Search Results (1,924)

There is a growing global demand for cost-effective alternatives to high-priced biologic therapies, which has significantly accelerated the development of biosimilars and positioned them as sustainable and affordable treatment options. Biosimilars include therapeutic products such as monoclonal antibodies, soluble receptors, growth factors, and hormones that demonstrate comparable efficacy, safety, and quality to their reference biologics. By providing lower-cost alternatives, biosimilars play a vital role in bridging the affordability gap and expanding patient access to essential, life-saving treatments, particularly in low- and middle-income countries. This review focuses on current Good Manufacturing Practices (cGMPs) in biosimilar development, highlighting critical processes such as cell line engineering, glycosylation optimization, and bioprocess refinement aimed at improving cell culture productivity and product yield while ensuring consistent safety, efficacy, and quality across production batches. Advances in biotechnology, especially in proteomics and advanced analytical characterization, have improved understanding of cellular mechanisms influencing product quality and strengthened comparability with reference biologics. These scientific innovations have enhanced regulatory and clinical confidence, supporting wider acceptance and use amongst patients. In addition, this review examines the evolving global regulatory landscape governing biosimilars and its role in reducing development timelines and costs. Together, scientific innovation, standardized manufacturing practices, and harmonized regulatory frameworks foster competition, accelerate market entry, and ultimately help bridge the gap between innovation and affordability, ensuring equitable and sustainable global access to advanced biologic therapies. Full article

The recovery of extracellular polymeric substances (EPS) from activated sludge (AS) represents a promising strategy to transform wastewater treatment plants (WWTPs) into resource recovery facilities within a circular economy framework. In this study, EPS was extracted from an AS process in a full-scale WWTP, highlighting its catalytic and bioflocculant properties, which represent an innovation in the valorization of this biopolymer. The EPS was subsequently characterized in terms of polysaccharides, proteins, and enzymatic activities (amylase and lipase). The bioflocculation performance of the EPS was evaluated using activated sludge mixed liquor. Results showed that EPS recovery yields using 50 °C and 80 °C were 196.3 ± 38.2 mg EPS/g sludge and 283.5 ± 85.4 mg EPS/g sludge, respectively. Enzymatic assays confirmed amylase activity ranging from 100 to 350 U/g sludge according to the extraction temperature. Lipolytic activity (20 U/g sludge) was comparable to values reported in the literature for EPS from biological sludge. The addition of EPS significantly improved the sludge settling velocity (from 0.86 to 4.48 m/h) and the sludge volume index (from 118.6 to 35.5). However, EPS application also increased the resistance to filtration by 50% and reduced cellular respiration by approximately 40%. Overall, the findings demonstrate that EPS from activated sludge acts as an effective bioflocculant with relevant catalytic properties, highlighting its potential as a high-value biotechnological product while also pointing to operational challenges that require further optimization. Full article

Bacterial extracellular vesicles (BEVs) are nanosized (10–400 nm), membrane-enclosed particles naturally secreted by both Gram-negative and Gram-positive bacteria. Initially characterized as virulence factors in pathogenic species, BEVs are now recognized as multifunctional entities with significant biotechnological potential. Their cargo—comprising proteins, lipids, nucleic acids, and metabolites—enables diverse biological activities, including immune modulation, epithelial barrier protection, stress tolerance, and intercellular communication. Recent studies have highlighted BEVs from biotechnologically relevant bacteria—such as plant growth-promoting rhizobacteria, lactic acid bacteria, bifidobacteria, cyanobacteria, bacilli, and streptomycetes—for their different roles in biological and ecological interactions. These properties underpin emerging applications in health, agriculture, and bioprocessing, including next-generation postbiotics, vaccine platforms, drug and RNA delivery systems, and novel plant biostimulants. However, major challenges persist, particularly low production yields, variability in cargo composition, and scalability. Addressing these limitations requires a deeper understanding of vesiculation mechanisms and the development of process-oriented strategies for BEV recovery and purification. This review synthesizes recent advances in genetic analysis, physiological modulation, physicochemical stimuli, and bioprocess optimization aimed at enhancing BEV production and stabilizing cargo profiles, providing a comprehensive overview of approaches to unlock the full potential of BEVs as versatile biotechnological tools. Full article

This study investigated a phototrophic approach to close nutrient loops by using landfill leachate as a culture medium to produce biomass and polyhydroxybutyrate (PHB) from a thermotolerant strain of Potamosiphon sp. A multi-cycle reuse scheme in which post-culture leachate was partially replenished with fresh leachate and reused in successive cultivation rounds to increase the biomass concentration (g/L) and the intracellular PHB content (% w/w) was tested. Three operational variables were optimized (leachate replenishment percentage, number of reuse cycles, and sanitation method (autoclaving, UV irradiation, or no treatment)) via the Box–Behnken response surface method. Both response variables were modeled with high predictive accuracy (R² = 0.98 for biomass and R² = 1.00 for PHB content). According to the experimental data, leachate replenishment emerged as the key factor influencing nutrient availability—particularly nitrogen and phosphorus—and thus PHB accumulation. The optimized conditions (2.17% v/v fresh leachate, three reuse cycles, and UV sanitation) yielded predicted values of 0.29 g/L biomass and 3.48% w/w PHB. These results demonstrate the feasibility of a controlled multicycle reuse process that integrates effluent treatment and biopolymer synthesis, offering a low-input, circular biotechnological approach for sustainable leachate valorization. Full article

Sabkhas, hypersaline ecosystems along Kuwait’s coastal zone, are extreme environments that harbor diverse halophilic microorganisms with significant biotechnological potential. Despite this, they remain underexplored, particularly in the context of enzymes that can function under high salinity. The aim of this study is to identify bacterial isolates from Kuwait’s sabkhas that produce α-amylase under extreme environmental conditions and to purify and characterize the resulting halotolerant α-amylase. Among the seven α-amylase-producing isolates, Priestia sp. W243, isolated from Mina Abdullah, exhibited the highest enzyme production under optimal growth conditions of pH 9.0, 37 °C, and 7.5% NaCl. A novel halotolerant α-amylase with a remarkably high specific activity (8112.1 U/mg) was purified from this isolate using ultrafiltration, ion-exchange chromatography, and gel-filtration. The purified enzyme, with a molecular weight of 25 kDa, showed optimal activity at 40 °C, pH 8, and 3% NaCl. Notably, the enzyme remained active in the absence of salt and up to 15% NaCl, demonstrating exceptional halotolerance. Metal ion profiling revealed that enzyme activity was significantly enhanced by Co²⁺, whereas Ca²⁺ had a comparatively moderate effect on enzyme activity. When the effects of metal chelators were examined, EDTA, a strong metal chelator, inhibited the enzyme. However, the enzyme remained active when Ca²⁺ was specifically removed using EGTA, suggesting that this α-amylase may be a cobalt-dependent metalloenzyme, which is an unusual characteristic among known α-amylases. Additionally, the enzyme retained its catalytic activity under reducing conditions (e.g., in the presence of DTT and β-mercaptoethanol), indicating structural stability is independent of disulfide bonds. These unique properties distinguish this α-amylase from typical salt- or calcium-dependent counterparts and highlight its potential for industrial applications in high-salt food processing, baking, brewing, and environmental remediation. Full article

Functional foods are designed to provide health benefits beyond basic nutrition; however, the incorporation of bioactive compounds often impacts flavor, stability, and consumer acceptance, making flavor science a critical challenge in product development. This review explores the biochemical and biotechnological mechanisms underlying the formation and modulation of flavor in functional foods. Advances in biotechnology, including microbial fermentation, enzyme engineering, biocatalyst immobilization, and metabolic optimization, have facilitated the sustainable production of natural flavor compounds with improved sensory profiles. Emerging technologies, including nanoencapsulation, ultrasound-assisted extraction, nanotechnology, artificial intelligence-driven flavor design, and 3D food printing, are also discussed for their roles in enhancing the stability, bioavailability, and controlled release of bioactive and flavor compounds. By integrating biotechnology and flavor science, these approaches offer promising strategies for developing clean-label, sensory-optimized functional foods that meet nutritional needs while satisfying consumer expectations, thereby driving innovation toward healthier and more sustainable food systems. Full article

The scorpion family Buthidae, renowned for its neurotoxin-rich venoms, dominates toxinology, while non-buthid venoms remain largely unexplored. Here, we present a comprehensive proteomic and biochemical characterization of the Amazonian chactid scorpion Brotheas amazonicus venom (BamazV), with emphasis on molecular complexity, proteolytic processing, and peptide diversity. Using an integrative venomics approach that combines molecular mass-based fractionation, reversed-phase chromatography, high-resolution mass spectrometry, N-terminal sequencing, and functional and immunological analyses, we reveal an unexpectedly complex venom profile enriched in high-molecular-weight components and extensively processed peptides, with more than 40 venom peptides sequenced by MS/MS and Edman degradation. The data provide evidence for non-canonical proteolytic events, including the generation of peptides from precursor regions not classically associated with mature venom components. In contrast to the venom of Tityus serrulatus, BamazV displays a “hydrolase-rich, neurotoxin-poor” profile, featuring a catalytically active Group III phospholipase A₂ (BamazPLA₂), a highly active hyaluronidase, metalloproteases, low-mass peptides, and potassium channel toxins. Our results suggest a hydrolytic prey-subjugation strategy, and limited cross-reactivity with commercial antivenom highlighted its distinct structural landscape. Overall, this study advances the understanding of venom evolution and proteolytic diversification in underexplored scorpion lineages, positioning B. amazonicus as a valuable model for investigating alternative venom strategies and identifying novel biotechnological scaffolds. Full article

Fumaric acid is one of the most important bio-based chemicals, with applications in the food, feed, polymer, pulp, and pharmaceutical industries. To overcome the limitations of the current petrochemical production process, alternative methods are being developed. Biotechnological production using wild-type fungi like Rhizopus sp. is a promising alternative. In this study, apple pomace was used as a carbohydrate source for fumaric acid production using Rhizopus arrhizus NRRL 1526. Our focus was on the use of free, non-structurally bound carbohydrates present in high amounts in apple pomace originating from direct apple juice processing. Three processes were compared: pressing, extraction, and a combination of both. Two cultivation strategies were applied: pre-culture and separate upstream biomass production. Using the pre-culture approach, a fumaric acid titer of 68.3 g/L was achieved with a yield of 0.53 g/g and a productivity of 0.29 g/(L·h) from synthetic apple pomace juice. Separate biomass production enabled growth-decoupled fumaric acid production, yielding 50.2 g/L and 79.3 g/L with yields of 0.82 g/g and 0.54 g/g and productivities of 0.17 g/(L·h) and 0.27 g/(L·h) from synthetic and real apple pomace juice, respectively. Thus, the efficient use of apple pomace for the fermentative production of fumaric acid is shown. Full article

Members of the Rosaceae family possess substantial economic and ornamental value, making their effective propagation and genetic improvement critical. Plant regeneration represents a foundational technology for efficient breeding, genetic transformation, functional genomics, molecular breeding, germplasm conservation, and large-scale commercial propagation. The regenerative capacity of explants in many Rosaceae taxa remains limited, despite significant progress. This review systematically synthesized conventional and emerging plant regeneration strategies and critically examined the principal biological and technical constraints affecting regenerative efficiency. A comprehensive comparison was first made among the various genera of the Rosaceae family regarding regeneration processes, environmental conditions, PGRs, exogenous additives, basal media, common obstacles and regeneration suggestions. The application of molecular biotechnology approaches in elucidating the mechanisms underlying regeneration and in enhancing regeneration capacity is also evaluated. Finally, this review assesses the future potential of these advanced technologies for improving regeneration systems in Rosaceae plants, providing a comprehensive reference framework for both academic research and industrial applications. Full article

Vinegar is a traditional fermented food of increasing industrial interest due to its nutritional, sensory, and bioactive properties. This study aimed to develop and optimize a controlled biotechnological process for vinegar production from the Algerian date cultivar Degla Beida, an abundant yet underexploited local resource. Indigenous Saccharomyces cerevisiae strains isolated from date fruits and Acetobacter sp. strains isolated from traditional date vinegar were employed as starter cultures in a two-stage submerged fermentation process, comprising alcoholic fermentation followed by acetic fermentation. Process optimization was carried out using Response Surface Methodology (RSM) based on a Central Composite Design (CCD), evaluating the effects of initial alcoholic degree (4–10% v/v) and yeast extract supplementation (0.2–0.5 g/L). The statistical models showed excellent fitting and predictive reliability (p < 0.0001; R² = 94.1–99.1%). Under optimal conditions (7% v/v initial alcohol, 0.2 g/L yeast extract, 30 °C, pH 5), the process yielded a maximum acetic acid concentration of 72 g/L after 11 days, with 80% fermentation efficiency and complete ethanol depletion. The optimized vinegar exhibited enhanced bioactive properties, with a total phenolic content of 620 mg GAE/100 mL and a DPPH radical scavenging activity of 78%, significantly higher than those of the unfermented juice. These results demonstrate the suitability of Degla Beida dates for vinegar production and highlight the potential of indigenous microbial resources for the sustainable valorization of local raw materials through controlled fermentation processes. Full article

This study evaluated the effects of bacterial fermentation (Lactiplantibacillus plantarum, CHOOZIT^®, and YO-MIX^®) and enzymatic hydrolysis (Protamex^® and Neutrase^® at 1% w/w and 5% w/w) on the proximate composition and antioxidant activity of bee pollen from the Colombian tropical dry forest. Both treatments significantly modified the nutritional profile, increasing moisture content (48–71% fermented; 50–68% hydrolyzed) while reducing protein and carbohydrate fractions. Fermentation produced strain-dependent antioxidant effects: L. plantarum maximized ABTS scavenging, while YO-MIX^® 1:1 achieved the highest DPPH activity. Enzymatic hydrolysis demonstrated superior and more consistent improvements across all assays: Neutrase^® 1% achieved 8.5-fold ABTS enhancement, while Protamex^® 1% maximized DPPH scavenging (8-fold). All enzymatic treatments increased total phenolic content by 70–84%. Protamex^® 1% emerged as the optimal treatment, achieving the highest DPPH activity (2689 µM Trolox equivalents/g pollen), substantial antioxidant enhancement across all assays, and preserved nutritional stability (201 kcal/100 g). These findings support the use of mild enzymatic hydrolysis for valorizing Colombian tropical bee pollen as a functional food ingredient with enhanced bioavailability. Full article

The selection of an optimal antifoam is critical for efficient fermentation, as industrial agents often have detrimental side effects like growth inhibition, while some can enhance productivity. We studied the efficacy of novel silicone–polyol antifoam emulsions for use in fermentation as defoamers. Except for agent 3L10, all antifoams tested did not show inhibition on six bacterial and one fungal culture. Interestingly, agent 3L10 strongly inhibited Gram-positive bacteria (especially Corynebacterium glutamicum) but not Gram-negative strains. A comprehensive evaluation protocol—combining chemical design, cytotoxicity screening across diverse microorganisms, the determination of minimum effective concentrations (MECs), and validation in model bioreactor fermentations—was established. Through this process, 6T80 was identified as a promising antifoam agent for fermentation. It exhibited a low MEC, high emulsion stability, and no cytotoxicity and did not impair growth or recombinant protein production in Bacillus subtilis or Pseudomonas putida fermentations. This study concludes that agent 6T80 is suitable for further application in processes involving Gram-negative and certain Gram-positive hosts. The developed methodology enables the targeted selection of highly efficient and biocompatible antifoams for specific biotechnological processes. Full article

In this study, we examined the enhancement of erythritol production by the Yarrowia divulgata strain 1485. Although erythritol fermentation has been thoroughly investigated in earlier studies, the influence of inoculum ratio has not been comprehensively addressed. Therefore, this parameter was selected as the focus of the present work. Since industrial-scale erythritol production is typically carried out using more efficient fungal strains, further improvements in economic viability are primarily expected through integration with other biotechnological processes, allowing the simultaneous generation of multiple valuable products. To this end, the erythritol fermentation was coupled with microbial pigment production, and the potential recovery of additional compounds—such as biodetergents and cosmetic ingredients—were also explored. Based on the results, the fermentation with a 15% inoculation rate appears to be the most effective, producing 67.9 ± 6.0 g/L of erythritol, and 61.81 ± 0.02 mg/L of pigment was successfully extracted at the end of the pigment fermentation. The cells seem capable of increasing the skin’s moisturizing effect according to our preliminary tests when glass bead cell disruption is used, and the emulsifier has also proven to be effective, maintaining an emulsification index (EI) above 50% even after 24 h. When performing a kinetic model, we found that the measured data matched the model predictions and confirmed optimal inoculation size (15%), providing a solid basis for subsequent techno-economic analysis. The integration of the two basic fermentations (erythritol and pigment) is therefore considered successful, and the Yarrowia divulgata strain appears to have great biotechnological potential. Full article

Reducing nitrous oxide (N₂O) emissions from biological wastewater treatment is critical for achieving low-carbon environmental goals. In this study, a Chlamydomonas reinhardtii -driven algal–bacterial granular sludge system was successfully established in a photo-sequencing batch reactor to enhance nitrogen removal while suppressing N₂O generation. Compact granules formed within 48 days, exhibiting good settling ability (SVI₅/SVI₃₀ = 1.0), an average diameter of 0.5 mm, and a mixed-liquor suspended solid concentration of 2.1 g/L. Algal enrichment was confirmed by an increase in chlorophyll-a to 6.6 mg/g-VSS and substantial accumulation of protein-rich extracellular polymeric substances, which improved granule stability and mass transfer. The system achieved efficient pollutant removal when treating synthetic municipal wastewater, maintaining a chemical oxygen demand removal efficiency of approximately 90% and total nitrogen removal of up to 69.4%, with effluent NH₄⁺-N consistently below 1.6 mg/L. Notably, the N₂O emission factor decreased from 4.2 to 0.4 g N₂O-N/kg N-removed, which is lower than that of conventional activated sludge processes. These results demonstrate the potential of microalgae-driven granulation as a promising low-carbon biotechnology for sustainable wastewater treatment. Full article

Photobioreactor-based microalgae cultivation offers an integrated approach for nutrient-rich wastewater treatment while producing valuable biomass. One of the main microalgae components is proteins, making them a biotechnological target. In this work, to develop efficient and greener extraction methodologies, aqueous two-phase systems (ATPSs) based on natural deep eutectic solvents (NADESs) were evaluated for one-step protein extraction from microalgae cultivated in swine wastewater. Six ATPSs combining two NADES—betaine:levulinic acid (Bet:2LA) and choline chloride:urea (ChCl:2Urea)—and their individual components (Bet or ChCl) with phosphate salts were compared. Systems {NADES + K₃PO₄ + water} were characterized and reported for the first time. Protein recovery yield (PRY) and selectivity (protein-to-carbohydrate mass ratio, R) were assessed for three extraction times and at room temperature. The ATPS {Bet:2LA + K₃PO₄ + H₂O} achieved a PRY of 16.4% and remarkable selectivity after 30 min (R = 2.17 g·g⁻¹), with proteins concentrated in the NADES-rich phase, and negligible recovery in the salt-rich phase. Although the maximum PRY (18.2% at 120 min) was achieved with the precursor betaine, the ATPS with Bet:2LA at 30 min offered an optimal balance between efficiency and process time. With a water content of up to 50%, these systems underscore the potential of NADES-based ATPSs as sustainable platforms for protein recovery. Full article