Cerevisiae Research Articles

Higher-order epistasis within Pol II trigger loop haplotypes.

RNA polymerase II (Pol II) has a highly conserved domain, the trigger loop (TL), that controls transcription fidelity and speed. We previously probed pairwise genetic interactions between residues within and surrounding the TL for the purpose of understand functional interactions between residues and to understand how individual mutants might alter TL function. We identified widespread incompatibility between TLs of different species when placed in the Saccharomyces cerevisiae Pol II context, indicating species-specific interactions between otherwise highly conserved TLs and its surroundings. These interactions represent epistasis between TL residues and the rest of Pol II. We sought to understand why certain TL sequences are incompatible with S. cerevisiae Pol II and to dissect the nature of genetic interactions within multiply substituted TLs as a window on higher order epistasis in this system. We identified both positive and negative higher-order residue interactions within example TL haplotypes. Intricate higher-order epistasis formed by TL residues was sometimes only apparent from analysis of intermediate genotypes, emphasizing complexity of epistatic interactions. Furthermore, we distinguished TL substitutions with distinct classes of epistatic patterns, suggesting specific TL residues that potentially influence TL evolution. Our examples of complex residue interactions suggest possible pathways for epistasis to facilitate Pol II evolution.

Genotypic characterization of β-lactamase resistant Salmonella species isolated from different quail houses and their susceptibility to extracts of thyme and chitosan, and Saccharomyces Cerevisiae

ABSTRACT This study aimed to identify Salmonella species in two systems for quail raising (deep litter system and cages system). Determination of antibiotic susceptibility, β-lactamase resistance genes profile and susceptibility to chitosan, thyme extract and S. cerevisiae was examined. Fifteen Salmonella isolates were isolated from eighty cloacal swabs examined, with a higher incidence from those of deep litter systems than cage housing systems. Regarding the environmental samples, Salmonella was higher in the deep litter system than in cage housing. Serological identification of 27 salmonella isolates typed them into 8 Salmonella spp. including S. Tsevie, S. Infantis, S. Kentucky, S. Typhimurium, S. Larochelle, S. Molade, S. Enteritidis and S. Labadi. The profile of β-lactamase-resistance genes revealed the presence of blaTEM and blaSHV genes in S. Typhimurium and S. Infantis; blaTEM and blaCTX-M1 genes in S. Enteritidis and S. Molade, while S. Kentucky and S. Tsevie were positive for the blaCTX-M1 gene only. S. Larochelle was negative for all of them. Thyme and chitosan extract showed that S. cerevisiae had the highest inhibitory activity against the tested strains. In conclusion, farms with superior biosecurity and hygiene practices will reduce the risk of pathogens and eliminate the need for prophylactic antibiotics, thereby preventing antibiotic resistance.

Not all cytochrome b5s are created equal: How a specific CytB5 boosts forskolin biosynthesis in Saccharomyces cerevisiae

Cytochrome B5s, or CytB5s, are small heme-binding proteins, ubiquitous across all kingdoms of life that serve mainly as electron donors to enzymes engaged in oxidative reactions. They often function as redox partners of the cytochrome P450s (CYPs), a superfamily of enzymes participating in multiple biochemical processes. In plants, CYPs catalyze key reactions in the biosynthesis of plant specialized metabolites with their activity dependent on electron donation often from cytochrome P450 oxidoreductases (CPRs or PORs). In eukaryotic microsomal CYPs, CytB5s frequently participate in the electron transfer process although their exact role remains understudied, especially in plant systems. In this study, we assess the role of CytB5s in the heterologous biotechnological production of plant specialized metabolites in yeast. For this, we used as a case-study the biosynthesis of forskolin - a bioactive diterpenoid produced exclusively from the plant Coleus forskohlii. The complete biosynthetic pathway for forskolin is known and includes three CYP enzymes. We reconstructed the entire forskolin pathway in the yeast Saccharomyces cerevisiae, and upon co-expression of the three CytB5s - identified in C. forskohlii transcriptomes - alleviation of a CYP-related bottleneck step was noticed only when a specific CytB5, CfCytB5A, was used. Co-expression of CfCytB5A in yeast, in combination with forskolin pathway engineering, resulted in forskolin production at titers of 1.81 g/L in a bioreactor. Our findings demonstrate that CytB5s not only play an important role in plant specialized metabolism but also, they can interact with precision with specific CYPs, indicating that the properties of CytB5s are far from understood. Moreover, our work highlights how CytB5s may act as indispensable components in the sustainable microbial production of plant metabolites, when their biosynthetic pathways involve CYP enzymes.

The Effect of Saccharomyces cerevisiae in Commercial Feed on Feed Utilization Efficiency, Growth Performance, and Blood Profile of Freshwater Pomfret (Colossoma macropomum) Fingerlings

The Effect of Saccharomyces cerevisiae in Commercial Feed on Feed Utilization Efficiency, Growth Performance, and Blood Profile of Freshwater Pomfret (Colossoma macropomum) Fingerlings

Fusarium graminearum Ste2 and Ste3 Receptors Undergo Peroxidase-Induced Heterodimerization when Expressed Heterologously in Saccharomyces cerevisiae.

Fusarium graminearum FgSte2 and FgSte3 are G-protein coupled receptors (GPCRs) shown to play roles in hyphal chemotropism and fungal plant pathogenesis in response to activity arising from host-secreted peroxidases. Here, we follow up on the observation that chemotropism is dependent on both FgSte2 and FgSte3 being present; testing the possibility that this might be due to formation of an FgSte2-FgSte3 heterodimer. Bioluminescence resonance energy transfer (BRET) analyses were conducted in Saccharomyces cerevisiae, where the addition of horse radish peroxidase (HRP) was found to increase the transfer of energy from the inducibly-expressed FgSte3-Nano luciferase donor, to the constitutively-expressed FgSte2-yellow fluorescent protein (YFP) acceptor, compared to controls. A partial response was also detected when an HRP-derived ligand-containing extract was enriched from F. graminearum spores and applied instead of HRP. In contrast, substitution with pheromones or an unrelated bovine GPCR, rhodopsin-YFP used as acceptor, eliminated all BRET responses. Interaction results were validated by affinity pulldown and receptor expression was validated by confocal immunofluorescence microscopy. Taken together these findings demonstrate the formation of HRP and HRP-derived ligand stimulated heterodimers between FgSte2 and FgSte3. Outcomes are discussed from the context of the roles of ligands and reactive oxygen species in GPCR dimerization.

Strain selection and adaptation of a fungal-yeast-microalgae consortium for sustainable bioethanol production and wastewater treatment from livestock wastewater

This study explores the potential of strain selection and adaptation for developing a fungi-yeast-microalgae consortium capable of integrated bioethanol production and livestock wastewater treatment. We employed a multi-stage approach involving isolation and strain selection/adaptation of these consortiums. The study started with screening some isolated fungi to grow on the cellulosic biomass of the livestock wastewater (saccharification) followed by a fermentation process using yeast for bioethanol production. The results revealed that Penicillium chrysogenum (Cla) and Saccharomyces cerevisiae (Sc) produced a remarkable 99.32 ppm of bioethanol and a concentration of glucose measuring 0.56 mg ml− 1. Following the impact of fungi and yeast, we diluted the livestock wastewater using distilled water and subsequently inoculated Nile River microalgae into the wastewater. The findings demonstrated that Chlorella vulgaris emerged as the dominant species in the microalgal community. Particularly, the growth rate reached its peak at a 5% organic load (0.105385), indicating that this concentration provided the most favorable conditions for the flourishing of microalgae. The results demonstrated the effectiveness of the microalgal treatment in removing the remaining nutrients and organic load, achieving a 92.5% reduction in ammonia, a 94.1% reduction in nitrate, and complete removal of phosphate (100%). The algal treatment also showed remarkable reductions in COD (96.5%) and BOD (96.1%). These findings underscore the potential of fungi, yeast, and Nile River microalgae in the growth and impact on livestock wastewater, with the additional benefit of bioethanol production.Graphical abstract

Evaluating the efficacy of probiotics and ascorbic acid as anti-stress agents against heat stress in broiler chickens.

Heat stress poses a substantial challenge to poultry production worldwide, highlighting the urgent need for effective management strategies. This study investigated the efficacy of probiotics (Saccharomyces cerevisiae) and ascorbic acid as antistress agents using cloacal and body surface temperatures (CT and BST) as heat stress biomarkers in broiler chickens. A total of 56 broiler chicks were used for the experiment and were divided into four distinct groups: control, probiotics (1 g/kg of feed), ascorbic acid (200 mg/kg of feed) and the combination of probiotics and ascorbic acid (1 g/kg and 200 mg/kg of feed, respectively). The study lasted 35 days; measurements were taken for ambient temperature (AT), CT, and BST. The ambient temperature in the pens consistently exceeded the thermoneutral zone (TNZ) established for broiler chickens. The CT values for broiler chickens in the probiotic group were significantly lower (p < 0.05) compared to the control group. Additionally, the BST values in the probiotic and probiotic + ascorbic acid groups were significantly lower (p < 0.05) than those in the control group. The findings suggest that incorporating probiotics, with or without ascorbic acid, can effectively reduce CT and BST values in broiler chickens thereby, enhancing thermoregulation when compared to the control group. This implies that using probiotics in poultry diets may enhance health and growth performance, potentially leading to better feed efficiency and reduced reliance on antibiotics. Implementing these dietary strategies could improve the productivity and welfare of broiler chickens in commercial settings.

Isolation and purification of lactic acid bacteria and yeasts based on a multi-channel magnetic flow device and rapid qualitative and quantitative detection

Rapid isolation and identification of lactic acid bacteria and yeasts during fermentation is of great significance for quality control and regulation of fermented foods. In this study, we prepared a multi-channel magnetic flow device for rapid separation and purification of lactic acid bacteria and yeast, and based on SERS spectrum, we made rapid qualitative and quantitative analysis of Lactobacillus plantarum, Lactococcus lactis and Saccharomyces cerevisiae. The results showed that the synthesized Synthesized Fe3O4-Van antibiotic magnetic beads are paramagnetic; Fe3O4-Van antibiotic magnetic beads achieved capture efficiencies of more than 98.5 % for both L. plantarum and L. lactis at 102–104 CFU/mL, respectively. Separation and purification efficiency of single S. cerevisiae, L. plantarum and L. lactis by multi-channel magnetic flow device all reached more than 98 % with good isolation and purification results. The SERS spectra of the three microorganisms were classified and analyzed using linear discriminant analysis (LDA), and the accuracy of the established LDA model was 100 %, which completely differentiated the SERS spectra of the three microorganisms,and realized the qualitative identification of L. plantarum, L. lactis, and S. cerevisiae, and finally, quantitative model was established with the logarithmic values (lg C) of different concentrations of L. plantarum, L. lactis, and S. cerevisiae as the horizontal coordinates, and the Raman intensities at their strongest characteristic peaks of 512 cm−1, 1669 cm−1, and 1125 cm−1, respectively, were used as vertical coordinates to establish a quantitative model, with the lowest detection limit of 10 CFU/mL, and the digital quantification of lactic acid bacteria and yeast were achieved. It provided an effective means for real-time monitoring and tracking of the dynamics of lactic acid bacteria and yeast in the fermentation process and the quality control of fermented foods.

Nutrient Consumption Patterns of Saccharomyces cerevisiae and Their Application in Fruit Wine Fermentation

This study aimed to evaluate the nutritional requirements of Saccharomyces cerevisiae to improve low ethanol production in some fruit wines. The growth kinetics, ethanol production and nutrient requirements of S. cerevisiae were analyzed in chemically defined media. The results revealed that Ca2⁺, Fe2⁺, Co2⁺, Mo2⁺, Cu2⁺ and BO₃3− were predominantly utilized during the late lag phase, whereas free amino acids, nicotinic acid, calcium pantothenate, Na⁺ and Mg2⁺ were mainly consumed during the logarithmic phase. Compared with the control medium, supplementation with threonine, inositol, calcium pantothenate, thiamine hydrochloride, riboflavin, biotin, MgSO4 or KH2PO4 significantly increased the ethanol content by 1.10-fold (p &lt; 0.05). Furthermore, adding key nutrients to noni-, guava- and mango juice significantly shortened the fermentation time and increased the final alcohol content of the fruit wines (p &lt; 0.05). This study provides scientific insights and effective methods for shortening fermentation time and increasing alcohol content with S. cerevisiae in some fruit wines.

Fermentative and Enological Features of Saccharomyces cerevisiae Populations Generated Through Adaptive Laboratory Evolution

Adaptive laboratory evolution (ALE) is a non-GMO technique utilized for the amelioration of wine yeast strains. Employing two-step ALE strategies, we recently acquired six evolved Saccharomyces cerevisiae populations with improved fermentative abilities compared to their parental strains in synthetic broths. Herein, we evaluated the qualities of the abovementioned evolved populations under real winemaking conditions, using the grape musts Assyrtiko and Roditis. The ethanol-tolerant populations evolved solely with glucose delayed to complete the fermentation due to slow fructose assimilation, albeit showing improved ethanol yields, compared to their parental strains. The volatile compounds of the evolved populations were significantly different from those of parental strains. Statistically significant differences were observed in the organoleptic profiles between the evolved populations’ and parental strains’ wines. Notably, wine from one evolved population (BLR200) was rated higher in overall aroma and quality. This study supports the magnitude of ALE strategies for the generation of novel wine yeasts.

Characterization of a High-Affinity Copper Transporter CTR1a in the White-Nose Syndrome Causing Fungal Pathogen Pseudogymnoascus destructans.

Copper is an essential micronutrient and the ability to scavenge tightly bound or trace levels of copper ions at the host-pathogen interface is vital for fungal proliferation in animal hosts. Recent studies suggest that trace metal ion acquisition is critical for the establishment and propagation of Pseudogymnoascus destructans, the fungal pathogen responsible for white-nose syndrome (WNS), on their bat host. However, little is known about these metal acquisition pathways in P. destructans. In this study, we report the characterization of the P. destructans high-affinity copper transporter VC83_00191 (PdCTR1a), which is implicated as a virulence factor associated with the WNS disease state. Using Saccharomyces cerevisiae as a recombinant expression host, we find that PdCTR1a can efficiently traffic Cu ions into the yeast cytoplasm. Complementary studies in the native P. destructans fungus provide evidence that PdCTR1a transcripts and protein levels are dictated by Cu-bioavailability in the growth media. Our study demonstrates that PdCTR1a is a functional high-affinity copper transporter and is relevant to Cu homeostasis pathways in P. destructans.

Intracellular dry mass density increases under growth-induced pressure

Cells that proliferate in confined environments develop mechanical compressive stress, referred to as growth-induced pressure, which inhibits growth and division across various organisms. Recent studies have shown that in these confined spaces, the diffusivity of intracellular nanoparticles decreases. However, the physical mechanisms behind this reduction remain unclear. In this study, we use quantitative phase imaging to measure the refractive index and dry mass density of Saccharomyces cerevisiae cells proliferating under confinement in a microfluidic bioreactor. Our results indicate that the observed decrease in diffusivity can be at least attributed to the intracellular accumulation of macromolecules. Furthermore, the linear scaling between cell content and growth-induced pressure suggests that the concentrations of macromolecules and osmolytes are maintained proportionally under such pressure in S. cerevisiae.

Engineering Saccharomyces cerevisiae for the Production of Punicic Acid-Rich Yeast Biomass.

Punicic acid (PuA), an unusual conjugated linolenic acid found in pomegranate, offers diverse health benefits and has potential applications in the food industry. Due to the limited availability of PuA from natural plant sources, there is growing interest in producing it through microbial fermentation. In this study, the yeast Saccharomyces cerevisiae, which is classified as "generally recognized as safe", was engineered to produce PuA using a results-driven approach. Genes potentially involved in PuA synthesis were integrated directly into the yeast genome, targeting Ty retrotransposon sites. Screening of the yeast transformants, followed by optimization of culture conditions, resulted in the production of 26.7% PuA within the yeast's total fatty acids. Further analysis revealed that the strain's triacylglycerol fraction contained over 22% PuA. By incorporating this health-promoting lipid into the nutritional profile of S. cerevisiae, the engineered strain could serve as a sustainable source of yeast biomass with enhanced nutritional value.

Optimization of Citrus Pulp Waste-Based Medium for Improved Bacterial Nanocellulose Production.

Bacterial nanocellulose (BC) has attracted significant attention across a wide array of applications due to its distinctive characteristics. Recently, there has been increasing interest in leveraging waste biomass to improve sustainability in BC biogenesis processes. This study focuses on optimizing the citrus pulp waste (CPW) medium to enhance BC production using Komagataeibacter sucrofermentans. The screening of initial medium pH, yeast extract, CPW sugar and inoculum concentrations was conducted using the Plackett-Burman design, with BC yield (mgDW/gCPW) as the model response. The significant parameters, i.e., CPW sugars and yeast extract concentrations, were optimized using response surface methodology, employing a five-level, two-factor central composite design. The optimized CPW-based growth medium resulted in a final yield of 66.7 ± 5.1 mgDW/gCPW, representing a 14-fold increase compared to non-optimized conditions (4.3 ± 0.4 mgBC/gCPW). Material characterization analysis indicated that the produced BC showed high thermal stability (30% mass retained at 600 °C) and a crystallinity index value of 71%. Additionally, to enhance process sustainability, spent baker's yeast hydrolysate (BYH) was assessed as a substitute for yeast extract, leading to a final BC titer of 9.3 ± 0.6 g/L.

Đánh giá hiệu quả ứng dụng bộ chế phẩm probiotic BIDI-AGRI và BIDI-AQUA trong nuôi tôm thẻ chân trắng thương phẩm

Whiteleg shrimp (Litopenaeus vannamei) farming is a major economic sector in many countries worldwide. Infectious diseases and environmental pollution in shrimp ponds have resulted in significant economic losses for the aquaculture industry. This study presented the effects of a BIDI-AGRI and BIDI-AQUA probiotic blend consisting of Bacillus subtilis, Bacillus tequilensis, Bacillus amilolyquefaciens, Bacillus licheniformis, Lactobacillus acidophilus, Saccharomyces cerevisiae in shrimp cultured water treatment and shrimp growth. The BIDI-AGRI and BIDI-AQUA were applied in three commercial whiteleg shrimp ponds with surface areas ranging from 700 to 1200 m² for 90 days. Results showed that compared to the control, BIDI-AGRI and BIDI-AQUA improved several water quality parameters, including H₂S, NH₃, and COD concentrations, controlled Vibrio spp. populations within acceptable limits and enhanced shrimp immunity. Furthermore, shrimp ponds treated with BIDI-AGRI and BIDI-AQUA exhibited higher growth rates and yields. Due to lower investment costs and higher revenue, shrimp ponds using BIDI-AGRI and BIDI-AQUA achieved approximately 1.6 times higher profits compared to ponds using commercially available probiotics.

Isolation, identification, and technological characteristics analysis of yeast strains from Pyracantha fortuneana fruits fermentation liquid.

Pyracantha fortuneana (P. fortuneana), as a medicinal and edible plant resource, is rich in nutrients. In order to screen the high quality yeast used in Firebone fruit wine, 12 strains of yeast were isolated and purified from P. fortuneana fermentation broth by traditional pure culture method. They were identified by molecular biology as Pichia kudriavzevii (P. kudriavzevii) and Saccharomyces cerevisiae (S. cerevisiae), respectively. Strain HJ-2 could grow normally at 30℃, alcohol content 15%, SO2 mass concentration 360mg/L, pH 3.2, sucrose mass concentration 400g/L and glucose mass concentration 250g/L. Strain HJ-6 could grow normally at 30℃, alcohol content 3%, SO2 concentration 360mg/L, pH 3.2, sucrose concentration 250g/L, glucose concentration 300g/L. Based on the technological characteristics of fruit wine, S. cerevisiae HJ-2 has the potential of brewing P. fortuneana fruit wine. P. kudriavzevii HJ-6 has a low tolerance to ethanol and is suitable for the production of fermented beverages such as low-alcohol wine or beer.

The Role of Keeving in Modulating Fermentation and the Flavour Profiles of Apple Brandy

Keeving is the removal of nutrients from apple musts due to their binding to pectin, resulting in a slower fermentation and spontaneous arrest. The aim of this study was to determine the effect of keeving on the chemical composition of fermented apple must and on the volatile profile and sensory analysis of apple brandies. We compared the application of keeving during spontaneous fermentation with fermentation carried out by Saccharomyces cerevisiae (SafSpirit HG-1). We evaluated the impact of adding different doses of calcium chloride on various parameters of fermented musts and distillates. Calcium chloride had a greater effect on the ethanol concentration, total extract, and fermentation efficiency than on the type of fermentation used. However, a different phenomenon was observed with respect to the volatiles. The concentration of most of the higher alcohols, acetaldehyde, dodecanal, and geranylaceton, decreased after spontaneous fermentation and increased during the fermentation carried out with Saccharomyces cerevisiae SafSpirit HG-1. In general, the application of keeving contributed to a decrease in the concentration of ethyl and methyl esters, but caused an increase in the concentration of all acetate esters and terpenoids. When the amount of nutrients in the environment is limited and starvation occurs, microorganisms use the available nutrients for basic metabolic processes that allow them to survive and limit the formation of side metabolites such as volatiles. However, most of the samples fermented after the faecal depletion achieved high scores for the floral, fruity, and “overall note” parameters in the sensory analysis. This means that this method, carried out with a properly selected yeast strain, could be feasible for the distilling industry.

New Plasmid Constructs for Production of Yeast Based Oral Vaccines

In order to avoid conventional vaccines, there is an urgent need to look for new ways of vaccine generation that can cut down the cost, time, and the need for special requirements during storage and distribution. The development of oral peptide/protein-based vaccines could be an ideal alternative. Five expression plasmids are designed in this work for the production of Saccharomyces cerevisiae-based oral vaccines. Therefore, in this work genetic cassette with different organizations of the required genetic components for efficient gene expression is constructed in the pYES2 plasmid. The construction of five new plasmids for surface display of heterologous recombinant proteins in S. cerevisiae would enable easy insertion of the gene of interest into an optimized genetic cassette consisting of a strong host promoter GAL1, HA-tag for detection of recombinant enzymes, GAS1 gene with/without binding sequence coded for GAS1 cell wall protein and signal sequence for directing the protein into the secretory pathway. After the construction of the genetic cassettes, E gene coded for envelope protein of Chikungunya and Dengue virus is inserted in the genetic cassettes. Genetic models pYES2HAGAS1EgeneChikungunya, pYES2HAGAS1 Egene Dengue, pYES2GAS1 Egene ChikungunyaHA, and pYES2GAS1EGeneDengueHA are constructed with different genetic organizations to display envelope protein of Chikungunya and Dengue virus in the cell surface of S. cerevisiae. In the pYES2SignalEGeneDengueHA construction, envelope protein will be secreted by S. cerevisiae into the media. The constructed vectors are specialized to regulate the expression and surface display of heterologous recombinant proteins in Saccharomyces cerevisiae and will be suitable for large-scale oral vaccine production.

The effect of indigenous non-Saccharomyces yeasts on the volatile profile of Maraština wine: Monoculture versus sequential fermentation

Non-Saccharomyces yeasts are presented as a new promising tool in winemaking to enhance aroma complexity in fermentation with Saccharomyces yeasts. Indigenous yeasts are recognized for their better adaptation to environmental conditions and for highlighting the unique terroir impact on wine aroma characteristics. To study the individual impact of ten indigenous non-Saccharomyces yeasts on the primary metabolites and volatile compounds of wine from autochthonous Croatian Maraština variety, Hyphopichia pseudoburtonii, Metschnikowia chrysoperlae, Metschnikowia sinensis/shanxiensis, Metschnikowia pulcherrima, Lachancea thermotolerans, Hanseniaspora uvarum, Hanseniaspora guilliermondii, Hanseniaspora pseudoguilliermondii, Pichia kluyveri, and Starmerella apicola were inoculated in sterile grape juice in monoculture fermentations. Additionally, seven of them were also studied in sequential fermentation of sterile grape juice with commercial Saccharomyces cerevisiae. A targeted approach was used for the identification of volatile compounds via headspace solid-phase microextraction coupled with gas chromatography and a mass spectrometer. P. kluyveri and H. uvarum produced higher total concentrations of terpenes in sequential versus monoculture fermentation. Sequential fermentation of M. chrysoperlae, L. thermotolerans, and P. kluyveri with S. cerevisiae resulted in higher production of C13-norisoprenoids. The esters concentration was higher in monocultures for L. thermotolerans, H. uvarum, and H. guilliermondii, whereas other isolates showed higher concentrations in sequential fermentations. The results highlighted different indigenous yeast metabolisms and provided promising insights into potential new non-Saccharomyces starter cultures as a first step in their selection, with several species characterized in terms of their potential effect on the aroma profile.

Automated yeast cultivation control using a biosensor and flow cytometry.

Effective microbial bioprocessing relies on maintaining ideal cultivation conditions, highlighting the necessity for tools that monitor and regulate cellular performance and robustness. This study evaluates a fed-batch cultivation control system based on at-line flow cytometry monitoring of intact yeast cells having a fluorescent transcription factor-based redox biosensor. Specifically, the biosensor assesses the response of an industrial xylose-fermenting Saccharomyces cerevisiae strain carrying the TRX2p-yEGFP biosensor for NADPH/NADP+ ratio imbalance when exposed to furfural. The developed control system successfully detected biosensor output and automatically adjusted furfural feed rate, ensuring physiological fitness at high furfural levels. Moreover, the single-cell measurements enabled the monitoring of subpopulation dynamics, enhancing control precision over traditional methods. The presented automated control system highlights the potential of combining biosensors and flow cytometry for robust microbial cultivations by leveraging intracellular properties as control inputs.