Search Results (112)

Starch-based bioplastics are renewable and biodegradable, but their wider use is constrained by moisture sorption, humidity-dependent aging, insufficient tensile performance, and weak water- and oxygen barrier stability. This review critically synthesizes the peer-reviewed literature from 2020 to 2026 on enzymatically engineered starch for film, packaging, and thermoplastic applications using an architecture–property framework that links enzyme specificity, chain-length distribution, crystallinity, processing route, and material response. Controlled α-1,4 hydrolysis mainly improves processability by lowering molecular weight, viscosity, and gelatinization resistance. However, excessive hydrolysis can increase water uptake, solubility, and loss of cohesive strength. Debranching by pullulanase or isoamylase increases amylose-like linear chains and can promote B-type crystallinity or V-type starch–lipid complexes, with reported gains in tensile strength, contact angle, and water vapor barrier when the chain lengths and recrystallization conditions are controlled. Branching enzymes and transglycosylases increase branch density or redistribute glucan chains, suppressing retrogradation and improving flexibility, water retention, and aging resistance, but often with trade-offs in strength, crystallinity, and barrier performance. Lipase- and laccase-catalyzed functionalization expands starch functionality by increasing hydrophobicity, compatibility with hydrophobic phases, antioxidant activity, and active-packaging potential. The evidence indicates that enzymatic modification should not be generalized as uniformly improving starch bioplastics; performance gains are conditional on the starch source, amylose content, enzyme dosage, reaction severity, plasticizer composition, processing method, film conditioning, and storage humidity. Industrial implementation remains limited by enzyme cost and reuse, high-solids mass transfer, reaction time, enzyme stability under heat and shear, and reproducibility across botanical sources. Overall, enzymatic molecular editing is most promising when mechanistic architecture control is coupled with standardized structure–property reporting and scalable processing, such as immobilized-enzyme reactors, high-solids systems, and reactive extrusion. Full article

The textile industry generates dye-containing wastewater characterized by intense coloration, high salinity, elevated oxygen demand, and recalcitrant pollutants. Although conventional treatment technologies can remove color and reduce organic load, decolorization alone does not guarantee detoxification because toxic transformation products may persist or be generated during incomplete degradation. This review emphasizes the synergistic interaction between oxidative and reductive enzymes and microorganisms as a promising strategy for promoting extensive pollutant transformation and potentially enhancing mineralization. Particular attention is given to the complementary roles of laccases (LAC), azoreductases (AZOR), ligninolytic peroxidases, bacteria, fungi, and microbial consortia in sequential degradation pathways that can improve detoxification efficiency. Mechanisms including azo bond cleavage, aromatic amine transformation, and subsequent microbial degradation are discussed to demonstrate how integrated systems can overcome the limitations of enzyme- or microorganism-based treatments alone and may facilitate complete pollutant removal under appropriate conditions. The review also explores biomass-derived biocatalysts produced through solid-state fermentation of agro-industrial residues, demonstrating opportunities to integrate wastewater remediation with biomass valorization and circular bioeconomy principles. Finally, operational challenges, toxicity assessment, reactor design, enzyme immobilization, scale-up, and techno-economic considerations are discussed, and future research priorities are identified to support the development of sustainable and industrially applicable enzymatic–microbial detoxification technologies. Full article

The governing reaction–diffusion model for carbohydrate oxidation catalyzed by an immobilized bienzyme system glucose dehydrogenase and laccase within a spherical porous microreactor is adapted from Baronas et al. and extended here to the non-steady-state regime. The model consists of coupled non-linear partial differential equations based on non-Michaelis–Menten kinetics. The principal novelty of this work lies in the derivation of closed-form semi-analytical expressions for transient and steady-state concentrations of the carbohydrate substrate, oxygen, and product, as well as for the effectiveness factor, using the Laplace Homotopy Perturbation Method (LHPM). The LHPM solutions are validated against MATLAB R2026a numerical simulations (maximum error $<0.009\%$) and demonstrate superior accuracy compared to previously reported Adomian Decomposition Method (ADM) and Taylor Series Method (TSM) solutions. Parametric analysis reveals that the Thiele modulus, saturation parameters, and dimensionless time strongly influence the internal concentration profiles and reactor effectiveness. These analytical results provide rapid, closed-form predictive tools for optimizing catalyst particle size, enzyme loading, and operating conditions in immobilized enzyme microreactor systems. Full article

The growing demand for functional foods reflects greater consumer awareness of diet–health links, with bioactive peptides receiving increasing attention for their health-promoting effects. In this study, bioactive peptides exhibiting antioxidant, dipeptidyl peptidase-IV (DPP-IV) inhibitory, and angiotensin-converting enzyme (ACE) inhibitory activities were produced from a jack bean (Canavalia ensiformis) protein isolate using a continuous proteolysis system with two enzymes. This study encompassed two major phases: isolating protein from jack beans and implementing a continuous enzymatic hydrolysis process. Key variables examined included the enzyme-to-substrate ratio ([E]/[S]), pH level, and residence time (τ). Optimal performance was achieved at [E]/[S] = 5%, pH = 7.5, and τ = 12 h, yielding a permeate with peptide content of 0.6143 mg SE/mL, along with notable antioxidant capacity and ACE inhibition of 0.0454 mg TEAC/mL and 92.18%, respectively. These results confirm that the jack bean protein isolate is a viable substrate for generating multifunctional bioactive peptides. This study provides a foundation for scalable and sustainable production of functional food ingredients from underutilized legumes using continuous bioprocessing technology. Industrial relevance: Integrating a stirred tank reactor with membrane separation provides a promising approach for continuous bioactive peptide production using a free-enzyme system, helping to streamline processing, reduces the demand for enzyme immobilization, and minimizes batch-to-batch variability. This study shows that continuous hydrolysis of jack bean protein isolate in EMR can enhance antioxidant activity and ACE inhibition of the hydrolysates. This approach offers a safer and more efficient route to support the commercialization of jack bean-based functional products. Full article

In this study, the thermodynamic and kinetic characteristics of the esterification reaction catalyzed by sol–gel immobilized enzyme in a batch reactor were systematically investigated. Parameters including catalyst particle diameter, stirring speed, initial molar ratio, catalyst dosage, reaction temperature, and catalyst reusability were studied and optimized. Finally, the esterification rate of methyl oleate reached 90.01%. The standard enthalpy change in the reaction was calculated using the Van’t Hoff equation. A pseudo-homogeneous (PH) model was employed to simulate the kinetic process of the reaction, and the simulation results provide a reference for the scale-up of the reaction process. Full article

Laccases, a class of multicopper oxidases found in diverse biological sources, have emerged as key green biocatalysts with significant potential for eco-friendly pollutant degradation. Their ability to drive electron transfer reactions using oxygen, converting pollutants into less harmful products, positions laccases as promising tools for scalable and sustainable treatment of wastewater, soil, and air pollution. This review explores laccase from a translational perspective, tracing its journey from laboratory discovery to real-world applications. Emphasis is placed on recent advances in production optimization, immobilization strategies, and nanotechnology-enabled enhancements that have improved enzyme stability, reusability, and catalytic efficiency under complex field conditions. Applications are critically discussed for both traditional pollutants such as synthetic dyes, phenolics, and pesticides and emerging contaminants, including endocrine-disrupting chemicals, pharmaceuticals, personal care products, microplastic additives, and PFAS. Special attention is given to hybrid systems integrating laccase with advanced oxidation processes, bioelectrochemical systems, and renewable energy-driven reactors to achieve near-complete pollutant mineralization. Challenges such as cost–benefit limitations, limited substrate range without mediators, and regulatory hurdles are evaluated alongside solutions including protein engineering, mediator-free laccase variants, and continuous-flow bioreactors. By consolidating recent mechanistic insights, this study underscores the translational pathways of laccase, highlighting its potential as a cornerstone of next-generation, scalable, and eco-friendly remediation technologies aligned with circular bioeconomy and low-carbon initiatives. Full article

Enzymes are highly selective and efficient biological catalysts that play a critical role in modern industrial biocatalysis. Their ability to operate under mild conditions and reduce environmental impact makes them ideal alternatives to conventional chemical catalysts. This review provides a comprehensive overview of advances in enzyme-based catalysis, focusing on enzyme classification, engineering strategies, and industrial applications. The six major enzyme classes—hydrolases, oxidoreductases, transferases, lyases, isomerases, and ligases—are discussed in the context of their catalytic roles across sectors such as pharmaceuticals, food processing, textiles, biofuels, and environmental remediation. Recent developments in protein engineering, including directed evolution, rational design, and computational modeling, have significantly enhanced enzyme performance, stability, and substrate specificity. Emerging tools such as machine learning and synthetic biology are accelerating the discovery and optimization of novel enzymes. Progress in enzyme immobilization techniques and reactor design has further improved process scalability, reusability, and operational robustness. Enzyme sourcing has expanded from traditional microbial and plant origins to extremophiles, metagenomic libraries, and recombinant systems. These advances support the integration of enzymes into green chemistry and circular economy frameworks. Despite challenges such as enzyme deactivation and cost barriers, innovative solutions continue to emerge. Enzymes are increasingly enabling cleaner, safer, and more efficient production pathways across industries, supporting the global shift toward sustainable and circular manufacturing. 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

D-allulose is a rare sugar with promising applications in food and health industries, owing to its low caloric value and multiple health benefits. In this study, we systematically investigated a thermostable D-allulose 3-epimerase (TcDAEase) from Thermogemmatispora carboxidivorans for food-compatible continuous production. The enzyme exhibited remarkable thermostability, with over 70% activity retained at 80 °C, and showed broad pH tolerance across the range of 8.0 to 13.0. Notably, TcDAEase exhibited high catalytic activity toward D-allulose and D-fructose even without the addition of metal ions. Moreover, food-grade Mg²⁺ was identified as enhancing enzyme activity by 14.3%, thus ensuring compliance with Generally Recognized as Safe (GRAS) standards for food applications. To improve industrial applicability, the enzyme was immobilized using a chitosan-diatomaceous earth (DE) matrix via three-step adsorption–crosslinking–embedding strategy. The immobilized TcDAEase achieved 48.7% ± 2.4% activity recovery and retained 90.3% ± 1.5% activity over seven reaction cycles. Furthermore, continuous production of D-allulose was realized in a packed-bed reactor, operating stably at 60 °C, pH 8.0 and 5 mM Mg²⁺ for 150 days, producing 756 kg of D-allulose with a conversion yield exceeding 89.7% of the theoretical maximum. Overall, this study provides a feasible strategy for the continuous and efficient production of high-value-added D-allulose in the food industry. Full article

The enzymatic synthesis of statin intermediates offers a sustainable alternative to traditional multistep chemical methods. This study investigates the continuous flow synthesis of statin precursors in a millireactor using 2-deoxy-D-ribose-5-phosphate aldolase (DERA) immobilized on mesoporous silica foam (MCF) and magnetic nanoparticles (MNPs). Two types of flow millireactors, a fixed bed millireactor for MCF and a fluidized bed millireactor for MNP, were designed. Key performance indicators including conversion, selectivity, yield, and productivity were analyzed and compared with the batch reactor results. The MNP-based fluidized bed millisystem demonstrated superior conversion (97.78%) and yield (95.85%) under optimized conditions, outperforming both batch and MCF-based millisystems. This work highlights the importance of optimizing immobilization techniques and reactor configurations to enhance enzyme stability and catalytic efficiency in continuous biocatalytic processes, particularly for pharmaceutical applications. Full article

In this review, we trace the evolution of liquid chromatography from the pioneering work of Tennikova and Svec to the current monolithic polymethacrylate supports for performing liquid chromatography with biological macromolecules and nanoparticles, which offer rapid, high-throughput separations. By using interconnected channels with a tailored channel diameter, monoliths minimize the diffusion limitations typical of particle-based systems. Radial flow designs and optimized channel architectures enable the direct loading of complex biological fluids, reducing the need for sample preparation and optimizing the purification of large biomolecules and nanoparticles such as proteins, nucleic acids, extracellular vesicles, and viruses. Recent work has integrated monoliths into immunoaffinity and enzyme reactor platforms, streamlining analytical workflows and preparative applications in vaccine production and gene therapy. The ongoing advances in monolithic materials, channel geometry, and continuous processing hold promise for even greater efficiency and scalability in future applications. Full article

Background/Objectives: Multi-dimensional liquid chromatography coupled with mass spectrometry (mD-LC-MS) has emerged as a powerful technique for the in-depth characterization of biopharmaceuticals by assessing chromatographically resolved product variants in a streamlined and semi-automated manner. The study aims to demystify and enhance the accessibility to this powerful but inherently complex technique by detailing a robust and user-friendly instrument platform, allowing analysts to switch seamlessly between intact, subunit, and peptide mapping workflows. Methods: Starting from a commercially available Two-Dimensional Liquid Chromatography (2D-LC) system, we introduce specific hardware and software extensions leading to two versatile mD-LC-MS setups, in slightly different configurations. The technique’s efficacy is demonstrated through a case study on a cation exchange chromatography method assessing the charge variants of a bispecific antibody, isolating peak(s) of interest, followed by online sample processing, including reduction and enzymatic digestion, and subsequently mass spectrometry analysis. Results: The accuracy and reproducibility of both mD-LC-MS setups proposed in this study were successfully tested. Despite the complex peak patterns in the first dimension, the systems were equally effective in identifying and quantifying the underlying product species. This case study highlights the routine usability of mD-LC-MS technology for the characterization of (ultra) high-performance liquid chromatography (UHPLC) of therapeutic biomolecule. Conclusions: The demonstrated reliability and accuracy underscore the practicality of mD-LC-MS for routine use in biopharmaceutical analysis. Our detailed description of the mD-LC-MS systems and insights simplify access to this advanced technology for a broader scientific community, regardless of expertise level, and lower the entry barrier for its use in various research and industrial settings. Full article

Electrochemical enzyme biosensors are extensively utilized in clinical analysis and environmental monitoring, yet achieving effective enzyme immobilization while maintaining high activity remains a challenge. In this work, we developed a flow-through enzyme biosensor system using a 3D-printed flow-through electrochemical cell fabricated from commercially available poly (lactic acid). After modification with thiacalixarene-functionalized oligo (lactic acids) (OLAs), the material enabled efficient immobilization of uricase on the inner surface of a replaceable reactor of the cell. Swelling and hydrolytic stability of OLAs in cone, partial cone, and 1,3-alternate conformations were studied, with 1,3-alernate conformation demonstrating superior stability and enzyme immobilization performance. The use of OLAs enhanced immobilization efficiency by over 30% and protected the reactor from swelling, hydrolytic degradation, and enzyme loss. The biosensor was validated for amperometric uric acid determination, with a screen-printed carbon electrode modified with carbon black and Prussian Blue. This modification reduced the cathodic potential for uric acid detection to –0.05 V. The biosensor exhibited a linear detection range of 10 nM to 30 μM with a detection limit of 7 nM, and it performed effectively in artificial urine and synthetic blood plasma. The novel cell design, featuring easy assembly and low-cost replaceable parts, makes this biosensor a promising candidate for routine clinical analysis and other practical applications. Full article

Emerging pollutants such as organophosphate flame retardants (OPFRs) pose a critical threat to environmental and human health, while conventional wastewater treatments often fail to remove them. This study addresses this issue by evaluating the bioremediation potential of white-rot fungi for the removal of two OPFRs: tris(2-chloroethyl) phosphate (TCEP) and tributyl phosphate (TBP). Three fungal species—Ganoderma lucidum, Trametes versicolor, and Phanerochaete velutina—were screened for their degradation capabilities. Among these, G. lucidum and T. versicolor demonstrated removal efficiencies exceeding 99% for TBP, while removal rates for TCEP were significantly lower, with a maximum of 30%. The exploration of the enzyme role showed that cytochrome P450 is involved in the degradation while the extracellular laccase is not involved. Continuous batch experiments were performed using a trickle-bed reactor (TBR) operating under non-sterile conditions, a setting that closely resembles real-world wastewater treatment environments. G. lucidum was immobilized on oak wood chips, and the removal efficiencies were measured to be 85.3% and 54.8% for TBP and TCEP, respectively, over 10 cycles. Microbial community analysis showed that G. lucidum remained the dominant species in the reactor. These findings demonstrate the efficacy of fungal-based trickle-bed bioreactors, offering a sustainable and efficient alternative for addressing environmental pollution caused by highly recalcitrant pollutants. Full article

Immobilized enzyme reactors (IMERs) are critical tools for developing novel oligosaccharides based on the enzymatic catalysis of polysaccharides. In this paper, a novel glucuronan lyase from Peteryoungia rosettiformans was produced, purified, and then immobilized on a CIM^® CDI disk for cleaving glucuronan. The results showed that around 63.6% of glycuronan lyases (800.9 μg) were immobilized on the disk. The V_max values of immobilized glucuronan lyases did not significantly change (56.9 ± 4.7 μM∙min⁻¹), while the K_m values (0.310 ± 0.075 g∙L⁻¹) increased by 2.5 times. It is worth noting that immobilized glucuronan lyases overcame the catalytic inhibition of free enzymes observed under high glucuronan concentrations (0.5–2 g∙L⁻¹). circumscribed central composite design (CCCD) and response surface methodology (RSM) showed that glucuronan concentration, flow rate, and reaction time significantly affected the yield of oligoglucuronans. The degree of polymerization (DP) of degraded glucuronan ranged from DP 2–8 according to the results obtained by high performance anion exchange chromatography coupled with a pulsed amperometric detector (HPAEC-PAD). The IMER retained 50.9% activity after running 2373 column volumes of glucuronan. Finally, this glucuronan lyase reactor was tentatively connected to an immobilized laccase reactor to depolymerize, and gallic acid (GA) was added to glucuronan. Approximately 8.5 mg of GA was added onto 1 g of initial glucuronan, and the GA–oligoglucuronan conjugates showed notable antioxidant activity. Full article