Search Results (1,243)

This study aimed to evaluate solid-state fermentation (SSF) as a sustainable approach for the simultaneous detoxification of olive pomace (OP) and the production of industrially relevant enzymes. OP, a semisolid byproduct of olive oil extraction, is rich in lignocellulose and phenolic compounds, which limit its direct reuse due to phytotoxicity. A native strain of Aspergillus sp., isolated from OP, was employed as the biological agent, while grape pomace (GP) was added as a co-substrate to enhance substrate structure. Fermentations were conducted at two scales, Petri dishes (20 g) and a fixed-bed bioreactor (FBR, 2 kg), under controlled conditions (25 °C, 7 days). Key parameters monitored included dry and wet weight loss, pH, color, phenolic content, and enzymatic activity. Significant reductions in color and polyphenol content were achieved, reaching 68% in Petri dishes and 88.1% in the FBR, respectively. In the FBR, simultaneous monitoring of dry and wet weight loss enabled the estimation of fungal biotransformation, revealing a hysteresis phenomenon not previously reported in SSF studies. Enzymes such as xylanase, endopolygalacturonase, cellulase, and tannase exhibited peak activities between 150 and 180 h, with maximum values of 424.6 U·g⁻¹, 153.6 U·g⁻¹, 67.43 U·g⁻¹, and 6.72 U·g⁻¹, respectively. The experimental data for weight loss, enzyme production, and phenolic reduction were accurately described by logistic and first-order models. These findings demonstrate the high metabolic efficiency of the fungal isolate under SSF conditions and support the feasibility of scaling up this process. The proposed strategy offers a low-cost and sustainable solution for OP valorization, aligning with circular economy principles by transforming agro-industrial residues into valuable bioproducts. Full article

Background: Prebiotics are selectively used by host microorganisms to promote health. Because effective prebiotic doses (1.5–30 g/day) often require inconvenient delivery formats, this study aims to explore whether capsule-compatible doses of galacto-oligosaccharides (GOS) can effectively modulate the gut microbiome. Methods: The impact of Bimuno^® GOS (Reading, UK) at 0.5, 0.75, 1.83, and 3.65 g on the adult gut microbiome was assessed using the ex vivo SIFR^® technology (n = 8), a clinically validated, bioreactor-based technology. Results: The GOS were rapidly fermented and significantly increased beneficial Bifidobacterium species (B. adolescentis, B. bifidum, and B. longum), even at the lowest tested dose. In doing so, GOS strongly promoted SCFA production, particularly acetate (significant from 0.5 g) and butyrate (significant from 0.75 g). Gas production only mildly increased, likely as Bifidobacterium species do not produce gases. Based on the ability of the SIFR^® technology to cultivate strictly anaerobic, hard-to-culture gut microbes, unlike in past in vitro studies, we elucidated that GOS also enriched specific Lachnospiraceae species. Besides Anaerobutyricum hallii, this included Bariatricus comes, Blautia species (B. massiliensis, Blautia_A, B. faecis), Oliverpabstia intestinalis, Mediterraneibacter faecis, and Fusicatenibacter species. Finally, GOS also promoted propionate (significant from 0.75 g), linked to increases in Phocaeicola vulgatus. Conclusions: GOS displayed prebiotic potential at capsule-compatible doses, offering greater flexibility in nutritional product formulation and consumer convenience. Notably, the strong response at the lowest dose suggests effective microbiome modulation at lower levels than previously expected. Full article

The biopharmaceutical industry is undergoing a fundamental transformation from traditional batch manufacturing to continuous manufacturing (CM) for recombinant drugs and biosimilars, driven by regulatory support through the International Council for Harmonization (ICH) Q13 guidance and compelling economic advantages. This comprehensive review examines the technical, economic, and regulatory aspects of implementing continuous manufacturing specifically for recombinant protein production and biosimilar development, synthesizing validated data from peer-reviewed research, regulatory sources, and global implementation case studies. The analysis demonstrates that continuous manufacturing offers substantial benefits, including a reduced equipment footprint of up to 70%, a 3- to 5-fold increase in volumetric productivity, enhanced product quality consistency, and facility cost reductions of 30–50% compared to traditional batch processes. Leading biomanufacturers across North America, Europe, and the Asia–Pacific region are successfully integrating perfusion upstream processes with connected downstream bioprocesses, enabling the fully end-to-end continuous manufacture of biopharmaceuticals with demonstrated commercial viability. The regulatory framework has been comprehensively established through ICH Q13 guidance and region-specific implementations across the FDA, EMA, PMDA, and emerging market authorities. This review provides a critical analysis of advanced technologies, including single-use perfusion bioreactors, continuous chromatography systems, real-time process analytical technology, and Industry 4.0 integration strategies. The economic modeling presents favorable return-on-investment profiles, accompanied by a detailed analysis of global market dynamics, regional implementation patterns, and supply chain integration opportunities. Full article

Mushrooms are eukaryotic organisms with absorptive heterotrophic nutrition, capable of feeding on organic matter rich in cellulose and lignocellulose. Since ancient times, they have been considered allies and, in certain cultures, they were seen as magical beings or food of the gods. Of the great variety of edible mushrooms identified worldwide, less than 2% are traded on the market. Although mushrooms have been valued for their multiple nutritional and healing benefits, some cultures perceive them as toxic and do not accept them in their culinary practices. Despite the existing skepticism, several researchers are promoting the potential of edible mushrooms. There are two main methods of mushroom cultivation: solid-state fermentation and submerged fermentation. The former is the most widely used and simplest, since the fungus grows in its natural environment; in the latter, the fungus grows suspended without developing a fruiting body. In addition, submerged fermentation is easily monitored and scalable. Both systems are important and have their limitations. This article discusses the main methods used to increase the performance of submerged fermentation with emphasis on the modes of operation used, types of bioreactors and application of morphological bioengineering of filamentous fungi, and especially the use of intelligent automatic control technologies and the use of non-invasive monitoring in fermentation systems thanks to the development of machine learning (ML), neural networks, and the use of big data, which will allow more accurate decisions to be made in the fermentation of filamentous fungi in submerged environments with improvements in production yields. Full article

Membrane bioreactors (MBRs) have been utilized for maricultural wastewater treatment, where high-salinity stress results in dramatic membrane fouling in the actual process. A microalgal–bacterial symbiotic system (MBSS) offers advantages for photosynthetic oxygen production, dynamically regulating the structure of extracellular polymeric substances (EPSs) and improving the salinity tolerance of bacteria and algae. This study centered on the mechanisms of membrane fouling mitigation via the microalgal–bacterial interactions in the MBSS, including improving the pollutant removal, optimizing the system parameters, and controlling the gel layer formation. Moreover, the contribution of electrochemistry to decreasing the inhibitory effects of high-salinity stress was investigated in the MBSS. Furthermore, patterns of shifts in microbial communities and the impacts have been explored using metagenomic technology. Finally, this review aims to offer new insights for membrane fouling mitigation in actual maricultural wastewater treatment. Full article

Bacterial nanocellulose (BNC) is a highly pure biopolymer with promising applications in the biomedical, food, and textile industries. However, the high production costs and low yields obtained in static conditions limit its scalability and industrial applications. This study addresses the sustainable production of BNC using a rotary disk bioreactor (RDB) and explores the use of grape pomace extract as an alternative carbon source for BNC production. Parameters such as the BNC production and biomass yield were evaluated using Komagataeibacter xylinus ATCC 53524 under different operational conditions (disk surface, rotation speed, and number of disks). The results showed that cellulose production increased using silicone-coated disks at 7–9 rpm (up to 2.72 g L⁻¹), while higher yields (5.23 g L⁻¹) were achieved when using grape pomace extract as the culture medium in comparison with conventional HS medium. FTIR and TGA characterizations confirmed that BNC obtained with grape pomace extract presents the same thermal and chemical characteristics than BNC produced with HS medium. This work provides insight into the feasibility of upscaling BNC production using a bioprocessing strategy, combining production in the RDB system and the use of an agro-industrial waste as a sustainable and cost-effective alternative. Full article

The scalability and processability of high-performance membranes remain significant challenges in membrane technology. This work focuses on optimizing the pilot-scale production of patterned polysulfone (PSf) ultrafiltration membranes using the spray-modified non-solvent-induced phase separation (s-NIPS) method on a roll-to-roll pilot line. s-NIPS has already been studied extensively at lab-scale to prepare patterned membranes for various applications including membrane bioreactors (MBR), reverse osmosis (RO) and forward osmosis (FO). Although studied at the lab scale, membranes prepared at a larger scale can significantly differ in performance; therefore, phase inversion parameters, including polymer concentration, molecular weight, and additive type (i.e., polyethylene glycol (PEG) or polyvinylpyrolidine (PVP)) and concentration, were systematically varied when casting on a roll-to-roll, 12″ wide pilot line to identify optimal conditions for achieving defect-free, high-performance, patterned PSf membranes. The membranes were characterized for their pure water permeance, BSA rejection, casting solution viscosities, and resulting morphology. s-NIPS patterned membranes exhibit 150–350% increase in water flux as compared to their reference flat membrane, thanks to very high pattern heights up to 825 µm and formation of finger-like macrovoids. This work bridges the gap between lab-scale and pilot-scale membrane preparation, while proposing an upscaled membrane with great potential for use in water treatment. Full article

Unspecific peroxygenases (UPOs) are promising biocatalysts for oxyfunctionalizations in future sustainable economies and can be efficiently immobilized on the cell surface of their heterologous production yeast. This immobilization has versatile uses, ranging from the mL to m³ scale; but the production of the yeast surface displayed UPOs, and their handling has yet to be optimized to advance sustainable industrial processes in light of the UN’s sustainable development goals. Here, we present optimized production protocols for surface-displayed UPOs for shaken and stirred systems in different scales and describe suitable storage conditions and a sterilization method. We utilized one-factor-at-a-time and design of experiments approaches. We were able to streamline published protocols for shaken flask cultivations to achieve a 60% increase in volumetric activity, using reduced amounts of media. We also show at least a doubling of final activity for bioreactor cultivations by utilizing a different medium than the industry standard. Finally, we present a novel, robust protocol for parallelized methanol-induced enzyme production in Komagataella phaffii in a BioLector XT^® reactor. Enzyme activity did not decrease and even increased by our recommended sterilization method and during storage over 87 days. This study aims to advance the yeast surface display immobilization method by providing methods for efficient production, storage and utilization of this promising biocatalyst. Full article

The batch, repeated batch and fed-batch cultivation strategies, in stirred tank bioreactors, were evaluated to maximize biomass production and the cells’ desulfurization activity (CDA) of Rhodococcus qingshengii IGTS8. The batch culture reached 2.62 g DCW/L biomass, with a productivity of 0.03 g DCW·L⁻¹·h⁻¹ and only 26% glycerol consumption. The repeated batch strategy reduced cultivation time during the first cycle, increasing biomass production by 15%, with 30% glycerol consumed and productivity 2.3 times higher than the batch process; however, subsequent cycles showed no further improvement. CDA peaked early in both modes but declined to 12–13 U/mg DCW by the end of the exponential growth phase. Fed-batch cultivation achieved 8.35 g DCW/L with 87% glycerol consumption, resulting in a threefold increase in volumetric productivity and a 1.7-fold higher specific growth rate compared with the batch mode. CDA remained stable during the fed-batch process and was approximately 40% higher compared with the batch and repeated batch processes. The fed-batch culture was used directly in a two-phase bubble column bioreactor for the desulfurization of dibenzothiophene (DBT), 4-methyl-dibenzothiophene (4-MDBT) and their mixture. The complete desulfurization of 1.4 mM DBT was achieved at a rate of 21.6 mmol DBT/kg DCW/h, while 0.9 mM 4-MDBT was fully converted but at a 2.5-fold lower rate. The simultaneous conversion of the DBT/4-MDBT mixture showed reduced efficiencies of 59.6% and 41.2%, respectively. Full article

Devices that meet the needs of agricultural biogas plants represent a significant share of the energy balance of the source. The digester mixer is a crucial component installed in the fermentation chamber. Energy consumption during mixing depends on the regime and intensity, as well as the rheological properties of the carrier liquid, the dry matter content, and the dimensions of the fibers. Bioreactor operators often oversize mixers and extend mixing duration to avoid disruptions in biogas production. This paper analyzed the influence of digester mixer operations on selected electrical power quality parameters. For this purpose, two agricultural biogas plants with a capacity of 40 kW, connected to the low-voltage grid, were studied (one located approximately 120 m from the transformer station and the second 430 m away). As shown by the correlations presented in the article, the connection point of the biogas plant significantly impacted the magnitude of the influence of mixer operations on the analyzed voltage parameters. In the second biogas plant, switching on the mixers (in the absence of generation) caused the grid voltage to drop to the lower value permitted by regulations. (Switching on the mixers caused a change in voltage by about 30 V.) The most disturbances were introduced into the grid when the power generated by the biogas plant was equal to the power consumed by its internal equipment. (THD_I then reached as high as 63.2%, while in other cases, it did not exceed 17%.) Furthermore, the operation of the mixers alone resulted in a reduction of approximately 1 MWh of energy exported to the power grid per month. Full article

Poly-β-hydroxybutyrate (P(3HB)) represents a suitable alternative for plastic replacement, since it consists of intracellularly produced polyesters by different microorganisms including Bacillus thuringiensis (Bt). P(3HB) conserves most of the properties of petroleum-derived plastics; however, some drawbacks are the production costs, processing times, and bioseparation techniques, limiting its extended use. Bt has production advantages over other microorganisms, such as those growing in conventional or non-conventional substrates, with short periods of fermentation, which make it an interesting candidate to develop optimized production processes. In this work, we identified P(3HB) producers from 72 isolates of Bt, from which we selected four potential candidates. These isolates were cultivated under different carbon:nitrogen (C:N) ratios of 3, 7, 30, and 50 in a complex medium named (CM). Here, the best conditions for growth in Bt isolates were C:N 3 and 7 ratios, whereas for P(3HB) production they were C:N 7 and 30. Following this, an experiment in a bioreactor was conducted with isolate 81C with the selected C:N ratio of 30, where the produced P(3HB) achieved a maximum at 10 h. Fourier transform infrared spectroscopy (FTIR)was used to characterize flask and bioreactor cultures. It must be mentioned that although a higher concentration of medium was used, this did not improve P(3HB) accumulation. This research demonstrates that C:N ratios can differentially influence growth and P(3HB) accumulation in Bt isolates, which can serve as a reference to develop P(3HB) production processes using Bt as a microbial production platform. Full article

The transition toward a circular water economy addresses accelerating water scarcity and pollution. A PRISMA-2020 systematic review of 50 peer-reviewed articles (January 2018–April 2024) mapped current technologies and management strategies, seeking patterns, barriers, and critical bottlenecks. Bibliometric analysis revealed the following three dominant patterns: (i) rapid diffusion of membrane bioreactors, constructed wetlands, and advanced oxidation processes; (ii) research geographically concentrated in Asia and the European Union; (iii) industry’s marked preference for by-product valorization. Key barriers—high energy costs, fragmented regulatory frameworks, and low social acceptance—converge as critical constraints during scale-up. The following three practical action lines emerge: (1) adopt progressive tariffs and targeted tax credits that internalize environmental externalities; (2) harmonize water-reuse regulations with comparable circularity metrics; (3) create multi-actor platforms that co-design projects, boosting local legitimacy. These findings provide policymakers and water-sector practitioners with a clear roadmap for accelerating Sustainable Development Goals 6, 9, and 12 through circular, inclusive, low-carbon water systems. Full article

Background: Modern viral vector production needs to consider process intensification for higher yields from smaller production volumes. However, innate antiviral immunity triggered in the producer cell may limit virus replication. While commonly used cell lines (e.g., Vero or E1A-immortalised cells) are already compromised in antiviral pathways, the redundancy of innate signaling complicates host cell optimization by genetic engineering. Small molecules that are hypothesized to target antiviral pathways (Viral Sensitizers, VSEs) added to the culture media offer a versatile alternative to genetic modifications to increase permissiveness and, thus, viral yields across multiple cell lines. Methods: To explore how the yield for a chimeric Zika vaccine candidate (YF-ZIK) could be further be increased in an intensified bioprocess, we used spin tubes or an Ambr15 high-throughput microbioreactor system as scale-down models to optimize the dosing for eight VSEs in three host cell lines (AGE1.CR.pIX, BHK-21, and HEK293-F) based on their tolerability. Results: Addition of VSEs to an already optimized infection process significantly increased infectious titers by up to sevenfold for all three cell lines tested. The development of multi-component VSE formulations using a design of experiments approach allowed further synergistic titer increases in AGE1.CR.pIX cells. Scale-up to 1 L stirred-tank bioreactors and 3D-printed mimics of 200 or 2000 L reactors resulted in up to threefold and eightfold increases, respectively. Conclusions: Addition of single VSEs or combinations thereof allowed a further increase in YF-ZIK titers beyond the yield of an already optimized, highly intensified process. The described approach validates the use of VSEs and can be instructive for optimizing other virus production processes. Full article

Rouxiella badensis DSM 100043^T had been previously proven to produce a novel glucoselipid biosurfactant which has a very low critical micelle concentration (CMC) as well as very good stability against a wide range of pH, temperature, and salinity. In this study, we performed a function-based library screening from a R. badensis DSM 100043^T genome library to identify responsible genes for biosynthesis of this glucoselipid. The identified open reading frames (ORFs) were cloned into several constructs in Escherichia coli for gene permutation analysis and the individual products were analyzed using high-performance thin-layer chromatography (HPTLC). Products of interest from positive expression strains were purified and analyzed by liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) and nuclear magnetic resonance (NMR) for further structure elucidation. Function-based screening of 5400 clones led to the identification of an operon containing three ORFs encoding acetyltransferase GlcA (ORF1), acyltransferase GlcB (ORF2), and phosphatase/HAD GlcC (ORF3). E. coli pCAT2, with all three ORFs, resulted in the production of identical R. badensis DSM 100043^T glucosedilipid with Glu-C_10:0-C_12:1 as the main congener. ORF2-deletion strain E. coli pAFP1 primarily produced glucosemonolipids, with Glu-C_10:0,3OH and Glu-C_12:0 as the major congeners, predominantly esterified at the C-2 position of the glucose moiety. Furthermore, fed-batch bioreactor cultivation of E. coli pCAT2 using glucose as the carbon source yielded a maximum glucosedilipid titer of 2.34 g/L after 25 h of fermentation, which is 55-fold higher than that produced by batch cultivation of R. badensis DSM 100043^T in the previous study. Full article

The production of β-alanine from fatty acid feedstocks presents a promising synthetic strategy due to its high carbon yield. However, the excessive reducing power generated during fatty acid utilization disrupts cellular redox balance, adversely affecting metabolism and limiting the efficiency and final yield of β-alanine production. To address this challenge, we engineered a co-production system in which excess reducing equivalents generated during fatty acid β-oxidation and β-alanine biosynthesis were consumed by growth-coupled lycopene biosynthesis. The resulting dual-pathway strain, SA01, achieved 44.78 g/L β-alanine and 3.07 g/L lycopene in bioreactor fermentation, representing a 21.45% increase in β-alanine production compared to the β-alanine-producing strain WA01, and a 74.43% increase in lycopene production compared to the lycopene-producing strain LA01. Further optimization in strain SA06, involving cofactor engineering to shift redox flow from NADH to NADPH, enhanced the titers to 52.78 g/L β-alanine and 3.61 g/L lycopene. Metabolite analysis confirmed a decrease in intracellular NADH and FADH₂ levels in SA06, indicating restoration of redox balance during the late fermentation phase. Additional improvements in the fermentation process, including gradual carbon source switching, optimization of the induction strategy, and fine-tuning of conditions during both growth and bioconversion phases, resulted in further increases in product titers, reaching 72 g/L β-alanine and 6.15 g/L lycopene. This study offers valuable insights into the development of microbial co-production systems, highlighting the critical role of dynamic cofactor and redox balance management, as well as process optimization, in improving production efficiency. Full article