Strains Of Yeast Saccharomyces Cerevisiae Research Articles

Pre-adaptation of yeast (Saccharomyces cerevisiae) strains to very high gravity can improve fermentation parameters and reduce osmotic stress

Pre-adaptation of yeast (Saccharomyces cerevisiae) strains to very high gravity can improve fermentation parameters and reduce osmotic stress

Effects of ultrasound and microwave treatments on oenological yeast microbiota: comparison with a flash pasteurisation

SummaryThe control of yeast biota in wine could be a challenge, as different genera could coexist with different effects on wine quality; generally, SO2 plays an important role in this context, while other approaches have been proposed. In this article, two Microwave (MW) and two Ultrasound treatments (US) were tested and compared to a flash pasteurisation on seven yeast strains (Saccharomyces cerevisiae, Saccharomyces bayanus, Brettanomyces spp., Schizosaccharomyces pombe, Hanseniaspora osmophila, Zygosaccharomyces bailii, Pichia membranifaciens); viable count (viability) and yeast growth kinetic (vitality) were assessed. The treatments reduced viable count by 27%, with a higher biocidal effect of US, depending on yeast and treatment conditions; they also impaired yeast growth causing a delay, which is a significant increase of the lag phase, while the effect on the growth rate or the maximum OD attained in the steady state was less significant. In conclusion, the US generally exerted a stronger effect than MW and flash pasteurisation and Schizo. pombe and Z. bailii were the less resistant yeasts. A partial uncoupling was found between vitality and viability for Z. bailii, while in Schizo. pombe the delay in the growth was generally related to the decrease in the cell count immediately after the treatment.

Interrelations Between Probiotics, Gut Microbiota, Intestinal Barrier, and Immune Response; Focusing on Diarrhea in Dairy Calves1

The interplay between gut microbiota and host health has attracted significant interest in the animal science community. Maintaining gut microbiota homeostasis by supplementing probiotics to treat clinical conditions like calf diarrhea is an emerging area of research nowadays because of increased concerns regarding antimicrobial resistance (AMR) and drug residues in animal products. Probiotics reduce the incidence of calf diarrhea by increasing the gut microbiota diversity and richness with more commensal bacteria such as Lactobacillus and Bifidobacterium that produce antimicrobial compounds, as well as modulating the immune response by increasing cytokines, Interleukin-2 (IL-2), IL-4, IL-6, IL-10, and reducing tumor necrosis factor-α (TNF-α), by increasing production of antibodies, especially immunoglobulin E (IgE), also IgG, differentiating naïve Th lymphocytes (Tho) into Th1, hence stimulate innate immunity and prime the adaptive immune response. Specific probiotic strains of bacteria and yeast (Saccharomyces cerevisiae) derived probiotics maintain the integrity of the intestinal barrier. In this review, data is being organized to address the role of probiotics in treating calf diarrhea by modulating gut microbiota and stimulating an immune response against notorious pathogens, to present animal and veterinary scientists and nutritionists with a new concept to treat infectious diseases from the perspective of the gut microbiota, increasing animal health, performance, and welfare. In conclusion, health status and gut microbiome are strongly interlinked. Research data indicated a significant reduction in the incidence of diarrhea after probiotic administration. If interrelations between probiotics and existing gut microbiota are explored more quantitatively, novel antibiotic substitutes can emerge in the future.

Autochthonous Ingredients for Craft Beer Production

Innovation in the beer market focuses on research into the different ingredients that make up this popular and sought-after beverage. Some of these innovations have focused on the use of autochthonous ingredients, which bring added value to products, reduce production costs and environmental impact, and provide beers with special organoleptic characteristics. This study focuses on the use of autochthonous ingredients from the Community of Madrid and the design of four recipes that are suited to these ingredients and consumer demand. In terms of the ingredients used, the micromalting of barley grown in the CM was carried out, while hops (Cascade variety for bittering hops and wild hops Rayo verde Tajuña and Torreta Tajuña for aroma) and yeast strains (Saccharomyces cerevisiae G 520 and G 354) were selected from previous studies, which were also sourced from the CM. For the barley malt, parameters such as protein content were determined, which was at the maximum limit but did not affect the final quality of the beers. The content of α- and β-acids in the hops was also determined, being higher in the Cascade variety. Different physicochemical analyses (GC, aromatic compounds, bitterness, colour, SO2, polyphenols, and antioxidant capacity) were carried out to determine the quality of the beers obtained from the four recipes designed, as well as tastings by a panel of experts and consumers. Finally, two recipes were rated the best by a panel of experts and consumers, where the cereal, hop, and fruit flavours stood out for their aromatic profile.

Kinetic Study of Fig Syrup Fermentation by Genetically Modified Saccharomyces cerevisiae Yeast Strains: A Physicochemical Approach to the Yeast Strain Life Cycle

Reversed-flow gas chromatography (R.F.G.C.) was employed to assess the impact of genetic modification on Saccharomyces cerevisiae yeast strains during the process of alcoholic fermentation, utilizing fig syrup. Multiple fermentations were carried out at various temperatures to evaluate the influence of genetic modifications on yeast strain efficiency. The study involved a wild-type yeast strain, W303, as a control and two genetically modified strains, W_M4_533 and W_M4_558, sharing the same genetic background as the wild type. Notably, the genetic modifications in the Msn4p transcription factor involved the substitution of serine residues with alanine at positions 533 and 558, resulting in the development of psychrophilic or ethanol-resistant strains. Utilizing the R.F.G.C. method enabled the differentiation of the duration of alcoholic fermentation phases, providing insights correlated to the yeast cell life cycle. The values of rate constants (k) for each phase, conducted with both wild-type and genetically modified cells using RFGC, aligned with the existing literature. Additionally, the calculation of activation energies for distinct phases revealed lower values for genetically modified strains compared to wild-type strains. This decrease in activation energies suggests enhanced efficiency in the alcoholic fermentation process for the genetically modified strains.

Specific Antimicrobial Activities Revealed by Comparative Evaluation of Selected Gemmotherapy Extracts.

Nowadays, unprecedented health challenges are urging novel solutions to address antimicrobial resistance as multidrug-resistant strains of bacteria, yeasts and moulds are emerging. Such microorganisms can cause food and feed spoilage, food poisoning and even more severe diseases, resulting in human death. In order to overcome this phenomenon, it is essential to identify novel antimicrobials that are naturally occurring, biologically effective and increasingly safe for human use. The development of gemmotherapy extracts (GTEs) using plant parts such as buds and young shoots has emerged as a novel approach to treat/prevent human conditions due to their associated antidiabetic, anti-inflammatory and/or antimicrobial properties that all require careful evaluations. Seven GTEs obtained from plant species like the olive (Olea europaea L.), almond (Prunus amygdalus L.), black mulberry (Morus nigra L.), walnut (Juglans regia L.), blackberry (Rubus fruticosus L.), blackcurrant (Ribes nigrum L.) and bilberry (Vaccinium myrtillus L.) were tested for their antimicrobial efficiency via agar diffusion and microbroth dilution methods. The antimicrobial activity was assessed for eight bacterial (Bacillus cereus, Staphylococcus aureus, Salmonella enterica subsp. enterica, Proteus vulgaris, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Listeria monocytogenes), five moulds (Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus, Penicillium citrinum, Penicillium expansum) and one yeast strain (Saccharomyces cerevisiae). The agar diffusion method revealed the blackberry GTE as the most effective since it inhibited the growth of three bacterial, four moulds and one yeast species, having considered the total number of affected microorganism species. Next to the blackberry, the olive GTE appeared to be the second most efficient, suppressing five bacterial strains but no moulds or yeasts. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined for each GTE and the microorganisms tested. Noticeably, the olive GTE appeared to feature the strongest bacteriostatic and bactericidal outcome, displaying specificity for S. aureus, E. faecalis and L. monocytogenes. The other GTEs, such as blueberry, walnut, black mulberry and almond (the list indicates relative strength), were more effective at suppressing microbial growth than inducing microbial death. However, some species specificities were also evident, while the blackcurrant GTE had no significant antimicrobial activity. Having seen the antimicrobial properties of the analysed GTEs, especially the olive and black mulberry GTEs, these could be envisioned as potential antimicrobials that might enhance antibiotic therapies efficiency, while the blackberry GTE would act as an antifungal agent. Some of the GTE mixtures analysed have shown interesting antimicrobial synergies, and all the antimicrobial effects observed argue for extending these studies to include pathological microorganisms.

Expression of chitosanase from Aspergillus fumigatus chitosanase in Saccharomyces cerevisiae by CRISPR-Cas9 tools.

Chitooligosaccharides (COS) find numerous applications due to their exceptional properties. Enzymatic hydrolysis of chitosan by chitosanase is considered an advantageous route for COS production. Heterologous expression of chitosanase holds significant promise, yet studies using commonly employed Escherichia coli and Pichia pastoris strains encounter challenges in subsequent handling and industrial scalability. In this investigation, we opted for using the safe yeast strain Saccharomyces cerevisiae (GRAS), obviating the need for methanol induction, resulting in successful expression. Ultimately, utilizing the GTR-CRISPR editing system, shake flask enzyme activity reached 2 U/ml. The optimal chitosanase activity was achieved at 55℃ and pH 5, with favorable stability between 30 and 50°C. Following a 2-h catalytic reaction, the product primarily consisted of chitobiose to chitotetraose, predominantly at the chitotriose position, with a slight increase in chitobiose content observed during the later stages of enzymatic hydrolysis. The results affirm the feasibility of heterologous chitosanase expression through Saccharomyces cerevisiae, underscoring its significant industrial potential.

Isolation and identification of dominant lactic acid bacteria and yeast species from teff (Eragrostis teff) injera dough fermentation

ABSTRACT Traditional fermentation using natural microorganisms often results in uncontrolled and inconsistent product quality. Identifying dominant LAB and yeast species through morphological, physiological, and biochemical characterization, this study aimed to optimize the traditional fermentation process of injera dough fermentation. Ersho and dough were prepared in the laboratory, and LAB and yeast were identified and enumerated by smearing on a petri dish at 24-hour intervals from 0 to 96 hours of fermentation. The growth kinetics of both LAB and yeast in fermented teff dough significantly increased (p < 0.05) from 8.91 to 9.97 log CFU/gm and 5.98 to 7.62 log CFU/gm as fermentation time increased. Five dominant LAB species (Lactobacillus fermentum, Lactobacillus Brevis, Lactobacillus Plantarum, Bacillus subtilis, and Enterococcus casseliflavus) and three dominant yeast strains (Saccharomyces cerevisiae, Candida krusei, and Pichia kudriavzevii) were identified. Based on the physicochemical analysis and sensory acceptability result, application of a single starter culture in teff dough fermentation improved fermentation time and acid production.

Chloroplast phosphate transporter CrPHT4-7 regulates phosphate homeostasis and photosynthesis in Chlamydomonas.

In eukaryotic cells, phosphorus is assimilated and utilized primarily as phosphate (Pi). Pi homeostasis is mediated by transporters that have not yet been adequately characterized in green algae. This study reports on PHOSPHATE TRANSPORTER 4-7 (CrPHT4-7) from Chlamydomonas reinhardtii, a member of the PHT4 transporter family, which exhibits remarkable similarity to AtPHT4;4 from Arabidopsis (Arabidopsis thaliana), a chloroplastic ascorbate transporter. Using fluorescent protein tagging, we show that CrPHT4-7 resides in the chloroplast envelope membrane. Crpht4-7 mutants, generated by the CRISPR/Cas12a-mediated single-strand templated repair, show retarded growth, especially in high light, reduced ATP level, strong ascorbate accumulation, and diminished non-photochemical quenching in high light. On the other hand, total cellular phosphorous content was unaffected, and the phenotype of the Crpht4-7 mutants could not be alleviated by ample Pi supply. CrPHT4-7-overexpressing lines exhibit enhanced biomass accumulation under high light conditions in comparison with the wild-type strain. Expressing CrPHT4-7 in a yeast (Saccharomyces cerevisiae) strain lacking Pi transporters substantially recovered its slow growth phenotype, demonstrating that CrPHT4-7 transports Pi. Even though CrPHT4-7 shows a high degree of similarity to AtPHT4;4, it does not display any substantial ascorbate transport activity in yeast or intact algal cells. Thus, the results demonstrate that CrPHT4-7 functions as a chloroplastic Pi transporter essential for maintaining Pi homeostasis and photosynthesis in C. reinhardtii.

Fermentation performance of Saccharomyces cerevisiae yeast strain with high alcoholigenous power

Fermentation performance of Saccharomyces cerevisiae yeast strain with high alcoholigenous power

Lymphatic Transport and Immune Activation Effect by Locally Administered Extracellular Vesicles from Saccharomyces cerevisiae as Biocompatible Vaccine Adjuvants.

The yeast strain Saccharomyces cerevisiae is an eukaryotic organism that has been widely used for the production of fermented foods. Most cells secrete extracellular vesicles (EVs), small particles composed of lipid membranes. Elucidating the role of EVs as a new intercellular communication system and developing novel EV-based therapies have attracted the increased attention of researchers. Although recent studies have reported the secretion of EVs from S. cerevisiae, their in vivo fate and subsequent EV-mediated biological responses in the host are unclear. In this study, we characterized both the biodistribution of locally (intradermally and subcutaneously) administered Saccharomyces cerevisiae-derived EVs (S-EVs) and the EV-mediated immune responses to evaluate their potential use as biocompatible vaccine adjuvants. S-EVs were round but heterogeneous in size and contained glucan, DNA, and RNA. Their mean particle sizes and zeta potentials were approximately 177.5 nm and -14.6 mV, respectively. We provided evidence that locally administered S-EVs were delivered to the lymph nodes, mainly reaching the B-cell zone. Measurement of host immune reactions revealed that administration of S-EVs increased the expression of cytokine (tumor necrosis factor (TNF)-α) and costimulatory molecules (CD40, CD80, CD86), which are indicators of immune activation. Especially, subcutaneously injected S-EVs showed potent adjuvanticity, indicating that subcutaneous administration of S-EVs is the desirable approach for achieving effective immune stimulation. These findings will facilitate the development of novel EV-based immunotherapies.

Elucidation of the mescaline biosynthetic pathway in peyote (Lophophora williamsii).

Peyote (Lophophora williamsii) is an entheogenic and medicinal cactus native to the Chihuahuan desert. The psychoactive and hallucinogenic properties of peyote are principally attributed to the phenethylamine derivative mescaline. Despite the isolation of mescaline from peyote over 120 years ago, the biosynthetic pathway in the plant has remained undiscovered. Here, we use a transcriptomics and homology-guided gene discovery strategy to elucidate a near-complete biosynthetic pathway from l-tyrosine to mescaline. We identified a cytochrome P450 that catalyzes the 3-hydroxylation of l-tyrosine to l-DOPA, a tyrosine/DOPA decarboxylase yielding dopamine, and four substrate-specific and regiospecific substituted phenethylamine O-methyltransferases. Biochemical assays with recombinant enzymes or functional analyses performed by feeding putative precursors to engineered yeast (Saccharomyces cerevisiae) strains expressing candidate peyote biosynthetic genes were used to determine substrate specificity, which served as the basis for pathway elucidation. Additionally, an N-methyltransferase displaying broad substrate specificity and leading to the production of N-methylated phenethylamine derivatives was identified, which could also function as an early step in the biosynthesis of tetrahydroisoquinoline alkaloids in peyote.

Liquid submerged fermentation by selected microbial strains for onion skins valorization and its effects on polyphenols

Onion skins, actually recycled as organic fertilizers, could be used as a substrate in environmental-friendly bioprocesses to recover high-value bioactive compounds and food ingredients.In this work, a bioprospecting method was carried out including 94 bacterial and 45 yeast strains from several agri-food and environmental niches to verify their ability to grow on onion skins as unique nutrients source.Red and yellow onion skins were assessed by newly selected starter-driven liquid submerged fermentation assays mainly aimed at the release and modification of polyphenols through microbial activities. Fermented onion skins were also investigated as a inexpensive favourable source of microbial enzymes (amylases, proteases, lipases, esterases, cellulases, xylanases).In red onion skins, the treatment with Lactiplantibacillus plantarum TB 11–32 produced a slight increase of bioactive compounds in terms of total phenolics, whereas with the yeast strain Zygosaccharomyces mrakii CL 30 − 29 the quercetin aglycone content increased of about 25% of the initial raw material.In yellow onion skins inoculated, the highest content of phenolic compounds was detected with the yeast strain Saccharomyces cerevisiae En SC, while quercetin aglycone increased of about 60% of the initial raw material in presence of the bacterial strain L. plantarum C 180 − 34.In conclusion, the proposed microbial pre-treatment method can be a potential strategy to re-use onion skins as a fermentation substrate, and as a first step in the development of a biorefinery process to produce value-added products from onion by-products.

Diversity of Indigenous Saccharomyces cerevisiae Yeast Strains in Cabernet Sauvignon Fermentations from Utiel-Requena Region (Spain) as a Resource to Improve Wine Distinctiveness

The aim of this study was to isolate and characterize autochthonous Saccharomyces cerevisiae yeasts from Cabernet Sauvignon grape must and to analyze how they impacted the final characteristics of the wines obtained from the same type of must. This work was carried out in a “Pago” winery located in Requena, Spain. Twenty-two isolates were obtained from grape must. After molecular identification and typing, 11 different strains were determined. Growth-related parameters and metabolic characteristics (glucose and fructose consumption and ethanol, glycerol and acetic acid production) were determined via laboratory-scale fermentations (50 mL) of Cabernet Sauvignon must. From 2 L fermentation batches, the residual sugar, density, ethanol concentration, titratable and volatile acidities, pH and polyphenolic and volatile compounds were determined, as well as the sensory attributes of the produced wines. Differences between strains in the growth and metabolic parameters were observed. Eleven S. cerevisiae strain profiles were characterized to establish which strain or strains of the selected yeasts would produce the highest polyphenolic and aromatic concentrations of the red Cabernet wines made in the “Pago winery”, to then multiply them for their use to make the red wines of this winery.

Probiotic potential of a yeast strain Saccharomyces cerevisiae BGY12 isolated from an ethnic beverage of Jaunsar-Bawar region, Uttarakhand, India

Probiotic potential of a yeast strain Saccharomyces cerevisiae BGY12 isolated from an ethnic beverage of Jaunsar-Bawar region, Uttarakhand, India

Probiotic potential of Saccharomyces cerevisiae GILA with alleviating intestinal inflammation in a dextran sulfate sodium induced colitis mouse model

Recently, several probiotic products have been developed; however, most probiotic applications focused on prokaryotic bacteria whereas eukaryotic probiotics have received little attention. Saccharomyces cerevisiae yeast strains are eukaryotes notable for their fermentation and functional food applications. The present study investigated the novel yeast strains isolated from Korean fermented beverages and examined their potential probiotic characteristics. We investigated seven strains among 100 isolates with probiotic characteristics further. The strains have capabilities such as auto-aggregation tendency, co-aggregation with a pathogen, hydrophobicity with n-hexadecane,1,1-diphenyl-2-picrylhydrazyl scavenging effect, survival in simulated gastrointestinal tract conditions and the adhesion ability of the strains to the Caco-2 cells. Furthermore, all the strains contained high cell wall glucan content, a polysaccharide with immunological effects. Internal transcribed spacer sequencing identified the Saccharomyces strains selected in the present study as probiotics. To examine the effects of alleviating inflammation in cells, nitric oxide generation in raw 264.7 cells with S. cerevisiae showed that S. cerevisiae GILA could be a potential probiotic strain able to alleviate inflammation. Three probiotics of S. cerevisiae GILA strains were chosen by in vivo screening with a dextran sulfate sodium-induced colitis murine model. In particular, GILA 118 down-regulates neutrophil–lymphocyte ratio and myeloperoxidase in mice treated with DSS. The expression levels of genes encoding tight junction proteins in the colon were upregulated, cytokine interleukin-10 was significantly increased, and tumor necrosis factor-α was reduced in the serum.

The Influence of Grape Clone and Yeast Strain on Varietal Thiol Concentrations and Sensory Properties of Graševina Wines.

Varietal thiol concentration in wine is influenced by numerous factors, of which grape variety and winemaking practices are often highlighted as the most important. Therefore, the aim of this work was to study the effects of grape clone and yeast strain (Saccharomyces and non-Saccharomyces) on the varietal thiols concentrations and sensory characteristics of Graševina (Vitis vinifera L.) white wines. Two grape clones were evaluated (OB-412 and OB-445) along with three different commercial yeast strains (Saccharomyces cerevisiae Lalvin Sensy and Sauvy, and Metschnikowia pulcherrima Flavia). The results showed that the concentration of varietal thiols in Graševina wines amounted up to a total of 226 ng/L. The clone OB-412 was characterized by its significantly higher concentrations, especially of 3-sulfanylhexanol (3SH) and 3-sulfanylhexyl acetate (3SHA). Moreover, alcoholic fermentation with pure S. cerevisiae Sauvy yeasts generally resulted in higher thiol concentrations, while sequential fermentation involving M. pulcherrima positively affected only the 4-methyl-4-sulfanyl-pentan-2-one (4MSP) concentration. Finally, sensory analysis showed that fermentation with pure S. cerevisiae Sauvy yeast also produced more desirable wines. The results suggest that clonal, and especially yeast strain, selections are important modulators of the aroma and sensory properties of wine.

Effects of a Blend of Live Yeast and Organic Minerals as an Alternative to Monensin on Intake, Digestibility, Performance and Beef Quality of Nellore Bulls Finished on Pasture with High Concentrate Supplementation

Effects of a blend of live yeast and organic minerals as an alternative to monensin and inorganic trace minerals for beef cattle finished on pasture with high concentrate supplementation, on growth performance, intake, digestibility, liver trace mineral and carcass characteristics were evaluated. Forty-eight Nellore bulls were blocked by initial body weight and randomly assigned to one of the two experimental diets. The animals were raised in an experimental pasture divided into 12 paddocks equipped with an electronic system for monitoring individual feeding behavior and feed intake. Treatments were: (1) Monensin (MON), 30 mg/kg supplement dry matter of sodium monensin and trace minerals supplementation from inorganic sources; (2) AdvantageTM (ADV), 1.6 g/kg supplement DM of a blend of live yeast (Saccharomyces cerevisiae strains) and organic trace minerals. The use of ADV instead of MON led to greater supplement intake and greater intake of dietary components. Bulls fed ADV also had higher digestibility of dry matter, organic matter, crude protein and non-fiber carbohydrates. Bulls fed MON had a greater number of visits to the feeder, however with a shorter time spent feeding per visit. The use of ADV resulted in higher average daily weight gain, and feed efficiency was similar between treatments. In the carcass, ADV tended toward greater Longissimus muscle area. Liver Zn concentration tended to be lower in the ADV treatment. The use of ADV generated higher meat lightness and redness. In summary, the blend of live yeast and organic minerals can be an alternative to monensin and inorganic sources of trace minerals for beef cattle finished on pasture with high concentrate supplementation, without negative effects on supplement feed efficiency and with benefits to animal growth.

Volatile compounds for biotechnological applications produced during competitive interactions between yeasts and fungi.

Fungi, yeasts and bacteria produce volatile compounds during their metabolism. In this study, the volatile compounds produced by yeast strains (Saccharomyces cerevisiae and Rhodotorula mucilaginosa) and fungal strains (Aspergillus carbonarius and Aspergillus ochraceus) during competitive interactions were investigated by solid-phase microextraction coupled with gas chromatography-mass spectrometry. Fifty-six volatile compounds were identified representing alcohols, aldehydes, esters, ketones, aromatic compounds, acids, furans, phenols, and nitrogen compounds, being the largest amount in the class of esters and alcohols. Eight compounds were identified only in interactive culture conditions such as 2-amino-1-propanol, isopropylamine, dimethylamine, pentyl propanoate, ethyl-2-aminopropanoate, acetone, oxalic acid, and β-elemene and five of these were produced in cocultures including A. carbonarius. These will be developed for future biotechnological applications such as in the pharmaceutical and biological industry to produce drugs. Antimicrobial and antifungal activities; Solvent and herbicide; flavoring ingredient; solvent, plastic synthesis, nail polish remover and thinner, pesticide and herbicide; important in the complexation of minerals in the soil; and plant-environment interactions, defending predators, pathogens, and competitors.

Characterization and Biological Studies of Synthesized Titanium Dioxide Nanoparticles from Leaf Extract of Juniperus phoenicea (L.) Growing in Taif Region, Saudi Arabia

Green synthesis of metal nanoparticles in nanosized form has acquired great interest in the area of nanomedicine as an environmentally friendly and cost-effective alternative compared to other chemical and physical methods. This study deals with the eco-friendly green synthesis of titanium dioxide nanoparticles (TiO2 NPs) utilizing Juniperus phoenicea leaf extract and their characterization. The biosynthesis of TiO2 NPs was completed in 3 h and confirmed by UV-Vis spectroscopy, a strong band at 205.4 nm distinctly revealed the formation of NPs. Transmissions electron microscopy (TEM) analysis showed the synthesized TiO2 NPs are spherical in shape, with a diameter in a range of 10–30 nm. The XRD major peak at 27.1° congruent with the (110) lattice plane of tetragonal rutile TiO2 phase. Dynamic light scattering (DLS) analysis revealed synthesized TiO2 NPs average particle size (hydrodynamic diameter) of (74.8 ± 0.649) nm. Fourier transmission infrared (FTIR) revealed the bioactive components present in the leaf extract, which act as reducing and capping agents. The antimicrobial efficacy of synthesized TiO2NPs against, Staphylococcus aureus, and Bacillus subtilis (Gram-positive), Escherichia coli and Klebsiella pneumoniae (Gram-negative), Yeast strain (Saccharomyces cerevisiae) and fungi (Aspergillus niger, and Penicillium digitatum) assayed by a disc diffusion method. TiO2NPs inhibited all tested strains by mean inhibition zone (MIZ), which ranged from the lowest 15.7 ± 0.45 mm against K. pneumoniae to the highest 30.3 ± 0.25 against Aspergillus niger. The lowest minimum inhibitory concentration (MIC) and bactericidal (MBC) values were 20 μL/mL and 40 μL/mL of TiO2NPs were observed against Asp. niger. Moreover, it showed significant inhibitory activity against human ovarian adenocarcinoma cells with IC50 = 50.13 ± 1.65 µg/mL. The findings concluded that biosynthesized TiO2 NPs using Juniperus phoenicea leaf extract can be used in medicine as curative agents according to their in vitro antibacterial, antifungal, and cytotoxic activities.