Search Results (2,647)

Chinese hamster ovary (CHO) cells are the most common protein production platform for glycosylated biopharmaceuticals due to their relatively efficient secretion systems, post-translational modification (PTM) machinery, and quality control mechanisms. However, high productivity and titer demands can overburden these processes. In particular, the endoplasmic reticulum (ER) can become overwhelmed with misfolded proteins, triggering the unfolded protein response (UPR) as evidence of ER stress. The UPR increases the expression of multiple genes/proteins, which are beneficial to protein folding and secretion. However, if the stressed ER cannot return to a state of homeostasis, a prolonged UPR results in apoptosis. Because ER stress poses a substantial bottleneck for secreting protein therapeutics, CHO cells are both selected for and engineered to improve high-quality protein production through optimized UPR and ER stress management. This is vital for optimizing industrial CHO cell fermentation. This review begins with an overview of common ER-stress related markers. Next, the optimal UPR profile of high-producing CHO cells is discussed followed by the context-dependency of a UPR profile for any given recombinant CHO cell line. Recent efforts to control and engineer ER stress-related responses in CHO cell lines through the use of various bioprocess operations and activation/inhibition strategies are elucidated. Finally, this review concludes with a discussion on future directions for engineering the CHO cell UPR. Full article

Agriculture and its related industries produce annually a vast amount of byproducts and waste which comprise a large proportion of global waste. Only a small percentage is managed with environmentally acceptable procedures, while a large proportion is either incinerated or discarded into nearby open fields, causing serious environmental burdens. Since these byproducts exhibit a rich nutritional and phytochemical content, they may be considered as raw materials for various industrial applications, initiating the need for the development of sustainable and eco-friendly methods for their valorization. Among the various methods considered, Solid-State Fermentation (SSF) constitutes an intriguing eco-friendly bioprocess, being suitable for water-insoluble mixtures and providing products with improved stability and depleted catabolic suppression. Thus, there are several literature studies highlighting the aspects and efficacy of SSF for improving the nutritional and phytochemical contents of diverse agro-industrial waste. The review herein aspires to summarize these literature results with a special focus on the enhancement of their antioxidant potency. For this purpose, specific keywords were used for searching multiple scientific databases with an emphasis on the most recent studies and higher impact journals. The presented data establish the usefulness and efficacy of the SSF bioprocess to obtain fermentation products with enhanced antioxidant profiles. Full article

Tailoring culture media and supplementation strategies to the specific requirements of a target product is essential for enhancing microbial production efficiency. This work addresses an unexplored aspect of K. phaffii cultivation: optimizing culture media for metabolite production from xylose, diverging from the conventional focus on recombinant protein expression and the use of glycerol or methanol as primary substrates. Ethylene glycol biosynthesis in an engineered K. phaffii strain was improved by evaluating media and nutrient supplementation. Among the seven evaluated formulations, FM22 and d’Anjou were the most effective, with inositol and thiamine dichloride playing key roles in enhancing production. Salt concentrations in both media were optimized using Central Composite Design (CCD), reducing complexity while increasing yields. Ethylene glycol production increased by 54% in FM22 and 21% in d’Anjou, accompanied by a threefold and 26% reduction in the total salt content, respectively. The vitamin solution was streamlined from seven to two components, each at half the standard concentration. Trace element solutions were reduced to 25% of the original volume without compromising productivity. These findings underscore the dual benefit of culture medium optimization: improved ethylene glycol yields and simplified formulations, establishing a foundation for the development of more efficient and cost-effective bioprocesses using K. phaffii. Full article

The fermentation of Actinobacillus succinogenes in bioelectrochemical systems offers a promising approach to enhance biotechnological succinate production by shifting the redox balance towards succinate and simultaneously enabling CO₂ utilization. Key process parameters include the applied electric potential, electrode material, and reactor design. This study investigates the influence of various carbon fabric electrodes and applied potentials on product distribution during fermentation of A. succinogenes. Building on prior findings that potentials between −600 mV and –800 mV increase succinate production, recent data reveal that more negative potentials, beyond the water electrolysis threshold, trigger electrochemical side reactions, altering product yields. Specifically, succinate decreased from 19.76 ± 0.41 g∙L⁻¹ to 14.1 ± 1.6 g∙L⁻¹, while lactate rose from 0.59 ± 0.12 g∙L⁻¹ to 3.12 ± 0.21 g∙L⁻¹. Contrary to common assumptions, the shift is not primarily driven by oxygen formation. Instead, the results indicate that the intracellular redox potential is affected by both the applied potential and hydrogen evolution, which alters metabolic pathways to maintain redox balance. These findings demonstrate that more negative applied potentials in electro-fermentation processes can impair succinate yields, emphasizing the importance of fine-tuning electrochemical conditions in the system for optimized biotechnological succinate production. Full article

Monascus purpureus is a filamentous fungus renowned for producing bioactive secondary metabolites, including lovastatin and azaphilone pigments. Lovastatin is valued for its cholesterol-lowering properties and cardiovascular benefits, while Monascus pigments exhibit anti-cancer, anti-inflammatory, and antimicrobial activities, underscoring their pharmaceutical and biotechnological relevance. This study evaluated the impact of carbohydrate-derived elicitors—mannan oligosaccharides, oligoguluronate, and oligomannuronate—on the enhancement of pigment and lovastatin production in M. purpureus C322 under submerged fermentation. Elicitors were added at 48 h in shake flasks and 24 h in 2.5 L stirred-tank fermenters. All treatments increased the production of yellow, orange, and red pigments and lovastatin compared to the control, with higher titres upon scale-up. OG led to the highest orange pigment yield (1.2 AU/g CDW in flasks; 1.67 AU/g CDW in fermenters), representing 2.3- and 3.0-fold increases. OM yielded the highest yellow and red pigments (1.24 and 1.35 AU/g CDW in flasks; 1.58 and 1.80 AU/g CDW in fermenters) and the highest lovastatin levels (10.46 and 12.6 mg/g CDW), corresponding to 2.03–3.03-fold improvements. These results highlight the potential of carbohydrate elicitors to stimulate metabolite biosynthesis and facilitate scalable optimisation of fungal fermentation. Full article

Since the mid-19th century, researchers have explored the potential of bio-based polymeric materials for diverse applications, with particular promise in medicine. This review provides a focused and detailed examination of natural and synthetic biopolymers relevant to tissue engineering and biomedical applications. It emphasizes the structural diversity, functional characteristics, and processing strategies of major classes of biopolymers, including polysaccharides (e.g., hyaluronic acid, alginate, chitosan, bacterial cellulose) and proteins (e.g., collagen, silk fibroin, albumin), as well as synthetic biodegradable polymers such as polycaprolactone, polylactic acid, and polyhydroxybutyrate. The central aim of this manuscript is to elucidate how intrinsic properties—such as molecular weight, crystallinity, water retention, and bioactivity—affect the performance of biopolymers in biomedical contexts, particularly in drug delivery, wound healing, and scaffold-based tissue regeneration. This review also highlights recent advancements in polymer functionalization, composite formation, and fabrication techniques (e.g., electrospinning, bioprinting), which have expanded the application potential of these materials. By offering a comparative analysis of structure–property–function relationships across a diverse range of biopolymers, this review provides a comprehensive reference for selecting and engineering materials tailored to specific biomedical challenges. It also identifies key limitations, such as production scalability and mechanical performance, and suggests future directions for developing clinically viable and environmentally sustainable biomaterial platforms. Full article

Cordycepin, a bioactive adenosine analog, holds promise in pharmaceutical and health product development. However, large-scale production remains constrained by the limitations of natural producers, Cordyceps spp. Herein, we report the reconstruction of the first genome-scale metabolic model (GSMM) for a cordycepin-producing strain of recombinant Aspergillus oryzae. The model, iNR1684, incorporated 1684 genes and 1947 reactions with 93% gene-protein-reaction coverage, which was validated by the experimental biomass composition and growth rate. In silico analyses identified key gene amplification targets in the pentose phosphate and one-carbon metabolism pathways, indicating that folate metabolism is crucial for enhancing cordycepin production. Nutrient optimization simulations revealed that chitosan, D-glucosamine, and L-aspartate preferentially supported cordycepin biosynthesis. Additionally, a carbon-to-nitrogen ratio of 11.6:1 was identified and experimentally validated to maximize production, higher than that reported for Cordyceps militaris. These findings correspond to a faster growth rate, enhanced carbon assimilation, and broader substrate utilization by A. oryzae. This study demonstrates the significant role of GSMM in uncovering rational engineering strategies and provides a quantitative framework for precision fermentation, offering scalable and sustainable solutions for industrial cordycepin production. Full article

The analysis of anomalous diffusion characteristics within single-particle tracking data is a key problem in several applied-science domains, including biosignal processing, bioinformatics, and biotechnology. This task becomes particularly challenging in the presence of short trajectories, localization errors, and non-ergodicity, features that are common in real experimental data. To address these limitations, this work proposes an approach that improves the robustness and accuracy of estimating the anomalous diffusion exponent α, even for very short trajectories of up to 10 points. The approach includes an ensemble-based variance estimation of the exponent α, along with a bias correction based on time–ensemble averaged mean squared displacement, which reduces the systematic bias. These components integrate well into neural network architectures and are suitable for analyzing experimental trajectories in biotechnology and bioprocess engineering applications. Full article

A rising demand for low-gluten beer fuels research into enzymatic solutions. This study optimized Aspergillus niger prolyl endopeptidase (AN-PEP) application timing during brewing to reduce gluten while preserving physicochemical quality. Ale-type beers were produced with AN-PEP (2% v/v) added at mashing, boiling, post-boiling, or post-fermentation, plus a control. Three mashing profiles (Mash A, B, C) were also tested. Gluten was quantified by R5 ELISA (LOQ > 270 mg/L). Color, bitterness, ABV, and foam stability were assessed. Statistical analysis involved ANOVA and Tukey’s HSD (p < 0.05). Enzyme activity and thermal inactivation were also evaluated. Initial gluten levels consistently exceeded LOQ. Significant gluten reduction occurred only post-fermentation. Mashing, boiling, and post-boiling additions effectively lowered gluten to below 20 mg/L. Post-fermentation addition resulted in significantly higher residual gluten (136.5 mg/L). Different mashing profiles (A, B, C) with early enzyme addition achieved similar low-gluten levels. AN-PEP showed optimal activity at 60–65 °C, inactivating rapidly at 100 °C. Physicochemical attributes (color, extract, bitterness, ABV) were largely unaffected. However, foam stability was significantly compromised by mashing and post-fermentation additions, while preserved with boiling and post-boiling additions. AN-PEP effectively produces low-gluten beers. Enzyme addition timing is critical: while mashing, boiling, or post-boiling additions reduce gluten to regulatory levels, only the beginning of boiling or post-boiling additions maintain desirable foam stability. These findings offer practical strategies for optimizing low-gluten beer production. Full article

Biodrying and composting are aerobic processes to treat and stabilize organic solid waste, but biodrying involves a shorter process time and does not require the addition of water. The resulting biodried material (BM) is mainly used as an energy source in cement production or in municipal solid waste incineration with energy recovery, but when obtained from agricultural or agroindustrial organic waste, it could also be used as a soil amendment, such as compost (CO). In this study, the phytotoxicity of BM compared to CO, both made from organic wastes (orange peel, mulch and grass), was evaluated on seed germination and growth (for 90 days) of lettuce (Lactuca sativa L.) seedlings on treatments prepared from mixtures of BM and soil, soil (100%) and a mixture of CO and soil. The germination index (GI%) was higher for BM extracts (200 g/L) than for CO extracts (68% vs. 53%, respectively). According to their dry weight, lettuce grew more on the CO mixture (16.5 g) than on the BM (5.4–7.4 g), but both materials far exceeded the soil values (0.15 g). The absence of phytotoxicity suggests that BM acts as a soil amendment, improving soil structure and providing nutrients to the soil. Therefore, biodrying is a quick and low-cost bioprocess to obtain a soil improver. Full article

Background/Objectives: Beak and feather disease virus (BFDV) is the causative agent of psittacine beak and feather disease (PBFD), affecting psittacine birds. There is currently no commercial vaccine or treatment for this disease. This study developed a novel BFDV coat protein mRNA vaccine encapsidated by TMV coat protein to form pseudovirions (PsVs) and tested its immunogenicity alongside BFDV coat protein (CP) subunit and DNA vaccine candidates. Methods: mRNA and BFDV CP subunit vaccine candidates were produced in Nicotiana benthamiana and subsequently purified using PEG precipitation and gradient ultracentrifugation, respectively. The DNA vaccine candidate was produced in E. coli cells harbouring a plasmid with a BFDV1.1mer pseudogenome. Immunogenicity of the vaccine candidates was evaluated in African grey parrot chicks. Results: Successful purification of TMV PsVs harbouring the mRNA vaccine, and of the BFDV-CP subunit vaccine, was confirmed by SDS-PAGE and western blot analysis. TEM analyses confirmed formation of TMV PsVs, while RT-PCR and RT-qPCR cDNA amplification confirmed encapsidation of the mRNA vaccine candidate within TMV particles. Restriction digests verified presence of the BFDV1.1mer genome in the plasmid. Four groups of 5 ten-week-old African grey parrot (Psittacus erithacus) chicks were vaccinated and received two boost vaccinations 2 weeks apart. Blood samples were collected from all four groups on day 14, 28 and 42, and sera were analysed using indirect ELISA, which showed that all vaccine candidates successfully elicited specific anti-BFDV-CP immune responses. The subunit vaccine candidate showed the strongest immune response, indicated by higher binding titres (>6400), followed by the mRNA and DNA vaccine candidates. Conclusions: The candidate vaccines present an important milestone in the search for a protective vaccine against PBFD, and their inexpensive manufacture could considerably aid commercial vaccine development. Full article

In this study, we tested a flexible polyurethane (PU) foam, synthesized from bio-based components, for the removal of petroleum-derived fuels from water samples. The PU was synthesized via the prepolymer method through the reaction of PEG 400 with L-lysine ethyl ester diisocyanate (L-LDI), followed by chain extension with 2,5-bis(hydroxymethyl)furan (BHMF), a renewable platform molecule derived from carbohydrates. Freshwater and seawater samples were artificially contaminated with commercial diesel, gasoline, and kerosene. Batch adsorption experiments revealed that the total sorption capacity (S, g/g) of the PU was slightly higher for diesel in both water types, with values of 67 g/g in freshwater and 70 g/g in seawater. Sorption kinetic analysis indicated that the process follows a pseudo-second-order kinetic model, suggesting strong chemical interactions. Equilibrium data were fitted using Langmuir and Freundlich isotherm models, with the best fit achieved by the Langmuir model, supporting a monolayer adsorption mechanism on homogeneous surfaces. The PU foam can be regenerated up to 50 times by centrifugation, maintaining excellent performance. This study demonstrates a promising application of this sustainable and bio-based polyurethane foam for environmental remediation. Full article

Hybrid modeling techniques are increasingly important for improving predictive accuracy and control in biomanufacturing, particularly in data-limited conditions. This study develops and experimentally validates a hybrid deep learning model predictive control (MPC) framework for recombinant P. pastoris fed-batch fermentations. Bayesian optimization and grid search techniques were employed to identify the best-performing hybrid model architecture: an LSTM layer with 2 hidden units followed by a fully connected layer with 8 nodes and ReLU activation. This design balanced accuracy (NRMSE 4.93%) and computational efficiency (AICc 998). This architecture was adapted to a new, smaller dataset of bacteriophage Qβ coat protein production using transfer learning, yielding strong predictive performance with low validation (3.53%) and test (5.61%) losses. Finally, the hybrid model was integrated into a novel MPC system and experimentally validated, demonstrating robust real-time substrate feed control in a way that allows it to maintain specific target growth rates. The system achieved predictive accuracies of 6.51% for biomass and 14.65% for product estimation, with an average tracking error of 10.64%. In summary, this work establishes a robust, adaptable, and efficient hybrid modeling framework for MPC in P. pastoris bioprocesses. By integrating automated architecture searching, transfer learning, and MPC, the approach offers a practical and generalizable solution for real-time control and supports scalable digital twin deployment in industrial biotechnology. Full article

Alkalihalophilus marmarensis is an obligate alkaliphile with exceptional tolerance to high-pH environments, making it a promising candidate for industrial bioprocesses that require contamination-resistant and extremophilic production platforms. However, its practical deployment is hindered by limited biomass formation under extreme conditions, which constrains overall productivity. This study presents a model-driven investigation of how pH (8.8 and 10.5), culture duration (24 and 48 h), and nitrogen source composition (peptone and meat extract) affect cell dry mass, lactate, and protease synthesis. Using the response surface methodology and multi-objective optimization, we established predictive models (R² up to 0.92) and uncovered key trade-offs in biomass and metabolite yields. Our findings reveal that peptone concentration critically shapes the metabolic output, with low levels inhibiting growth and high levels suppressing protease activity. Maximum cell dry mass (4.5 g/L), lactate (19.3 g/L), and protease activity (43.5 U/mL) were achieved under distinct conditions, highlighting the potential for targeted process tuning. While the model validation confirmed predictions for lactate, deviations in cell dry mass and protease outputs underscore the complexity of growth–product interdependencies under nutrient-limited regimes. This work delivers a foundational framework for developing fermentations with A. marmarensis and advancing its application in sustainable, high-pH industrial bioprocesses. The insights gained here can be further leveraged through synthetic biology and bioprocess engineering to fully exploit the metabolic potential of obligate alkaliphiles like A. marmarensis. Full article

Beer production produces significant amounts of brewers’ spent grain (BSG), a lignocellulosic by-product with important environmental and economic impacts. Despite its high moisture content and rapid microbial breakdown, BSG has a stable, nutrient-rich composition, especially high in protein, fiber, and polyphenolic compounds. While its perishability limits direct use in food systems, BSG is often repurposed as livestock feed. Recent advances in bioprocessing and extraction technologies have expanded their use across different sectors. This review explores the composition of crude BSG and evaluates innovative valorization methods, including recovering bioactive compounds with pharmaceutical and nutraceutical value, and converting them into biofuels such as biogas, biodiesel, and bioethanol. Special focus is given to methods involving enzymatic hydrolysis, fermentation, and chemical extraction to isolate proteins, peptides, amino acids, sugars, and polyphenols. By analyzing emerging applications and industrial scalability challenges, this review highlights BSG’s growing role within circular economy models and its potential to promote sustainable innovations in both the brewing industry and the wider bioeconomy. Full article