Search Results (598)

Vanillin is the compound of great interest to the industry. It is used to augment and enhance the aroma and taste of food preparations and also as a fragrance compound in perfumes and detergents. Currently, majority of the world’s supply consists of chemically synthesized or lignin-derived vanillin. The application of biocatalysis for sustainable manufacturing of food ingredients, pharmaceutical intermediates, and fine chemicals is the key concept of modern industrial biotechnology. The main goal of this research was to conduct optimization procedures aimed at intensifying the microbial hydrolysis process of the lignin-rich plant raw materials and further bioconversion of the released ferulic acid to vanillin. The tests were performed in the solid-state fermentation system with strains selected during the screening stage on agri-food by-products such as brewer’s spent grain. A specially designed single-use bag bioreactor was used to carry out the process on a preparative scale with the most effective strain. The experiment was designed using the RSM, which allowed for an increase in biosynthesis efficiency from 363 mg/kg to 1413 mg/kg (an increase of 389%). The progress of the process was controlled by the use of chromatographic techniques (HPLC) by quantitative determination of vanillin content in the obtained extracts. Full article

The biochemical conversion of biomass waste and organic slurries into clean methane is a valuable strategy for both reducing environmental pollution and advancing alternative energy sources to support energy security. Anaerobic digestion (AD), a mature renewable technology operated in high-performance bioreactors, continues to attract attention for improvements in energy efficiency, profitability, and long-term sustainability at scale. Recent efforts focus on optimizing biochemical reactions throughout all phases of the anaerobic process while mitigating the production of inhibitory compounds that reduce biodegradation efficiency and, consequently, economic viability. A relatively underexplored but promising strategy involves supplementing fermentation substrates with nanoscale additives to boost biomethane yield. Laboratory-scale studies suggest that nanoparticles (NPs) can enhance process stability, improve biogas yield and quality, and positively influence the value of by-products. This paper presents a comprehensive overview of recent advancements in the application of nanoparticles in catalyzing anaerobic digestion, considering both biochemical and economic perspectives. It evaluates the influence of NPs on bioconversion efficiency at various stages of the process, explores specific metabolic pathways, and addresses challenges associated with recalcitrant biomass. Additionally, currently employed and emerging pre-treatment methods are briefly discussed, highlighting how they affect digestibility and methane production. The study also assesses the potential of various nanocatalysts to enhance anaerobic biodegradation and identifies research gaps that limit the transition from laboratory research to industrial-scale applications. Further investigation is necessary to ensure consistent performance and economic feasibility before widespread adoption can be achieved. Full article

The global increase in municipal and industrial wastewater generation has intensified the need for ecologically resilient and technologically advanced treatment systems. Although traditional biological treatment technologies are effective for organic load reduction, they often fail to remove recalcitrant xenobiotics such as pharmaceuticals, synthetic dyes, endocrine disruptors (EDCs), and microplastics (MPs). Engineered microbial consortia offer a promising and sustainable alternative owing to their metabolic flexibility, ecological resilience, and capacity for syntrophic degradation of complex pollutants. This review critically examines emerging strategies for enhancing microbial bioremediation in wastewater treatment systems (WWTS), focusing on co-digestion, biofilm engineering, targeted bioaugmentation, and incorporation of conductive materials to stimulate direct interspecies electron transfer (DIET). This review highlights how multi-omics platforms, including metagenomics, transcriptomics, and metabolomics, enable high-resolution community profiling and pathway reconstructions. The integration of artificial intelligence (AI) and machine learning (ML) algorithms into bioprocess diagnostics facilitates real-time system optimization, predictive modeling of antibiotic resistance gene (ARG) dynamics, and intelligent bioreactor control. Persistent challenges, such as microbial instability, ARG dissemination, reactor fouling, and the absence of region-specific microbial reference databases, are critically analyzed. This review concludes with a translational pathway for the development of next-generation WWTS that integrate synthetic microbial consortia, AI-mediated biosensors, and modular bioreactors within the One Health and Circular Economy framework. Full article

The primary objective of the present study is to evaluate the effect of conglomerate microorganisms on nitrification in activated sludge. The present study compares this process with activated-sludge technology to explore the variables that influence the complex biochemical processes taking place in bioreactors. The research under consideration involves monitoring the effectiveness of optimizing the wastewater treatment process using kinetic modeling for the nitrification and denitrification processes. The system is designed to simulate various operating scenarios and adjust process parameters in real time. The nitrification rate demonstrates a 99.03% performance, while the denitrification rate ranges from 19.08% to 91.01%. A substantial correlation has been demonstrated between this variable and the temperature of the treated wastewater. This provides the possibility of accurately assessing the ammonium oxidation potential. Furthermore, kinetic equations facilitate the estimation of parameters that are not typically measured, yet are essential for optimizing operational parameters (e.g., dissolved oxygen levels in the aeration tank, sludge dosage, and influent flow rate). This estimation is crucial for enhancing the effectiveness of the process and attaining the desired or anticipated outcomes. This validation underscores the efficacy of the technology, thereby establishing a foundational framework for subsequent research endeavors. These research efforts are directed towards providing decision-makers and stakeholders with actionable insights. The validation underscores the significance of optimized practices in the context of water resource protection. Moreover, it signifies a substantial advancement in the instrumentation of wastewater treatment plants. Full article

Valorization of ricotta cheese exhausted whey (RCEW), a dairy by-product generated in large quantities worldwide, is essential to mitigate its environmental impact and unlock its economic potential. This study explores the use of RCEW as a substrate for polyhydroxyalkanoate (PHA) production by Azohydromonas lata DSM 1123. The substrate was characterized by low protein and fat contents and a relevant lactose concentration (3.81%, w/v). Due to A. lata’s inability to directly metabolize lactose, β-galactosidase supplementation was necessary. Mineral supplementation of pasteurized RCEW significantly improved both microbial biomass and PHA synthesis, achieving up to 25.94% intracellular PHA content, whereas pre-adaptation trials failed to enhance strain performance. Moderate nitrogen limitation in the substrate (C/N ratio 44) favored PHA synthesis (0.55 g/L) and 32.74% intracellular accumulation. Thermal treatments decreased initial microbial contamination, hence a balanced mixture of pasteurized–sterilized (75:25) substrate was used to modulate RCEW protein content without the inclusion of additional technological or chemical processing steps and without lactose loss or dilution. Bioreactor trials using optimized RCEW pre-treatment conditions led to a further increase in biomass (2.36 g/L) and PHA production (0.88 g/L), especially under fed-batch conditions. The extracted polymer was confirmed to be polyhydroxybutyrate (PHB), with high thermal stability and a molecular weight of 5.9 KDa. Full article

This study evaluates the enzymatic hydrolysis of pretreated corn cobs (PCCs) using a blend of commercial enzymes (Cellulase enzyme blend and Viscozyme L), followed by simultaneous saccharification and fermentation (SSF) with Mucor indicus DSM 2185 for ethanol production. A combination of 2% (vol vol⁻¹) Cellulase enzyme blend and 5.18% (vol vol⁻¹) Viscozyme L, corresponding to an enzyme loading of 48.9 FPU/g_PCCs, enabled near-complete hydrolysis of 40 g L⁻¹ PCCs within 6–48 h, achieving 92.66% total carbohydrate conversion into fermentable sugars. In SSF experiments conducted in Erlenmeyer flasks, optimal ethanol production in matrix nutrient medium (MNM) reached 14.95 g L⁻¹, with a conversion coefficient of 0.373 g g⁻¹ at 30 °C over a 48 h period. Scale-up of the bioprocess in a 1.5 L stirred-tank bioreactor at 30 °C resulted in an ethanol concentration of 16.46 g L⁻¹, a total carbohydrate conversion of 86.27%, and a substrate-to-ethanol conversion coefficient of 0.44 g g⁻¹ within 22 h. Minor secondary metabolites, including 0.88 g L⁻¹ xylitol and 0.26 g L⁻¹ glycerol, were also detected. Overall, the results demonstrate the potential of M. indicus in combination with commercial enzyme blends as a scalable strategy for industrial ethanol production. 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

As the core equipment for efficient wastewater treatment, the internal structure of microporous aeration bioreactors directly determines the mass transfer efficiency and treatment performance. Based on Computational Fluid Dynamics (CFD) technology, this study explores the optimization mechanism of a Spherical Grid-Structured on the internal flow field of the reactor through a 3D numerical simulation system, aiming to improve the aeration efficiency and resource utilization. This study used a combination of experimental and numerical simulations to compare and analyze different configurations of the Spherical Grid-Structure. The simulation results show that the optimal equilibrium of the flow field inside the reactor is achieved when the diameter of the grid sphere is 2980 mm: the average flow velocity is increased by 22%, the uniformity of the pressure distribution is improved by 25%, and the peak turbulent kinetic energy is increased by 30%. Based on the Kalman vortex street theory, the periodic vortex induced by the grid structure refines the bubble size to 50–80 microns, improves the oxygen transfer efficiency by 20%, increases the spatial distribution uniformity of bubbles by 35%, and significantly reduces the dead zone volume from 28% to 16.8%, which is a decrease of 40%. This study reveals the quantitative relationship between the structural parameters of the grid and the flow field characteristics through a pure numerical simulation, which provides a theoretical basis and quantifiable optimization scheme for the structural design of the microporous aeration bioreactor, which is of great significance in promoting the development of low-energy and high-efficiency wastewater treatment technology. 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

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

Mandelic acid (MA), as an important chiral aromatic hydroxy acid, is widely used in medicine, the chemical industry, and agriculture. With the continuous growth of market demand, traditional chemical synthesis methods are increasingly inadequate to meet the requirements of green and sustainable development due to issues such as complex processes, poor stereoselectivity, numerous byproducts, and serious environmental pollution. MA synthesis strategies based on biocatalytic technology have become a research hotspot due to their high efficiency, environmental friendliness, and excellent stereoselectivity. Significant progress has been made in enzyme engineering modifications, metabolic pathway design, and process optimization. Importantly, biomechanical research provides a transformative perspective for this field. By analyzing the mechanical response characteristics of microbial cells in bioreactors, biomechanics facilitates the regulation of relevant environmental factors during the fermentation process, thereby improving synthesis efficiency. Molecular dynamics simulations are also employed to uncover stability differences in enzyme–substrate complexes, providing a structural mechanics basis for the rational design of highly catalytically active enzyme variants. These biomechanic-driven approaches lay the foundation for the future development of intelligent, responsive biosynthesis systems. The deep integration of biomechanics and synthetic biology is reshaping the process paradigm of green MA manufacturing. This review will provide a comprehensive summary of the applications of MA and recent advances in its biosynthesis, with a particular focus on the pivotal role of biomechanical characteristics. Full article

Aerated accumulation bioreactors represent a promising alternative for the aerobic bioremediation of solid contaminated substrates. However, achieving homogeneous mixing and effective air distribution remains a key design challenge in solid-phase systems. This study presents the design and construction of a novel pilot-scale aerated bioreactor equipped with an angled-paddle agitation system, specifically developed to improve solid mixing and aeration. To evaluate the geometric configuration, a series of simulations were performed using the Discrete Element Method (DEM), with particle dynamics analyzed through the Lacey Mixing Index ($LMI$). Four paddle angles (0°, 15°, 45°, and 55°) were compared, with the 45° configuration achieving optimal performance, reaching $LMI$ values above 0.95 in less than 15 s and maintaining high homogeneity at a filling volume of 70%. These results confirm that the paddle angle significantly influences mixing efficiency in granular media. While this work focuses on engineering design and DEM-based validation, future studies will include experimental trials to evaluate biodegradation kinetics. The proposed design offers a scalable and adaptable solution for ex situ bioremediation applications. This work reinforces the value of integrating DEM simulations early in the bioreactor development process and opens pathways for further optimization and implementation in real-world environmental remediation scenarios. 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

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