Search Results (40)

Enzymes, as nature’s precision biocatalysts, hold transformative potential across industrial, environmental, and biomedical sectors. However, their instability, solvent sensitivity, and limited reusability in their free form necessitate advanced immobilization strategies to enhance their robustness and scalability. This review critically examines cutting-edge advancements in enzyme immobilization, focusing on the integration of artificial intelligence (AI), novel nanomaterials, and dynamic carrier systems to overcome the traditional limitations of mass transfer, enzyme leakage, and cost inefficiency. Key innovations such as metal–organic frameworks (MOFs), magnetic nanoparticles, self-healing hydrogels, and 3D-printed scaffolds are highlighted for their ability to optimize enzyme orientation, stability, and catalytic efficiency under extreme conditions. Moreover, AI-driven predictive modeling and machine learning emerge as pivotal tools for rationalizing nanomaterial synthesis, multi-enzyme cascade design, and toxicity assessment, while microfluidic systems enable precise biocatalyst fabrication. This review also explores emerging carrier-free strategies, including cross-linked enzyme aggregates (CLEAs) and DNA-directed immobilization, which minimize diffusion barriers and enhance substrate affinity. Despite progress, challenges persist in regards to eco-friendly nanomaterial production, industrial scalability, and real-world application viability. Future directions emphasize sustainable hybrid material design, AI-aided lifecycle assessments, and interdisciplinary synergies between synthetic biology, nanotechnology, and data analytics. By connecting laboratory innovation with industrial needs, this work provides a forward-thinking framework to harness immobilized enzymes for achieving global sustainability goals, particularly in bioremediation, bioenergy, and precision medicine. Full article

Here, we report the development of a novel bifunctional nanobiocatalyst for a one-pot cascade transformation of carboxymethyl cellulose (CMC) to D-sorbitol. The nanobiocatalyst is based on magnetic nanoparticle aggregates (MNAs) functionalized with chitosan (CS) cross-linked by tripolyphosphate (TPP). It contains two types of catalytic sites: cellulase (Cel, 5 wt.%) and Ru (3 wt.%) nanoparticles (NPs) of 0.7 nm in diameter. To optimize the nanobiocatalyst structure and composition, we first synthesized the biocatalyst, MNA-CSP-Cel (CSP stands for the CS layer cross-linked by TPP), as well as the nanocatalyst, MNA-CSP-Ru, and studied them in the one-step reactions of hydrolysis and hydrogenation, respectively. The data obtained allowed us to optimize the composition and properties of the bifunctional nanobiocatalyst, MNA-CSP-Ru-Cel, and to choose the best reaction conditions for the cascade process. MNA-CSP-Ru-Cel was characterized using transmission electron microscopy (TEM), high-resolution TEM, energy-dispersive spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and porosity measurements. The knowledge obtained enabled us to perform a cascade transformation of CMC to D-sorbitol with a yield of 83.2% for 10 h at 70 °C and a hydrogen pressure of 4 MPa. The yield demonstrated in this work is much higher than that reported to date for the same cascade process. Full article

Over the past three decades, organic reactions catalyzed by lipase have been extensively studied. To overcome the drawbacks of free enzymes and develop new and sustainable biocatalysts, various insoluble forms of lipases were examined. Especially interesting are lipases immobilized on silica nanoparticles (SiNPs) due to their promising unique and advantageous physicochemical properties. Therefore, the present paper presents an overview of different organic functionalization methods of SiNP surfaces to create a more favorable microenvironment for lipase molecules. Given the high commercial value of lipases in biotechnological applications, the second part of this paper highlights the key industrial sectors utilizing these nanobiocatalysts. This review discusses the key industrial applications of silica-based lipase nanobiocatalysts, including biodiesel production, flavor ester synthesis, and pharmaceutical applications such as racemization. Special attention is given to emerging technologies, particularly the use of immobilized lipases in polymer biodegradation and polymerization reactions. These advances have paved the way for innovative solutions, such as self-degrading bioplastics, which hold significant promise for sustainable materials and environmental protection. This comprehensive overview underscores the transformative potential of lipase–SiNP nanobiocatalysts in both industrial and environmental contexts. Full article

This study aimed to explore the capacity of immobilized lipases on the acetylation of six aglycon flavonoids, namely myricetin, quercetin, luteolin, naringenin, fisetin and morin. For this purpose, lipase B from Candida antarctica (CaLB) and lipase from Thermomyces lanuginosus (TLL) were immobilized onto the surface of ZnOFe nanoparticles derived from an aqueous olive leaf extract. Various factors affecting the conversion of substrates and the formation of monoesterified and diesterified products, such as the amount of biocatalyst and the molar ratio of the substrates and reaction solvents were investigated. Both CaLB and TLL-ZnOFe achieved 100% conversion yield of naringenin to naringenin acetate after 72 h of reaction time, while TLL-ZnOFe achieved higher conversion yields of quercetin, morin and fisetin (73, 85 and 72% respectively). Notably, CaLB-ZnOFe displayed significantly lower conversion yields for morin compared with TLL-ZnOFe. Molecular docking analysis was used to elucidate this discrepancy, and it was revealed that the position of the hydroxyl groups of the B ring on morin introduced hindrances on the active site of CaLB. Finally, selected flavonoid esters showed significantly higher antimicrobial activity compared with the original compound. This work indicated that these lipase-based nanobiocatalysts can be successfully applied to produce lipophilic derivatives of aglycon flavonoids with improved antimicrobial activity. Full article

Biodiesel is a mixture of fatty acid alkyl esters (FAAEs) mainly produced via transesterification reactions among triglycerides and short-chain alcohols catalyzed by chemical catalysts (e.g., KOH, NaOH). Lipase-assisted enzymatic transesterification has been proposed to overcome the drawbacks of chemical synthesis, such as high energy consumption, expensive separation of the catalyst from the reaction mixture and production of large amounts of wastewater during product separation and purification. However, one of the main drawbacks of this process is the enzyme cost. In recent years, nano-immobilized lipases have received extensive attention in the design of robust industrial biocatalysts for biodiesel production. To improve lipase catalytic efficiency, magnetic nanoparticles (MNPs) have attracted growing interest as versatile lipase carriers, owing to their unique properties, such as high surface-to-volume ratio and high enzyme loading capacity, low cost and inertness against chemical and microbial degradation, biocompatibility and eco-friendliness, standard synthetic methods for large-scale production and, most importantly, magnetic properties, which provide the possibility for the immobilized lipase to be easily separated at the end of the process by applying an external magnetic field. For the preparation of such effective magnetic nano-supports, various surface functionalization approaches have been developed to immobilize a broad range of industrially important lipases. Immobilization generally improves lipase chemical-thermal stability in a wide pH and temperature range and may also modify its catalytic performance. Additionally, different lipases can be co-immobilized onto the same nano-carrier, which is a highly effective strategy to enhance biodiesel yield, specifically for those feedstocks containing heterogeneous free fatty acids (FFAs). This review will present an update on the use of magnetic iron oxide nanostructures (MNPs) for lipase immobilization to catalyze transesterification reactions for biodiesel production. The following aspects will be covered: (1) common organic modifiers for magnetic nanoparticle support and (2) recent studies on modified MNPs-lipase catalysts for biodiesel production. Aspects concerning immobilization procedures and surface functionalization of the nano-supports will be highlighted. Additionally, the main features that characterize these nano-biocatalysts, such as enzymatic activity, reusability, resistance to heat and pH, will be discussed. Perspectives and key considerations for optimizing biodiesel production in terms of sustainability are also provided for future studies. Full article

In the present investigation, an ecofriendly magnetic inorganic-protein hybrid system-based enzyme immobilization was developed using partially purified laccase from Trametes versicolor (TvLac), Fe₃O₄ nanoparticles, and manganese (Mn), and was successfully applied for synthetic dye decolorization in the presence of enzyme inhibitors. After the partial purification of crude TvLac, the specific enzyme activity reached 212 U∙mg total protein⁻¹. The synthesized Fe₃O₄/Mn₃(PO₄)₂-laccase (Fe₃O₄/Mn-TvLac) and Mn₃(PO₄)₂-laccase (Mn-TvLac) nanoflowers (NFs) exhibited encapsulation yields of 85.5% and 90.3%, respectively, with relative activities of 245% and 260%, respectively, compared with those of free TvLac. One-pot synthesized Fe₃O₄/Mn-TvLac exhibited significant improvements in catalytic properties and stability compared to those of the free enzyme. Fe₃O₄/Mn-TvLac retained a significantly higher residual activity of 96.8% over that of Mn-TvLac (47.1%) after 10 reuse cycles. The NFs showed potential for the efficient decolorization of synthetic dyes in the presence of enzyme inhibitors. For up to five reuse cycles, Fe₃O₄/Mn-TvLac retained a decolorization potential of 81.1% and 86.3% for Coomassie Brilliant Blue R-250 and xylene cyanol, respectively. The synthesized Fe₃O₄/Mn-TvLac showed a lower acute toxicity towards Vibrio fischeri than pure Fe₃O₄ nanoparticles did. This is the first report of the one-pot synthesis of biofriendly magnetic protein-inorganic hybrids using partially purified TvLac and Mn. Full article

Nanobiocatalysts (NBCs) are a promising new class of biocatalysts that combine the advantages of enzymes and nanomaterials. Enzymes are biological catalysts that are highly selective and efficient, but they can be unstable in harsh environments. Nanomaterials, on the other hand, are small particles with unique properties that can improve the stability, activity, and selectivity of enzymes. The development of NBCs has been driven by the need for more sustainable and environmentally friendly bioprocessing methods. Enzymes are inherently green catalysts, but they can be expensive and difficult to recover and reuse. NBCs can address these challenges by providing a stable and reusable platform for enzymes. One of the key challenges in the development of NBCs is the immobilization of enzymes on nanomaterials. Enzyme immobilization is a process that attaches enzymes to a solid support, which can protect the enzymes from harsh environments and make them easier to recover and reuse. There are many different methods for immobilizing enzymes, and the choice of method depends on the specific enzyme and nanomaterial being used. This review explores the effective role of NBCs in pharmaceutical and biomedical fields. Full article

Enzymatic lipophilization has been proposed as a cost-effective strategy to produce new liposoluble antioxidant compounds. In this study, modified oils rich in structured phenolipids were prepared via one-pot enzymatic acylation of hydroxytyrosol (HTYR), vanillyl alcohol (VA) and homovanillyl alcohol (HVA) with pomace olive oil (POO) in solvent-free conditions using immobilized lipase on biogenic nanoparticles. The effect of temperature (30–70 °C) and enzyme concentration (0.1–1%, w/w) on the efficiency of the bioprocess as well as the reusability of the nanobiocatalyst were thoroughly investigated. The modified oils exhibited increased antioxidant activity compared to the control oil according to DPPH and CUPRAC assays (p < 0.05). The oxidative stability of pomace olive oil was also significantly enhanced after modification, as depicted by the K₂₃₂ values and TBARS contents under accelerated oxidation at 60 °C (p < 0.05). Moreover, a fortified mayonnaise containing modified oil with HTYR was prepared that was noticeably stable compared to the control mayonnaise at 28 °C for 5 months (p < 0.05). Enzymatically modified oils have great potential for application in the nutraceutical and food industry, encouraging the exploitation of immobilized lipases as effective and green catalytic tools. Full article

With advancements in bionanotechnology, the field of nanobiocatalysts has undergone rapid growth and revolutionized various nanomaterials as novel and fascinating nanocarriers for enzyme immobilization. Nanotubes, nanofibers, nanopores, nanoparticles, and nanocomposites have been successfully developed and used as nanocarriers. The construction of robust nanobiocatalysts by combining enzymes and nanocarriers using various enzyme immobilization techniques is gaining incredible attention because of their extraordinary catalytic performance, high stability, and ease of reusability under different physical and chemical conditions. Creating appropriate surface chemistry for nanomaterials promotes their downstream applications. This review discusses enzyme immobilization on nanocarriers and highlights the techniques, properties, preparations, and applications of nanoimmobilized enzymes. Full article

A biocatalyst is an enzyme that speeds up or slows down the rate at which a chemical reaction occurs and speeds up certain processes by 10⁸ times. It is used as an anticancer agent because it targets drug activation inside the tumor microenvironment while limiting damage to healthy cells. Biocatalysts have been used for the synthesis of different heterocyclic compounds and is also used in the nano drug delivery systems. The use of nano-biocatalysts for tumor-targeted delivery not only aids in tumor invasion, angiogenesis, and mutagenesis, but also provides information on the expression and activity of many markers related to the microenvironment. Iosmapinol, moclobemide, cinepazide, lysine dioxygenase, epothilone, 1-homophenylalanine, and many more are only some of the anticancer medicines that have been synthesised using biocatalysts. In this review, we have highlighted the application of biocatalysts in cancer therapies as well as the use of biocatalysts in the synthesis of drugs and drug-delivery systems in the tumor microenvironment. Full article

The present research aims to study the process of immobilization of lipase from Burkholderia cepacia by physical adsorption on graphene oxide derived (GO) from grape seed biochar. Additionally, the modified Hummers method was used to obtain the graphene oxide. Moreover, Fourier transform infrared spectroscopy, Raman spectrum, X-ray diffraction, and point of zero charge were used for the characterization of the GO. The influences of pH, temperature, enzyme/support concentration on the catalytic activity were evaluated for the immobilized biocatalyst. The best immobilization was found (543 ± 5 U/g of support) in the pH 4.0. Considering the biochemical properties, the optimal pH and temperature were 3.0 and 50 °C, respectively, for the immobilized biocatalyst. Reusability studies exhibited that the immobilized lipase well kept 60% of its original activity after 5 cycles of reuse. Overall, these results showed the high potential of graphene oxide obtained from biochar in immobilization lipase, especially the application of nanobiocatalysts on an industrial scale. Full article

In this work, we report the green production of few-layer bio-Graphene (bG) through liquid exfoliation of graphite in the presence of bovine serum albumin. Microscopic characterization evaluated the quality of the produced nanomaterial, showing the presence of 3–4-layer graphene. Moreover, spectroscopic techniques also confirmed the quality of the resulted bG, as well as the presence of bovine serum albumin on the graphene sheets. Next, for the first time, bG was used as support for the simultaneous covalent co-immobilization of three enzymes, namely β-glucosidase, glucose oxidase, and horseradish peroxidase. The three enzymes were efficiently co-immobilized on bG, demonstrating high immobilization yields and activity recoveries (up to 98.5 and 90%, respectively). Co-immobilization on bG led to an increase of apparent K_M values and a decrease of apparent V_max values, while the stability of the nanobiocatalysts prevailed compared to the free forms of the enzymes. Co-immobilized enzymes exhibited high reusability, preserving a significant part of their activity (up to 72%) after four successive catalytic cycles at 30 °C. Finally, the tri-enzymatic nanobiocatalytic system was applied in three-step cascade reactions, involving, as the first step, the hydrolysis of p-Nitrophenyl-β-D-Glucopyranoside and cellobiose. Full article

Nanobiocatalysts, i.e., enzymes immobilized on nanostructured supports, received considerable attention because they are potential remedies to overcome shortcomings of traditional biocatalysts, such as low efficiency of mass transfer, instability during catalytic reactions, and possible deactivation. In this short review, we will analyze major aspects of immobilization of cellulase—an enzyme for cellulosic biomass waste processing—on nanostructured supports. Such supports provide high surface areas, increased enzyme loading, and a beneficial environment to enhance cellulase performance and its stability, leading to nanobiocatalysts for obtaining biofuels and value-added chemicals. Here, we will discuss such nanostructured supports as carbon nanotubes, polymer nanoparticles (NPs), nanohydrogels, nanofibers, silica NPs, hierarchical porous materials, magnetic NPs and their nanohybrids, based on publications of the last five years. The use of magnetic NPs is especially favorable due to easy separation and the nanobiocatalyst recovery for a repeated use. This review will discuss methods for cellulase immobilization, morphology of nanostructured supports, multienzyme systems as well as factors influencing the enzyme activity to achieve the highest conversion of cellulosic biowaste into fermentable sugars. We believe this review will allow for an enhanced understanding of such nanobiocatalysts and processes, allowing for the best solutions to major problems of sustainable biorefinery. Full article

Commercial cellulase Cellic CTec2 was immobilized by the entrapment technique in sol–gel matrices, and sol–gel entrapment with deposition onto magnetic nanoparticles, using binary or ternary systems of silane precursors with alkyl- or aryl-trimethoxysilanes, at different molar ratios. Appropriate tailoring of the sol–gel matrix allowed for the enhancement of the catalytic efficiency of the cellulase biocatalyst, which was then evaluated in the hydrolysis reaction of Avicel microcrystalline cellulose. A correlation between the catalytic activity with the properties of the sol–gel matrix of the nanobiocatalysts was observed using several characterization methods: scanning electron microscopy (SEM), fluorescence microscopy (FM), Fourier transform infrared spectroscopy (FT-IR) and thermogravimetric analysis (TGA/DTA). The homogeneous distribution of the enzymes in the sol–gel matrix and the mass loss profile as a function of temperature were highlighted. The influence of temperature and pH of the reaction medium on the catalytic performance of the nanobiocatalysts as well as the operational stability under optimized reaction conditions were also investigated; the immobilized biocatalysts proved their superiority in comparison to the native cellulase. The magnetic cellulase biocatalyst with the highest efficiency was reused in seven successive batch hydrolysis cycles of microcrystalline cellulose with remanent activity values that were over 40%, thus we obtained promising results for scaling-up the process. Full article

The aim of this study was the biochemical and kinetic characterization of the gentisate 1,2-dioxygenase (GDO) from Pseudarthrobacter phenanthrenivorans Sphe3 and the development of a nanobiocatalyst by its immobilization on Ni²⁺-functionalized Fe₃O₄-polydopamine magnetic nanoparticles (Ni²⁺-PDA-MNPs). This is the first GDO to be immobilized. The gene encoding the GDO was cloned with an N-terminal His-tag and overexpressed in E. coli. The nanoparticles showed a high purification efficiency of GDO from crude cell lysates with a maximum activity recovery of 97%. The immobilized enzyme was characterized by Fourier transform infrared spectroscopy (FTIR). The reaction product was identified by ¹H NMR. Both free and immobilized GDO exhibited Michaelis–Menten kinetics with K_m values of 25.9 ± 4.4 and 82.5 ± 14.2 μM and V_max values of 1.2 ± 0.1 and 0.03 ± 0.002 mM·s⁻¹, respectively. The thermal stability of the immobilized GDO was enhanced at 30 °C, 40 °C, and 50 °C, compared to the free GDO. Stored at −20 °C, immobilized GDO retained more than 60% of its initial activity after 30 d, while the free enzyme completely lost its activity after 10 d. Furthermore, the immobilized nanoparticle–enzyme conjugate retained more than 50% enzyme activity up to the fifth cycle. Full article