem because GMO products or process components cannot be included in eco-certified household care formulations. Therefore, innovative GMO-free biotechnological processes need to be implemented, as well as the isolation of novel GMO-free hydrolase enzyme systems. Fungal strains are an interesting source for the efficient screening and isolation GMO-free hydrolases (see figure).This contribution is part of a project, which aims to use a holistic biotechnological approach to convert mill residues into customized specialty ingredients of eco-certified detergents. The project focuses on three main tasks:investigation of a microbial group of eight Ascomycete fungi,production of enzymes for hydrolysis of wheat bran, anduse of the resulting biomass hydrolysate to cultivate oleaginous yeasts.Hydrolase producing fungal strains were cultivated on different substrates to investigate their ability to generate specific enzyme systems capable of hydrolyzing wheat bran. The activity of the enzymes was qualitatively and quantitatively characterized to determine the optimal strains for the degradation of wheat bran and suited to implement a large-scale process. Supernatants of different fungal strains cultivations were tested in the hydrolysis step to compare the hydrolysis performances of differently active enzyme mixes. The biomass hydrolysate was used to cultivate lipid generating yeasts to investigate their ability to grow on the microbial cultivation medium.

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

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

Enhancing Selenium Accumulation in Rhodotorula mucilaginosa Strain 6S Using a Proteomic Approach for Aquafeed Development.

It is known that selenium (Se) is an essential trace element, important for the growth and other biological functions of fish. One of its most important functions is to contribute to the preservation of certain biological components, such as DNA, proteins, and lipids, providing protection against free radicals resulting from normal metabolism. The objective of this study was to evaluate and optimize selenium accumulation in the native yeast Rhodotorula mucilaginosa 6S. Sodium selenite was evaluated at different concentrations (5-10-15-20-30-40 mg/L). Similarly, the effects of different concentrations of nitrogen sources and pH on cell growth and selenium accumulation in the yeast were analyzed. Subsequently, the best cultivation conditions were scaled up to a 2 L reactor with constant aeration, and the proteome of the yeast cultured with and without sodium selenite was evaluated. The optimal conditions for biomass generation and selenium accumulation were found with ammonium chloride and pH 5.5. Incorporating sodium selenite (30 mg/L) during the exponential phase in the bioreactor after 72 h of cultivation resulted in 10 g/L of biomass, with 0.25 mg total Se/g biomass, composed of 25% proteins, 15% lipids, and 0.850 mg total carotenoids/g biomass. The analysis of the proteomes associated with yeast cultivation with and without selenium revealed a total of 1871 proteins. The results obtained showed that the dynamic changes in the proteome, in response to selenium in the experimental medium, are directly related to catalytic activity and oxidoreductase activity in the yeast. R. mucilaginosa 6S could be an alternative for the generation of selenium-rich biomass with a composition of other nutritional compounds also of interest in aquaculture, such as proteins, lipids, and pigments.

Beneficial effects of plant growth-promoting yeasts (PGPYs) on the early stage of growth of zucchini plants

The use of microbes capable of beneficially interacting with plants is essential for advancing climate-smart agriculture. This approach aims to reduce chemical use while simultaneously enhancing crop productivity. This implies efforts to optimize the criteria for selecting potential plant growth promoters (PGPs), focusing also on yeasts, only recently investigated for their PGP potential. The present study employed a set of Ascomycetes and Basidiomycetes yeasts to test their PGP properties on zucchini (Cucurbita pepo L.), chosen as a fast-growing plant with a vast economical interest. Yeasts were tested alone and as consortium. Seed inoculation with yeasts boosted the early phase of growth of the zucchini plants, primarily affecting the root development. Three strains belonging to the species Schwanniomyces etchellsii, Zygotorulaspora florentina and Holtermanniella festucosa induced a strong and significant enhancement of weight and length of both epi- and hypogeal parts of the plant. Furthermore, the presence of yeasts induced strain-specific modulations in the biochemical profiles of soil, primarily detected in the rhizosphere. This suggests an active interaction between the roots and the inoculated yeast cultures.

Studying the efficiency of Gaprin application in sterlet farming

In the course of the experiment, the influence of new feed additives created on the basis of Gaprin and representing biomass of inactivated cell culture of non-pathogenic methane-oxidizing yeasts or bacteria on sturgeon fish productivity indicators was studied. The research was carried out in the Krasnodar Territory on sterlet fingerlings. The experiment lasted for 60 days. According to the scheme of the experiment, the 1st (control) group of fish received complete feeds without additives, i.e. containing 100.0% fish meal. In group 2 feeding, 50.0% fishmeal was replaced with 50.0% Gaprin. Group 3 diet contained 25.0% fishmeal and 75.0% Gaprin. In the feed for group 4, fish meal and Gaprin were in the ratio of 75/25%. In Group 5, fish meal was completely replaced by Gaprin, in this case the amount of additive was 10.5 kg. Sterlet fingerlings from Group 6 received feed with 10.0% Gaprin. The initial live weight in all groups was similar at 30 g. However, the inclusion of Gaprin in the diet by reducing the amount of fish meal contributed to an increase in the final live weight of sterlets in the experimental groups by 4.4–19.4% compared to the control. The exceptions were groups 5 and 6, which received 100.0 and 90.0% Gaprin instead of fishmeal. Final live weight in these groups was lower by 2.3 and 0.9%, respectively. Feed costs per 1 kg of live weight gain in the groups with partial or complete replacement of fish meal were lower relative to the 1st group (from 7.0 to 17.9%,).Inclusion of Gaprin in complete feeds contributed to the increase in profit from 553.00 to 1785.00 rubles.

Comparison of Anti-Inflammatory and Antibacterial Properties of Raphanus sativus L. Leaf and Root Kombucha-Fermented Extracts.

In the cosmetics industry, the extract from Raphanus sativus L. is fermented using specific starter cultures. These cosmetic ingredients act as preservatives and skin conditioners. Kombucha is traditionally made by fermenting sweetened tea using symbiotic cultures of bacteria and yeast and is used in cosmetic products. The aim of this study was to evaluate the cosmetic properties of radish leaf and root extract fermented with the SCOBY. Both unfermented water extracts and extracts after 7, 14, and 21 days of fermentation were evaluated. The analysis of secondary plant metabolites by UPLC-MS showed higher values for ferments than for extracts. A similar relationship was noted when examining the antioxidant properties using DPPH and ABTS radicals and the protective effect against H2O2-induced oxidative stress in fibroblasts and keratinocytes using the fluorogenic dye H2DCFDA. The results also showed no cytotoxicity to skin cells using Alamar Blue and Neutral Red tests. The ability of the samples to inhibit IL-1β and COX-2 activity in LPS-treated fibroblasts was also demonstrated using ELISA assays. The influence of extracts and ferments on bacterial strains involved in inflammatory processes of skin diseases was also assessed. Additionally, application tests were carried out, which showed a positive effect of extracts and ferments on TEWL and skin hydration using a TEWAmeter and corneometer probe. The results obtained depended on the concentration used and the fermentation time.

Improvement of the recombinant phytase expression by intermittent feeding of glucose during the induction phase of methylotrophic yeast Pichia pastoris.

For growth of methylotrophic yeast, glycerol is usually used as a carbon source. Glucose is used in some cases, but not widely consumed due to strong repressive effect on AOX1 promoter. However, glucose is still considered as a carbon source of choice since it has low production cost and guarantees growth rate comparable to glycerol. In flask cultivation of the recombinant yeast, Pichia pastoris GS115(pPIC9K-appA38M), while methanol induction point(OD600) and methanol concentration significantly affected the phytase expression, glucose addition in induction phase could enhance phytase expression. The optimal flask cultivation conditions illustrated by Response Surface Methodology were 10.37 OD600 induction point, 2.02h before methanol feeding, 1.16% methanol concentration and 40.36μL glucose feeding amount(for 20mL culture volume) in which the expressed phytase activity was 613.4 ± 10.2U/mL, the highest activity in flask cultivation. In bioreactor fermentation, the intermittent glucose feeding showed several advantageous results such as 68h longer activity increment, 149.2% higher cell density and 200.1% higher activity compared to the sole methanol feeding method. These results implied that remaining glucose at induction point might exhibit a positive effect on the phytase expression. Glucose intermittent feeding could be exploited for economic phytase production and the other recombinant protein expression by P. pastoris GS115.

Gradient boosted regression as a tool to reveal key drivers of temporal dynamics in a synthetic yeast community.

Microbial communities are vital to our lives, yet their ecological functioning and dynamics remain poorly understood. This understanding is crucial for assessing threats to these systems and leveraging their biotechnological applications. Given that temporal dynamics are linked to community functioning, this study investigated the drivers of community succession in the wine yeast community. We experimentally generated population dynamics data and used it to create an interpretable model with a gradient-boosted regression tree approach. The model was trained on temporal data of viable species populations in various combinations, including pairs, triplets, and quadruplets, and was evaluated for predictive accuracy and input feature importance. Key findings revealed that the inoculation dosage of non-Saccharomyces species significantly influences their performance in mixed cultures, while Saccharomyces cerevisiae consistently dominates regardless of initial abundance. Additionally, we observed multispecies interactions where the dynamics of Wickerhamomyces anomalus were influenced by Torulaspora delbrueckii in pairwise cultures, but this interaction was altered by the inclusion of S. cerevisiae. This study provides insights into yeast community succession and offers valuable machine learning-based analysis techniques applicable to other microbial communities, opening new avenues for harnessing microbial communities.

Impact of different fermentation times on the microbiological, chemical, and sensorial profile of coffees processed by self-induced anaerobiosis fermentation.

Variation in fermentation time may be an essential alternative to provide coffee beverages with different and unique sensory profiles. This work investigated the microbiological, chemical, and sensory changes in coffees submitted to different fermentation durations (0, 24, 48, 72, and 96h). Self-induced anaerobiosis fermentation (SIAF) was used, and two treatments were performed: spontaneous fermentation and inoculation with S. cerevisiae CCMA0543. Microbiological analyses were performed, and the permanence of the inoculum was monitored. Chromatography (sugars, organic acids, and volatile compounds) was analyzed, and sensory analysis (temporal dominance of sensations - TDS) was performed. A total of 228 isolates were identified during spontaneous fermentation. The dominant bacteria and yeasts were Leuconostoc mesenteroides, Lactiplantibacillus plantarum, Staphylococcus warneri, Bacillus sp., Torulaspora delbrueckii, Hanseniaspora uvarum, and Meyerozyma caribbica. High concentrations of citric (18.67 mg.g- 1) and succinic (5.04 mg.g- 1) acids were detected at 96h in SIAF fermentation. One hundred twenty-one volatile compounds were detected, but 22 were detected only in inoculated coffees. In spontaneous fermentation, 48h of fermentation showed woody notes, while 72h showed chestnuts. However, in the inoculated coffee, 72h of fermentation showed high fruity dominance, and 96h of fermentation was the only one with herbaceous notes. In addition, yeast inoculation increased the intensity of caramel notes in the first 48h and increased the fruity flavor after 72h of fermentation. Therefore, the type of fermentation (with or without inoculation) and the chosen fermentation time will depend on the sensorial profile the producer intends to obtain.

Dietary 5-hydroxytryptophan improves sheep growth performance by enhancing ruminal functions, antioxidant capacity, and tryptophan metabolism: in vitro and in vivo studies.

Hydroxytryptophan (5-HTP) can regulate the synthesis of 5-Hydroxytryptamine (5-HT) and melatonin (MT). In a previous metabolome analysis, we found that 5-HTP is an effective ingredient in yeast culture for regulating rumen fermentation. However, research on the effect of this microbial product (5-HTP) as a functional feed additive in sheep production is still not well explained. Therefore, this study examined the effects of 5-HTP on sheep rumen function and growth performance using in vitro and in vivo models. A two-factor in vitro experiment involving different 5-HTP doses and fermentation times was conducted. Then, in the in vivo experiment, 10 sheep were divided into a control group which was fed a basal diet, and a 5-HTP group supplemented with 8 mg/kg 5-HTP for 60 days. The results showed that 5-HTP supplementation had a significant effect on in vitro DMD, pH, NH3-N, acetic acid, propionic acid, and TVFA concentrations. 5-HTP altered rumen bacteria composition and diversity indices including Chao1, Shannon, and Simpson. Moreover, the in vivo study on sheep confirmed that supplementing with 8 mg/kg of 5-HTP improved rumen fermentation efficiency and microbial composition. This led to enhanced sheep growth performance and increased involvement in the tryptophan metabolic pathway, suggesting potential benefits. Dietary 5-HTP (8 mg/kg DM) improves sheep growth performance by enhancing ruminal functions, antioxidant capacity, and tryptophan metabolism. This study can provide a foundation for the development of 5-HTP as a functional feed additive in ruminants' production.

Utilization of Free and Dipeptide-Bound Formyline and Pyrraline by Saccharomyces Yeasts.

The utilization of the glycated amino acids formyline and pyrraline as well as their peptide-bound derivatives by 14 Saccharomyces yeasts, including 6 beer yeasts (bottom and top fermenting), one wine yeast, 6 strains isolated from natural habitats and one laboratory reference yeast strain (wild type) was investigated. All yeasts were able to metabolize glycated amino acids via the Ehrlich pathway to the corresponding Ehrlich metabolites. While formyline and small amounts of pyrraline entered the yeast cells via passive diffusion, the amounts of dipeptide-bound MRPs, especially the dipeptides glycated at the C-terminus, decreased much faster, indicating an uptake into the yeast cells. Furthermore, the glycation-mediated hydrophobization in general leads to an faster degradation rate compared to the native lysine dipeptides. While the utilization of free formyline is yeast-specific, the amounts of (glycated) dipeptides decreased faster in the presence of brewer's yeasts, which also showed a higher formation rate of Ehrlich metabolites compared to naturally isolated strains. Due to rapid uptake of alanyl dipeptides, it can be assumed that the Ehrlich enzyme system of naturally isolated yeasts is overloaded and the intracellularly released MRP is primarily excreted from the cell. This indicates adaptation of technologically used yeasts to (glycated) dipeptides as a nitrogen source.

Development of a defined mixed starter culture for the improvement of Tej, Ethiopian honey wine.

Ethiopian honey wine, Tej, is the most popular traditionally fermented alcoholic beverage in Ethiopia. Owing to the spontaneous fermentation process, the final product is neither predictable nor consistent in quality. Attempts have not been made before to solve this problem. Thus, the aim of this study was to develop a potential mixed starter culture of yeast and lactic acid bacteria isolated from Tej samples for the production of Tej with consistent quality. One hundred seventy-seven lactic acid bacteria and 194 yeasts were isolated from 30 Tej samples collected from southwest Ethiopia. After sequentially testing the isolates towards physiological stress tolerance and desired metabolic products, 10 lactic acid bacteria and 10 yeast isolates were screened. Later, four lactic acid bacteria and four yeast isolates were found to be compatible in co-culture tests. Finally, the combination of lactic acid bacteria and yeast isolates was formulated using the design of expert version 7.0.0 software, and six formulates (F #1-6) were designed. Controlled Tej fermentation was performed under laboratory conditions using six lactic acid bacteria-yeast starter culture formulations. The sensory attributes, in terms of color, flavor, odor, turbidity, and overall acceptance analysis scored 4.8/5.0 (F #2) and 4.7/5.0 (F #6), with mean significant variations (p < 0.05) among the other formulates. These two formulates were considered the best-mixed starter cultures compared to the control and other formulates. Matrix-assisted laser desorption ionization-time of flight analysis revealed that the lactic acid bacteria starters (AAUL7 and AAUL10) belonged to Lactobacillus paracasei. While the yeast starters (AAUY2 and AAUY8) belonged to Saccharomyces cerevisiae. These mixed lactic acid bacteria-yeast starter cultures could be used as the best starter culture for the fermentation of Ethiopian honey wine, Tej, with consistent quality.

Investigating the concomitant production of carotenoids and lipids by the yeast Rhodosporidium babjevae using sugarcane bagasse hydrolysate or corn steep liquor

The yeast Rhodosporidium babjevae was used to produce carotenoids, lipids, and fatty acids using sugarcane bagasse hydrolysates or glucose. Investigating the impact of glucose concentrations (10–150 g/L), the yeast showed the capability to yield 16.9 g/L biomass and 10.5 g/L lipids at 60 g glucose/L, while the highest pigments concentration (312 μg/g) was observed at 10 g glucose/L. Pre-treatment methods, including organosolv (ethanol 60 %(w/w)), alkali (NaOH 2 %(w/v)) and alkaline-organosolv (ethanol 60 %(w/w) and NaOH 2 %(w/v)) were used on the sugarcane bagasse following enzymatic hydrolysis. Among these, alkaline organosolv pre-treatment exhibited the most effective lignin removal of 81.9 %, releasing 46.7 g/L sugars by hydrolysis. Yeast cultivation on the enzymatic hydrolysates of alkaline organosolv pre-treatment resulted in the highest biomass (9.6 g/L) and lipids (5.0 g/L) yields. However, the highest content of pigments (280 μg/g) was achieved when the yeast was cultivated on the enzymatic hydrolysates of organosolv pre-treatment. Production of 11.3 g/L biomass, 204 μg/g pigments, and 6.7 g/L lipids established that corn steep liquor holds potential as a cheap and viable alternative to yeast extract as nitrogen source. The analysis of fatty acid composition revealed the prevalence of oleic acid and linoleic acid as dominant components.