Search Results (2,871)

Residual microalgal biomass generated during wastewater phytoremediation represents an underexploited resource for developing sustainable bioprocesses. This study investigated the biotechnological valorization of phytoremediation-derived Chlorella biomass through an integrated process combining α-amylase-assisted saccharification and fermentation with the non-conventional yeast Wickerhamomyces sp. UFFS-CE-3.1.2 in a stirred-tank bioreactor. Following physical pretreatment to enhance intracellular compound accessibility, fermentation was conducted for 72 h under anaerobic conditions, and temporal changes in enzymatic activities and fermentation-associated compounds were monitored by spectrophotometric assays and high-performance liquid chromatography (HPLC), respectively. The integrated process exhibited distinct temporal profiles of hydrolytic and antioxidant enzyme activities, with maximum activities of 1275.23 U/mL for catalase, 1014.59 U/mL for ascorbate peroxidase, 1291.67 U/mL for protease, and 228.75 U/mL for lipase. Amylase activity remained detectable throughout the 72 h process. Total sugars decreased from 8.88 g/L at 0 h to 0.03 g/L at 72 h. In comparison, glycerol peaked at 5.62 g/L at 18 h, and ethanol remained at approximately 1.0 g/L between 18 and 48 h. Because several enzymatic activities were already detected before yeast inoculation, the observed profiles cannot be attributed exclusively to Wickerhamomyces sp. and should instead be interpreted as characteristics of the integrated bioprocess. Overall, the results demonstrate that residual Chlorella biomass generated during wastewater phytoremediation can serve as a renewable feedstock for further biotechnological processing, supporting an extended valorization pathway within a circular bioprocessing framework. Full article

Background: Mammalian arachidonic acid lipoxygenases (ALOXs) are non-heme iron-containing enzymes that oxygenate polyunsaturated fatty acids (PUFAs) with at least two isolated double bonds to hydroperoxy derivatives. The patho-physiological roles of these enzymes in inflammatory, hyperproliferative, and neurological diseases have made them promising targets for pharmacological interventions. Unfortunately, the expression levels of ALOX isoforms in mammalian cells are very low, which makes functional characterization of native enzymes and the development of isoform-specific inhibitors challenging. Methods: Here, we developed a unifying experimental protocol for fed-batch fermentation of mammalian ALOX isoforms in a bioreactor, followed by purification of the recombinant proteins and their functional characterization. Results: Our methodological protocol allowed the preparation of mg amounts of catalytically active human ALOX15, mouse Alox15 and human ALOX15B. The purified proteins are suitable for high-throughput inhibitor screening assays but can also be used as antigens for the preparation of isoform-specific antibodies and for direct structural analyses. Antibodies cross-reacting with human ALOX15 and ALOX15B have been detected in the blood of patients suffering from colorectal cancer. Conclusions: Our functional ALOX data stresses the catalytic differences between mouse and human ALOX15 orthologs, and these catalytic peculiarities need to be considered when the results of mechanistic studies obtained in mouse models of human diseases are transferred to the human situation. Full article

Upgrading biogas to pipeline-quality methane requires the efficient removal of CO₂, yet conventional physicochemical routes remain energy-intensive. Coupling a CO₂-selective membrane with microalgal photosynthetic fixation offers a green alternative, but is constrained by the low CO₂/CH₄ selectivity of common membranes and the poor adhesion of microalgae to hydrophobic membrane surfaces. Here, a dual-functional composite membrane was developed that simultaneously provides CO₂/CH₄ sieving and a biocompatible interface for microalgal attachment, and was integrated into a microalgae membrane bioreactor (MMBR). A cellulose acetate mixed-matrix membrane incorporating polyethyleneimine-grafted ZIF-8 (CA/PZIF-8(15)) achieved a mixed-gas CO₂ permeability of 122.3 Barrer and a CO₂/CH₄ selectivity of 41.17. An ionic-liquid-modified chitosan (CS/IL) coating, first optimized on a commercial flat-sheet polyethersulfone (PES) membrane used as a model surface for the adhesion study, reversed the surface charge from −30.8 to +3.75 mV, lowered the water contact angle to 51.2°, and increased the day-7 adhesion of Scenedesmus obliquus by ~108%. Transferring the coating onto CA/PZIF-8(15) further raised the permeability to 138 Barrer and the selectivity to 57.31, placing the composite above the 2008 Robeson upper bound. In the MMBR, CH₄ purity reached 95.13% after 48 h; a mass balance on the recirculating gas volume indicated that essentially all of the CO₂ removed from the gas phase permeated the membrane, of which an estimated 2% was fixed into microalgal biomass while the remainder was retained in the liquid phase. This work offers a membrane-design strategy that bridges gas-separation functionality and microalgal carbon fixation for sustainable biogas upgrading. Full article

This research investigates the potential for valorizing submerged arc welding (SAW) slag as a partial substitute for Portland cement in eco-efficient mortars. The research focuses on microstructural characterization, mechanical performance, and, innovatively, resistance to microbial adhesion by Escherichia coli. The residue was processed by grinding and incorporated at substitution levels of 5%, 10%, and 15% by weight. The mechanical results demonstrate that the 5% substitution (AM5%) showed the best performance, reaching 47.55 MPa at 28 days, an increase of approximately 20% compared to the reference sample. This improvement is attributed to the pozzolanic effect with the filling and refinement of the pore structure, in the production of the secondary C-A-S-H phase. In the bioreactor biological assay, the AM5% sample demonstrated a significant reduction in bacterial colonization adhering to the material’s surface, with values of 9.4 × 10¹ CFU/cm². Scanning electron microscopy (SEM) analyses revealed that the denser surface, with lower porosity, hindered the anchoring of E. coli and the formation of biofilm. The study concludes that the use of 5% SAW slag not only improves the structural properties of mortars but also increases their durability in environments prone to biological contamination, such as sanitation systems, promoting a sustainable alternative for the management of industrial waste in civil construction. Full article

Lotus corniculatus is a superior leguminous forage with multiple values including forage, ecological, ornamental and medicinal uses. It is also an ideal material for plant bioreactors. As a core technical approach, genetic transformation overcomes the constraints of traditional breeding and facilitates the targeted improvement in stress resistance and agronomic traits in this species. This review summarizes the research progress of the Agrobacterium-mediated genetic transformation of L. corniculatus, focusing on key procedures such as explant selection, strain selection, infection and co-cultivation regimes, basal medium composition, phytohormone regulation, as well as bacteria elimination and transformant screening strategies. We further elaborate on the applications of this transformation system in enhancing tolerance to abiotic stresses (salt, drought and heat), regulating quality-related traits, and developing plant-based vaccine bioreactors. Additionally, this paper critically discusses the major bottlenecks and challenges constraining existing genetic transformation systems in L. corniculatus, and evaluates the prospects for establishing high-efficiency and genetically stable transformation platforms. This review aims to provide theoretical foundations and technical references for germplasm innovation, molecular breeding and comprehensive utilization of L. corniculatus. Full article

To ensure large-scale production of plant-derived polyphenols, efficient and scalable in vitro culture systems are needed that can enable high biomass accumulation and secondary metabolite biosynthesis. The present study evaluates the suitability of transformed Salvia bulleyana shoot and hairy root cultures for biomass growth and phenolic compound production in different bioreactor systems, including two temporary immersion systems (PlantForm™ and RITA^®) and a nutrient sprinkle bioreactor (NSB). A pronounced system- and organ-specific response was observed. In transformed shoot cultures, the RITA^® bioreactor promoted high growth and volumetric productivity, although metabolite accumulation was lowered by partial hyperhydricity. In contrast, the PlantForm™ system provided more stable morphogenesis and higher polyphenol content, although less effective growth. In hairy root cultures, the NSB proved to be the most effective system, combining high biomass productivity with greater phenolic compound accumulation; total polyphenol content reached 65.1 mg/g DW, with rosmarinic acid (RA) constituting up to 78% of total phenolics. The highest volumetric productivity was achieved in NSB-grown hairy roots, reaching 837.9 mg/L total polyphenols and 651.3 mg/L RA. Secondary metabolism was further enhanced by elicitation with 100 μm methyl jasmonate applied under optimized bioreactor conditions. In transformed shoot cultures, elicitation increased RA accumulation by 63%, resulting in final productivity of 1164 mg/L RA, and 1250 mg/L total polyphenols. In hairy root cultures, RA accumulation increased by 44%, leading to final productivity of 795 mg/L RA and 933 mg/L total polyphenols. Our findings indicate that the balance between biomass growth and secondary metabolism in S. bulleyana cultures is determined by bioreactor configuration. The nutrient sprinkle bioreactor represents a highly effective platform for phenolic acid production in hairy root cultures, whereas temporary immersion systems can be successfully applied for transformed shoot cultivation. The combination of tailored bioreactor strategies with methyl jasmonate elicitation provides a promising approach for scalable production of high-value phenolic compounds in plant in vitro culture systems. Full article

Chinese Baijiu solid-state fermentation operates as a heterogeneous reaction bed in which heat, moisture, O₂, acidity, ethanol, and mass-transfer conditions vary across space and time. Because aroma-related microorganisms respond differently to these local stresses, abundance data alone cannot identify which populations remain active or contribute to ester formation at a particular layer, stage, or batch. This review therefore focuses on stress-tolerant functional units that sustain acid–alcohol–ester conversion and argues for spatially resolved sampling combined with activity-, enzyme-, and metabolite-based evidence. As a conceptual verification platform, rather than a reported experimental apparatus or a replacement for traditional pits, a gradient solid-state bioreactor could reproduce selected temperature, O₂, moisture, and interface gradients under controlled conditions. Coupling multipoint temperature, CO₂/O₂, moisture, and acidification-related signals with spectral fingerprints and volatile measurements, together with offline biological assays, could help define functional windows and improve the mechanistic reproducibility of Baijiu fermentation. Full article

Heterotrophic denitrification is increasingly used in land-based recirculating aquaculture systems (RASs) to remove nitrate and minimise new-water demand, but the carbon sources traditionally used—methanol and acetic acid—raise safety, corrosion, and cost concerns. This study evaluated glycerol as an alternative external carbon source in a commercial land-based Atlantic salmon RAS in Norway. Trials in two identical post-smolt systems with integrated denitrification bioreactors compared glycerol (80%) against acetic acid for denitrification performance, nitrite accumulation, pH stability, carbon-source consumption, cost, and hazard profile. Glycerol achieved similar or higher apparent total dissolved nitrogen (TDN) removal than acetic acid and, once the biofilm had acclimated, was completely consumed. It maintained a stable outlet pH (≥6.5), avoiding the enzyme inhibition seen when acetic acid depressed pH, and proved about 1.5 times more cost-effective (≈NOK 66 vs. 97 per kg TDN removed). A standardised hazard assessment ranked glycerol as the safest option for flammability, toxicity, and corrosiveness. TDN removal correlated strongly with dissolved nitrogen gas saturation at the bioreactor outlet (r² = 0.79–0.82), indicating that total gas pressure monitoring is a promising, low-cost proxy for performance. Nitrite accumulation per unit of nitrogen removed was, however, about three times higher under glycerol (≈1 mg NO₂-N per mg TDN) than under acetic acid (≈0.3 mg NO₂-N per mg TDN), and the nine-day glycerol phase was too short to establish whether this resolves or persists as the biofilm matures. Subject to routine nitrite monitoring in the fish tanks, glycerol is a cost-effective, low-hazard, and pH-stable alternative to acetic acid for nitrate removal in salmon RASs. Full article

The integration of coupled dark fermentation (DF) and photofermentation (PF) systems represents a promising approach for concurrent H₂ production and organic load reduction in carbohydrate–rich wastewater. However, their behavior under non–optimized, continuous industrial operation remains inadequately characterized, hindering practical implementation. This case study evaluated an integrated DF–PF system treating a synthetic sugar mixture mimicking acidic fruit and dairy processing wastewater. The system presented herein (5-L reactors) serves as an initial prototype to facilitate scaling to both pilot (40-L reactors) and, ultimately, industrial (600-L) scales using real effluents within the framework of a research project. The bioreactors were operated continuously with a 10 h hydraulic retention time and an organic loading rate of 2.5 g COD L⁻¹ d⁻¹. The pH and temperature were monitored but intentionally left uncontrolled. The DF stage facilitated by hydrogen–producing bacteria achieved H₂ concentrations of up to 57% (v/v) and a maximum production rate of 177 mL H₂ L⁻¹ d⁻¹; however, it demonstrated notable process instability due to the absence of controls. In contrast, the PF stage exhibited negligible H₂ production (1.8% v/v) attributable to the displacement of Rhodopseudomonas species by microbial competition, thereby functioning predominantly as a polishing step. Overall, the coupled system achieved an average COD removal of 34%, highlighting the functional differentiation between stages and identifying microbial competition as the primary constraint under suboptimal conditions. Full article

Chronic kidney disease (CKD), one of the major global public health burdens, continues to exhibit a rising prevalence worldwide. The growing population of patients with end-stage renal disease (ESRD) has led to an increasing demand for renal replacement therapy (RRT). Although dialysis effectively prolongs survival, it fails to fully replicate kidney function. In addition, the persistent shortage of donor kidneys results in prolonged waiting periods for kidney transplantation. Emerging renal replacement strategies, such as kidney organoids, have demonstrated considerable potential. However, multiple technical limitations continue to hinder their near-term clinical translation. Bioartificial kidneys (BAKs), which integrate engineering and biological technologies, generally consist of artificial filtration membranes and living-cell bioreactors designed to mimic native kidney function. Advances in nanotechnology, biomaterials, and tissue engineering have accelerated the development of implantable bioartificial kidneys (iBAKs), making them an important research direction in renal replacement therapy. These innovations have improved membrane performance, biocompatibility, and cellular integration; however, substantial challenges remain regarding long-term stability, immune compatibility, and clinical validation before translation into human applications. Specifically, limited cell sources and uncertain long-term biocompatibility remain major barriers to iBAK development. In the future, biosensors and artificial intelligence (AI) technologies may be incorporated into bioartificial kidneys to enable personalized precision therapy. This review focuses on the developmental and major challenges of bioartificial kidneys, with detailed discussion of recent progress in implantable artificial kidneys. Full article

Transforming traditional membrane bioreactors (MBRs) into forward osmosis membrane bioreactors (OMBRs) is a highly challenging yet promising technological upgrade. Although both combine biological treatment and membrane separation, their core driving forces and operating mechanisms are completely different (an MBR is pressure-driven, while an OMBR is osmosis-driven). In this paper, a novel OMBR with an integrated fixed biofilm (BF-OMBR) was tested for the treatment of synthetic rural wastewater using fertilizer potassium chloride (KCl) as the draw solution (DS) and a commercial aquaporin Inside^TM forward osmosis (FO) membrane. A bench-scale investigation was conducted to compare the BF-OMBR with traditional OMBRs and MBRs. The experimental data suggested that the reactor with aquaporin membranes contributed to a higher water flux than traditional TFC membranes, while immobilized biofilms improved the total nitrogen removal rate compared to normal OMBRs. The integration of these two technologies in the BF-OMBR system appears to leverage these individual benefits. Its TOC and ammonia nitrogen removal efficiencies were also better than those of the other two bioreactors. Meanwhile, the BF-OMBR successfully controlled the salinity build-up to a level not exceeding 2.5 mS/cm over 90 days of operation. This novel osmotic bioreactor may represent a possible alternative approach to overcoming the challenges of low-temperature and low-C/N-ratio rural sewage treatment. Full article

The industrial use of non-Saccharomyces yeasts offers a robust bioprocess strategy for modulating volatile biochemical profiles and reducing climate-driven ethanol production in fermented beverages. This study explores the metabolic diversity of indigenous non-Saccharomyces, with a particular focus on Starmerella bacillaris MI151, selected for sequential inoculation with Saccharomyces cerevisiae MI118. Pilot-scale bioreactor fermentations (25 L) of two Romanian matrix-specific aromatic grape cultivars, Busuioacă de Bohotin and Tămâioasă Românească, exhibited clear strain-dependent carbon flux changes. The sequential culture actively triggered the glycerol-pyruvic pathway, which consistently reduced the final ethanol concentration to up to 0.94% (v/v) and maintained a balanced profile of volatile compounds. GC-MS-based metabolomic profiling revealed substantial changes in the esterification kinetics and glycosidic precursor cleavage. The sequential fermentation resulted in a synergistic increase in n-hexyl acetate (up to 2310.01 µg/L) and bypassing of the standard enzymatic repression, freeing highly volatile monoterpenes (β-citronellol and β-geraniol) in the analyzed matrices. This targeted microbial system successfully modulated the organic acid–phenolic balance, neutralizing harsh structural finishes, as confirmed by quantitative sensory mapping. Finally, these specific local yeast strains exhibit strong bioprocess scalability, offering a predictable, non-engineered strategy to lower the ethanol yield while driving targeted flavor enhancement in climate-vulnerable viticultural regions. Full article

The temporomandibular joint (TMJ) is a highly specialized synovial joint responsible for essential functions such as mastication, speech, and swallowing. Owing to its unique anatomical organization, complex biomechanics, and heterogeneous tissue composition, regeneration of the TMJ remains one of the greatest challenges in craniofacial reconstructive surgery. Conventional treatment modalities, including autologous grafts, alloplastic prostheses, and total joint replacement, are associated with several limitations, including donor-site morbidity, prosthetic wear, limited biological integration, and the inability to restore native tissue architecture. Three-dimensional (3D) bioprinting has emerged as a promising regenerative strategy capable of fabricating patient-specific living constructs that closely mimic the structural and functional characteristics of the native joint. This review summarizes recent advances in TMJ bioprinting, with particular emphasis on the regeneration of the mandibular condylar fibrocartilage, subchondral bone, articular disc, and integrated osteochondral constructs. The literature search covered publications from January 2010 through March 2026, and it was conducted using major scientific databases, including PubMed/MEDLINE, Scopus, Web of Science, and Google Scholar. Current progress in cellular sources, including mesenchymal stem cells and induced pluripotent stem cells, biomaterials and bioinks, growth factor delivery, and multimaterial bioprinting technologies is discussed. Particular attention is given to the challenges associated with reproducing the complex osteochondral interface, achieving adequate vascularization, ensuring long-term mechanical stability, and directing tissue-specific cell differentiation. Emerging technologies, including four-dimensional (4D) bioprinting, decellularized extracellular matrix-based bioinks, artificial intelligence-assisted design, patient-specific computational modeling, and bioreactor-mediated tissue maturation, are highlighted as promising approaches to improve construct functionality and clinical translation. Although the clinical application of TMJ bioprinting remains in its early stages, rapid advances in regenerative medicine and biofabrication technologies indicate that personalized bioengineered joint reconstruction may become a viable therapeutic option for the treatment of severe temporomandibular joint disorders in the future. Full article

The efficient extraction and purification of recombinant β-casein from Escherichia coli is a crucial step in bioprocess engineering. This study evaluates an adapted high-pressure homogenization (HPH) workflow for recombinant β-casein recovery and compares it with osmotic shock (OS) and standard HPH based on specific protein yield, estimated relative purity, and processing time. In contrast to standard HPH, which requires separate inclusion body recovery, washing, and subsequent denaturant-mediated solubilization, the adapted-HPH workflow integrates mechanical cell disruption with inclusion body recovery during homogenization. Standard HPH was evaluated in two downstream-processing runs, whereas adapted HPH and OS were each evaluated in three downstream-processing runs using separate aliquots from the same bioreactor biomass batch. Adapted HPH demonstrated higher specific yields (4.08 mg_casein/g_CDW) and required 40% less processing time compared to the OS and standard HPH methods. The specific yield obtained with adapted HPH was significantly higher than that obtained with standard HPH (p = 0.026), whereas the difference between adapted HPH and OS was not statistically significant (p = 0.086). Despite the inclusion body washing steps used for OS and standard HPH, adapted HPH achieved a slightly higher average purity than OS (63.46 ± 3.49% versus 59.38 ± 2.79%) and a purity comparable to standard HPH (68.10 ± 6.87%); however, these differences were not statistically significant. Overall, the comparative evaluation of these methods indicates that adapted HPH may provide a simplified alternative for recombinant β-casein recovery, although further validation with a larger number of independent experiments is required. Full article

Extracellular vesicles (EVs) from ovarian cancer cells are valuable sources for candidate biomarker studies, but conventional static flask culture yields limited material and is difficult to scale reproducibly. We evaluated a serum-free CELLine AD 1000 bioreactor workflow for producing EVs from four ovarian cancer-related (OC-related) cell lines (OVCAR4, CaOV3, PA1, SW626) and human dermal fibroblasts (HDFa) as a non-cancer control. Cells were adapted to CDM-HD serum-free medium and maintained for eight weeks with twice-weekly conditioned-medium collection. EVs were isolated by differential ultracentrifugation followed by size-exclusion chromatography and characterized by nanoparticle tracking analysis, imaging flow cytometry, Western blotting, and transmission and scanning electron microscopy. Across longitudinal harvests, OC-related cultures generally produced higher EV particle concentrations and A280-based bulk protein estimates than HDFa, while individual cell lines showed distinct production profiles and membrane-associated growth patterns. A parallel OVCAR4 T-175 flask, maintained in its original serum-containing medium, provided a contextual reference indicating higher per-collection EV particle recovery with the bioreactor, although this was not a matched culture-format comparison. EV-enriched preparations contained vesicle-like particles, with modal diameters of approximately 96–128 nm. Using imaging flow cytometry, the CD9 signal was higher in OC-related EVs and CD63 was most prominent in HDFa; CD9 and CD63 were also detected in OC-related EV lysates by Western blotting. Because one bioreactor was operated per cell line, these findings should be interpreted as preliminary and descriptive rather than statistically comparative. Overall, this study provides a practical serum-free CELLine AD 1000 workflow for generating characterized OC-related EV material for downstream analytical studies. Full article