Search Results (2,959)

Late-spring frost events severely damage low-chill peach blossoms, causing significant yield losses. Although 5-aminolevulinic acid (ALA) enhances cold tolerance through the PpC3H37-PpWRKY18 module, the regulatory mechanism of ALA on PpC3H37 remains to be elucidated. Using yeast one-hybrid screening with the PpC3H37 promoter as bait, we identified PpDof9 as a key interacting transcription factor. A genome-wide analysis revealed 25 PpDof genes in peaches (Prunus persica). These genes exhibited variable physicochemical properties, with most proteins predicted as nuclear-localized. Subcellular localization experiments in tobacco revealed that PpDof9 was localized to the nucleus, consistent with predictions. A synteny analysis indicated nine segmental duplication pairs and tandem duplications on chromosomes 5 and 6, suggesting duplication events drove family expansion. A conserved motif analysis confirmed universal presence of the Dof domain (Motif 1). Promoter cis-element screening identified low-temperature responsive (LTR) elements in 12 PpDofs, including PpDof1, PpDof8, PpDof9, and PpDof25. The quantitative real-time PCR (qRT-PCR) results showed that PpDof1, PpDof8, PpDof9, PpDof15, PpDof16, and PpDof25 were significantly upregulated under low-temperature stress, and this upregulation was further enhanced by ALA pretreatment. Our findings demonstrate ALA-mediated modulation of specific PpDof TFs in cold response and provide candidates (PpDof1, PpDof9, PpDof8, PpDof25) for enhancing floral frost tolerance in peaches. Full article

The expression of recombinant proteins in heterologous hosts is a common strategy to obtain larger quantities of the “protein of interest” (POI) for scientific, therapeutic or commercial purposes. However, the experimental success of such an approach critically depends on the choice of an appropriate host system to obtain biologically active forms of the POI. The correct folding of the molecule, mediated by disulfide bond formation, is one of the most critical steps in that process. Here we describe the recombinant expression of hirudin, a leech-derived anticoagulant and thrombin inhibitor, in the yeast Komagataella phaffii (formerly known and mentioned throughout this publication as Pichia pastoris) and in two different strains of Escherichia coli, one of them being especially designed for improved disulfide bond formation through expression of a protein disulfide isomerase. Cultivation of the heterologous hosts and expression of hirudin were performed at different temperatures, ranging from 22 to 42 °C for the bacterial strains and from 20 to 30 °C for the yeast strain, respectively. The thrombin-inhibitory potencies of all hirudin preparations were determined using the thrombin time coagulation assay. To our surprise, the hirudin preparations of P. pastoris were considerably less potent as thrombin inhibitors than the respective preparations of both E. coli strains, indicating that a eukaryotic background is not per se a better choice for the expression of a biologically active eukaryotic protein. The hirudin preparations of both E. coli strains exhibited comparable high thrombin-inhibitory potencies when the strains were cultivated at their respective optimal temperatures, whereas lower or higher cultivation temperatures reduced the inhibitory potencies. Full article

Reducing post-harvest losses of cereal crops is a key challenge for ensuring global food security amid the limited arable land and growing population. This study investigates the effectiveness of electron beam irradiation (5 MeV, ILU-10 accelerator) as a physical decontamination method for various cereal crops cultivated in Kazakhstan. Samples were irradiated at doses ranging from 1 to 5 kGy, and microbiological indicators—including Quantity of Mesophilic Aerobic and Facultative Anaerobic Microorganisms (QMAFAnM), yeasts, and molds—were quantified according to national standards. Experimental results demonstrated an exponential decline in microbial contamination, with a >99% reduction achieved at doses of 4–5 kGy. The modeled inactivation kinetics showed strong agreement with the experimental data: R² = 0.995 for QMAFAnM and R² = 0.948 for mold, confirming the reliability of the exponential decay models. Additionally, key quality parameters—including protein content, moisture, and gluten—were evaluated post-irradiation. The results showed that protein levels remained largely stable across all doses, while slight but statistically insignificant fluctuations were observed in moisture and gluten contents. Principal component analysis and scatterplot matrix visualization confirmed clustering patterns related to radiation dose and crop type. The findings substantiate the feasibility of electron beam treatment as a scalable and safe technology for improving the microbiological quality and storage stability of cereal crops. Full article

Sustainability represents a significant global challenge, requiring a balance between environmental impact and the use of natural resources. White biotechnology, which uses microorganisms and enzymes for environmentally friendly products and processes, offers promising solutions to support a growing population. Within this context, the yeast Yarrowia lipolytica stands out, so we investigated the generation of biomass from two wild strains (ATCC 9773 and NRRL Y-50997) using different carbon sources. Additionally, protein content and amino acid profiles were assessed via standardized analytical methods to evaluate their potential as nutritional yeasts. Both strains demonstrated potential as nutritional yeasts, with biomass productivities of up to 35.5 g/L and 42 g/L, respectively. The protein content was high, with 58.8% for ATCC 9773 and 58.2% for NRRL Y-50997. Furthermore, the strains presented essential amino acid contents of 62.6% and 41.5%, with lysine being the most abundant amino acid. These findings underscore the versatility and productivity of Y. lipolytica, highlighting its potential for sustainable biotechnological applications such as single-cell protein production. Full article

Candidozyma auris is a multidrug-resistant, thermo- and osmotolerant yeast capable of persisting on biotic and abiotic surfaces, attributes likely linked to its cell wall composition. Here, seven putative genes encoding yapsins, aspartyl proteases GPI-anchored to the membrane or cell wall, were identified in the genomes of C. auris CJ97 and 20-1498, from clades III and IV, respectively. The C. auris YPS1 gene is orthologous to the SAP9 of C. albicans. The YPS7 gene is orthologous to YPS7 in C. glabrata and S. cerevisiae, so that they may share similar roles. An in silico analysis suggested an interaction between pepstatin and the catalytic domain of Yps1 and Yps7. Although this inhibitor, when combined with caffeine, had a subtle effect on the growth of C. auris, it induced alterations in the cell wall. CauYPS1 and CauYPS7 expression increased under nutrient starvation and NaCl, and at 42 °C. The transcriptome of the 20-1498 strain suggests that autophagy may play a role in thermal stress, probably degrading deleterious proteins or maintaining cell wall and vacuolar homeostasis. Therefore, CauYps1 and CauYps7 may play a role in the cell wall integrity of C. auris in stress conditions, and they could be a target of new antifungal or antivirulence agents. Full article

To explore an industrial fermentation approach for traditional mung bean sour pulp, this study isolated core microorganisms including lactic acid bacteria and yeasts from naturally fermented samples and constructed a synthetic microbial community. The optimized community consisted of Lactiplantibacillus pentosus, Lactococcus garvieae, and Cyberlindnera jadinii at a ratio of 7:3:0.1 and was used to ferment cooked mung bean pulp with a material-to-water ratio of 1:8 and 1% sucrose addition. Under these conditions, the final product exhibited significantly higher levels of protein (4.55 mg/mL), flavonoids (0.10 mg/mL), polyphenols (0.11 mg/mL), and vitamin C (7.75 μg/mL) than traditionally fermented mung bean sour pulp, along with enhanced antioxidant activity. The analysis of organic acids, free amino acids, and volatile compounds showed that lactic acid was the main acid component, the bitter amino acid content was reduced, the volatile flavor compounds were more abundant, and the level of harmful compound dimethyl sulfide was significantly decreased. These results indicate that fermentation using a synthetic microbial community effectively improved the nutritional quality, flavor, and safety of mung bean sour pulp. Full article

Pardosa pseudoannulata plays an important role in the biological control of insect pests. The inclusion of yeast in the culture medium is very important for the growth, development, and reproduction of Drosophila melanogaster, but there have been few studies on the influence of nutrients in the culture medium on spider development. In order to explore the effects of different yeast treatments on the growth and development of D. melanogaster and as a predator, P.  pseudoannulata, three treatments (no yeast, active yeast added, and inactivated yeast added) were adopted to modify the conventional D. melanogaster culture medium. The addition of yeast to the medium shortened the development time from larva to pupation in D. melanogaster. The emergence and larval developmental times of D. melanogaster reared with activated yeast were shorter than those of the group without yeast addition, which promoted D. melanogaster emergence and increased body weight. The addition of yeast to the medium increased the fat, protein, and glucose content in D. melanogaster. The addition of activated yeast shortened the developmental time of P.  pseudoannulata at the second instar stage but had no effect on other instars. Different yeast treat-ments in the medium had no effect on the body length or body weight of P.  pseudoannulata. Adding yeast to D. melanogaster culture medium can increase the total fat content in P.  pseudoannulata, but it has no effect on glucose and total protein in P.  pseudoannulata. Our study shows the importance of yeast to the growth and development of fruit flies. Full article

Diacylglycerol-O-acyltransferase 1 (DGAT1, EC 2.3.1.20) is a pivotal enzyme in plant triacylglycerol (TAG) biosynthesis. Previous work identified conserved di-arginine (R) motifs (R-R, R-X-R, and R-X-X-R) in its N-terminal cytoplasmic acyl-CoA binding domain. To elucidate their functional significance, we engineered R-rich sequences in the N-termini of Tropaeolum majus and Zea mays DGAT1s. Comparative analysis with their respective non-mutant constructs showed that deleting or substituting R with glycine in the N-terminal region of DGAT1 markedly reduced lipid accumulation in both Camelina sativa seeds and Saccharomyces cerevisiae cells. Immunofluorescence imaging revealed co-localization of non-mutant and R-substituted DGAT1 with lipid droplets (LDs). However, disruption of an N-terminal di-R motif destabilizes DGAT1, alters LD organization, and impairs recombinant oleosin retention on LDs. Further evidence suggests that the di-R motif mediates DGAT1 retrieval from LDs to the endoplasmic reticulum (ER), implicating its role in dynamic LD–ER protein trafficking. These findings establish the conserved di-R motifs as important regulators of DGAT1 function and LD dynamics, offering insights for the engineering of oil content in diverse biological systems. Full article

The innate immune response is an important defense against invading pathogens. Stimulator of interferon gene (STING) plays an important role in the cyclic GMP-AMP synthase (cGAS)-mediated activation of type I IFN responses. However, some viruses have evolved the ability to inhibit the function of STING and evade the host antiviral defenses. Understanding both the mechanism of action and the viruses targets of STING effector is important because of their importance to evade the host antiviral defenses. In this study, the STING (IpSTING) of Ictalurus punctatus was first identified and characterized. Subsequently, the yeast two-hybrid system (Y2HS) was used to screen for proteins from channel catfish virus (CCV, Ictalurid herpesvirus 1) that interact with IpSTING. The ORFs of the CCV were cloned into the pGBKT7 vector and expressed in the AH109 yeast strain. The bait protein expression was validated by autoactivation, and toxicity investigation compared with control (AH109 yeast strain transformed with empty pGBKT7 and pGADT7 vector). Two positive candidate proteins, ORF41 and ORF65, were identified through Y2HS screening as interacting with IpSTING. Their interactions were further validated using co-immunoprecipitation (Co-IP). This represented the first identification of interactions between IpSTING and the CCV proteins ORF41 and ORF65. The data advanced our understanding of the functions of ORF41 and ORF65 and suggested that they might contribute to the evasion of host antiviral defenses. However, the interaction mechanism between IpSTING, and CCV proteins ORF41 and ORF65 still needs to be further explored. Full article

This study aimed to evaluate the production of xylose reductase (XR), an enzyme responsible for converting xylose into xylitol, by Candida tropicalis ATCC 750 using hemicellulosic hydrolysate from cashew apple bagasse (CABHM) as a low-cost carbon source. The effects of temperature, aeration, and fluid dynamics on XR biosynthesis were also investigated. The highest XR production (1.53 U mL⁻¹) was achieved at 30 °C, with 8.3 g·L⁻¹ of xylitol produced by the yeast under microaerobic conditions, demonstrating that aeration and fluid dynamics are important factors in this process. Cellular metabolism and enzyme production decreased at temperatures above 35 °C. The maximum enzymatic activity was observed at pH 7.0 and 50 °C. XR is a heterodimeric protein with a molecular mass of approximately 30 kDa. These results indicate that CABHM is a promising substrate for XR production by C. tropicalis, contributing to the development of enzymatic bioprocesses for xylitol production from lignocellulosic biomass. This study also demonstrates the potential of agro-industrial residues as sustainable feedstocks in biorefineries, aligning with the principles of a circular bioeconomy. Full article

The objective of this study was to evaluate the effects of feeding a combination of chicory–plantain silage and supplementing Se yeast on the response of early-lactating ewes to induce subclinical mastitis. Polypay ewes (n = 32) were fed either chicory–plantain silage or grass silage and supplemented with 3.6 mg Se yeast/ewe/day for approximately 2 months prior to the infusion of S. uberis into both mammary glands (i.e., intramammary infection or IMI). The ewes had a typical subclinical mastitis response with an 8-fold increase in milk somatic cell count within 24 h post-IMI, a decrease in milk yield, and changes in all milk components measured. The ewes experienced a mild systemic inflammation post-IMI as determined by an increase in rectal temperature and decrease in feed and water intake and, in blood, by an increase in the concentration of ceruloplasmin, haptoglobin, and myeloperoxidase and a decrease in paraoxonase, Zn, advanced oxidation protein products, and hematocrit with no effect on pro-inflammatory cytokines. No effect of silage type, likely due to a low concentration of secondary compounds, or Se supplementation was detected in response to IMI. In summary, the subclinical mastitis model used was effective in mounting an inflammatory response, although this was mild; however, feeding chicory–plantain silage with a low concentration of secondary compounds and supplementing Se yeast had no significant effect on the response of ewes to mammary infection. Full article

The food industry consumes large amounts of water across various processes, and generates wastewater characterized by parameters like biochemical oxygen demand, chemical oxygen demand, pH, suspended solids, and nutrients. To meet environmental standards and enable reuse or valorization, treatment methods such as physicochemical, biological, and membrane-based processes are applied. This review focuses on the valorization of food industry wastewater in the biotechnological production of high-value products, with an emphasis on starch-rich wastewater, wineries and confectionery industry wastewater, and with a focus on new technologies for reduces environmental burden but also supports circular economy principles. Starch-rich wastewaters, particularly those generated by the potato processing industry, offer considerable potential for biotechnological valorization due to their high content of soluble starch, proteins, organic acids, minerals, and lipids. These effluents can be efficiently converted by various fungi (e.g., Aspergillus, Trichoderma) and yeasts (e.g., Rhodotorula, Candida) into value-added products such as lipids for biodiesel, organic acids, microbial proteins, carotenoids, and biofungicides. Similarly, winery wastewaters, characterized by elevated concentrations of sugars and polyphenols, have been successfully utilized as medium for microbial cultivation and product synthesis. Microorganisms belonging to the genera Aspergillus, Trichoderma, Chlorella, Klebsiella, and Xanthomonas have demonstrated the ability to transform these effluents into biofuels, microbial biomass, biopolymers, and proteins, contributing to sustainable bioprocess development. Additionally, wastewater from the confectionery industry, rich in sugars, proteins, and lipids, serves as a favorable fermentation medium for the production of xanthan gum, bioethanol, biopesticides, and bioplastics (e.g., PHA and PHB). Microorganisms of the genera Xanthomonas, Bacillus, Zymomonas, and Cupriavidus are commonly employed in these processes. Although there are still certain regulatory issues, research gaps, and the need for more detailed economic analysis and kinetics of such production, we can conclude that this type of biotechnological production on waste streams has great potential, contributing to environmental sustainability and advancing the principles of the circular economy. Full article

In the search for alternative protein sources, single cell proteins have gained increasing attention in recent years. Among them, proteins derived from yeast represent a promising but still underexplored option. To enable their application in food product design, their techno-functional properties must be understood. In order to investigate the impact of precipitation pH on their emulsion-stabilizing properties, yeast proteins from Saccharomyces cerevisiae were isolated via precipitation at different pH (pH 3.5 to 5) after cell disruption in the high-pressure homogenizer. Emulsions containing 5 wt% oil and ~1 wt% protein were analyzed for stability based on their droplet size distribution. Proteins precipitated at pH 3.5 stabilized the smallest oil droplets and prevented partitioning of the emulsion, outperforming proteins precipitated at higher pH values. It is hypothesized that precipitation under acidic conditions induces protein denaturation and thereby exposes hydrophobic regions that enhance adsorption at the oil–water interface and the stabilization of the dispersed oil phase. To investigate the stabilization mechanism, the molecular weight of the proteins was determined using SDS-PAGE, their solubility using Bradford assay, and their aggregation behavior using static laser scattering. Proteins precipitated at pH 3.5 possessed larger molecular weights, lower solubility, and a strong tendency to aggregate. Overall, the findings highlight the potential of yeast-derived proteins as bio-surfactants and suggest that pH-controlled precipitation can tailor their functionality in food formulations. Full article

The global population is increasing rapidly and, according to the United Nations (UN), it is expected to reach 9.8 billion by 2050. The demand for food is also increasing with a growing population. Food shortages, land scarcity, resource depletion, and climate change are significant issues raised due to an increasing population. Meat is a vital source of high-quality protein in the human diet, and addressing the sustainability of meat production is essential to ensuring long-term food security. To cover the meat demand of a growing population, meat scientists are working on several meat alternatives. Bacteria, fungi, yeast, and algae have been identified as sources of microbial proteins that are both effective and sustainable, making them suitable for use in the development of meat analogs. Unlike livestock farming, microbial proteins produce less environmental pollution, need less space and water, and contain all the necessary dietary components. This review examines the status and future of microbial proteins in regard to consolidating and stabilizing the global food system. This review explores the production methods, nutritional benefits, environmental impact, regulatory landscape, and consumer perception of microbial protein-based meat analogs. Additionally, this review highlights the importance of microbial proteins by elaborating on the connection between microbial protein-based meat analogs and multiple UN Sustainable Development Goals. Full article

Bacteriophages (phages), the most abundant biological entities on Earth, have long served as both model systems and therapeutic tools. Recent advances in synthetic biology and genetic engineering have revolutionized the capacity to tailor phages with enhanced functionality beyond their natural capabilities. This review outlines the current landscape of synthetic and functional engineering of phages, encompassing both in-vivo and in-vitro strategies. We describe in-vivo approaches such as phage recombineering systems, CRISPR-Cas-assisted editing, and bacterial retron-based methods, as well as synthetic assembly platforms including yeast-based artificial chromosomes, Gibson, Golden Gate, and iPac assemblies. In addition, we explore in-vitro rebooting using TXTL (transcription–translation) systems, which offer a flexible alternative to cell-based rebooting but are less effective for large genomes or structurally complex phages. Special focus is given to the design of customized phages for targeted applications, including host range expansion via receptor-binding protein modifications, delivery of antimicrobial proteins or CRISPR payloads, and the construction of biocontained, non-replicative capsid systems for safe clinical use. Through illustrative examples, we highlight how these technologies enable the transformation of phages into programmable bactericidal agents, precision diagnostic tools, and drug delivery vehicles. Together, these advances establish a powerful foundation for next-generation antimicrobial platforms and synthetic microbiology. Full article