Search Results (438)

Albendazole (ABZ) exhibits poor oral absorption; therefore, ABZ was conjugated to cholic acid to engage the apical sodium-dependent bile acid transporter (ASBT) and promote ileal uptake. ABZ–linker–CA conjugates bearing amino-alcohol linkers (C4–C8) were evaluated by integrating synthetic feasibility, purification selectivity, and ex vivo performance. Thermal aminolysis in DMF (95 °C) produced ABZ–linkers in ~50% reaction yields (HPLC-assayed), with a minor ABZ-amine by-product consistent with a workup-sensitive isocyanate route. Immobilized-lipase screening identified Lipozyme RM IM as the most effective catalyst for CA esterification in CHCl₃, showing a pronounced linker-length dependence (31% yield for C4, 25% for C6, and C8 ≤ 2.6% yield). Docking and molecular dynamics supported this trend by indicating productive binding geometries for C4/C6 but not for C8. A polarity-guided workup and silica-gel protocol enabled retrieval of unreacted intermediates and CA recycling, with cleaner separation for the C6 series. Ex vivo transport studies confirmed ASBT-mediated, linerixibat-sensitive ileal uptake, and protoscolex assays showed improved antiparasitic efficacy versus ABZ. Overall, ABZ-C6-CA offered the best balance of uptake, near-maximal efficacy, enzymatic accessibility, and separability, supporting its prioritization for scalable biocatalytic manufacturing. Full article

In this paper, batch stirred-tank alcoholysis reactions of used and refined sunflower oils were performed with n-octyl, myristyl, cetyl, oleyl, and stearyl alcohols using immobilized lipases Novozym 435 and Lipozyme IM as catalysts. Alcohol conversions ranged from 74% to 94%, with slight differences between used frying sunflower oil and refined sunflower oil. The resulting wax esters were purified via stepwise column chromatography. The different regioselectivity of the biocatalysts led to distinct reaction pathways, and Novozym 435 proved to be the most effective enzyme, providing higher conversions and no detectable by-products. This study demonstrates the valorization of waste frying oils into high-value wax esters through enzymatic alcoholysis, comparing two industrially relevant immobilized lipases and achieving high conversion across multiple long-chain alcohols. The results highlight a sustainable alternative to conventional chemical catalysis and extend biocatalytic applications beyond traditional biodiesel production. By incorporating waste lipids into value-added products, the overall sustainability and circularity of the system are improved, contributing to green and sustainable chemistry. Full article

The lipase B from Candida antarctica was immobilized on octyl-agarose using low and high (one that saturated the support surface with enzyme) loadings. Then, both biocatalysts were aminated, and the aminated and non-aminated biocatalysts were used in further experiments. The enzyme activity was determined using substrates with different structures. The modification of the four biocatalysts with poly (ethylene-alt-maleic anhydride) revealed that only a marginal covalent reaction occurs. That way, the ion exchange of the polymer on the immobilized enzyme surface should be responsible for the enzyme functional changes. The modification of the biocatalysts with this polymer produced mixed results for enzyme activity (depending on the enzyme loading, use of aminated or non-aminated enzyme, polymer concentration and used substrate), in some instances more than doubling the activity, in others reducing it by 5–6 times the activity when compared to the unmodified biocatalyst. The effects on biocatalyst stability were also mixed, depending on the same factors; in some instances, great stabilization could be found (e.g., in inactivation of the highly loaded aminated biocatalyst at pH 7.0, the unmodified biocatalyst kept 5% of the initial activity, while the biocatalyst modified with 1% of the polymer maintained 80%), but in other instances, enzyme stability was reduced after modification. It was shown that one of the effects of the polymer modification was the prevention of the enzyme release during inactivation. Full article

The present study evaluates the impact of cavitation on the performance of the chemical refining of rapeseed oils and the enzymatic interesterification of fat blends using a powerful UP400S ultrasonicator (400 W, 20 kHz). Ultrasound-assisted alkali neutralization achieved efficiency comparable to that of the conventional 60 min process in only 7 min, with similar refining losses (5.04–6.80 wt.%), although slightly higher lipid peroxidation was observed. Performing the ultrasound cavitation under a protective nitrogen atmosphere minimized the formation of lipid peroxides and their breakdown products (i.e., hexanal, nonanal), partially protected tocopherols, and improved oxidative stability (IP at 120 °C = 3.9–4.4 h). Ultrasound-assisted enzymatic interesterification (EIE) of palm kernel fat and a palm stearin blend catalyzed by immobilized lipases (Lipozyme TL IM, Lipozyme RM IM, Novozyme 435) was carried out for the first time. Cavitation accelerated triacylglycerol rearrangement, reduced reaction time from 6 h (9.0·10⁻³ to 1.6·10⁻² min⁻¹) to only 1 h (5.5·10⁻² to 1.2·10⁻¹ min⁻¹), and significantly affected melting point stabilization and solid fat content profile. In summary, ultrasound cavitation substantially enhanced mass transfer and reaction kinetics, demonstrating strong potential for process intensification in the edible oil industry. Further optimization of reaction conditions is required before large-scale industrial implementation. Full article

The accelerating global demand for sustainable energy, driven by population growth, industrialization, and environmental concerns, has intensified the search for renewable alternatives to fossil fuels. Biofuels, including bioethanol, biodiesel, biogas, and biohydrogen, offer a viable and practical pathway to reducing net carbon dioxide (CO₂) emissions. Yet, their large-scale production remains constrained by biomass recalcitrance, high pretreatment costs, and the enzyme-intensive nature of conversion processes. Recent advances in enzyme immobilization using magnetic nanoparticles (MNPs), covalent organic frameworks, metal–organic frameworks, and biochar have significantly improved enzyme stability, recyclability, and catalytic efficiency. Complementary strategies such as cross-linked enzyme aggregates, carrier-free immobilization, and site-specific attachment further reduce enzyme leaching and operational costs, particularly in lipase-mediated biodiesel synthesis. In addition to biocatalysis, nanozymes—nanomaterials exhibiting enzyme-like activity—are emerging as robust co-catalysts for biomass degradation and upgrading, although challenges in selectivity and environmental safety persist. Green synthesis approaches employing plant extracts, microbes, and agro-industrial wastes are increasingly adopted to produce eco-friendly nanomaterials and bio-derived supports aligned with circular economy principles. These functionalized materials have demonstrated promising performance in esterification, transesterification, and catalytic routes for biohydrogen generation. Technoeconomic and lifecycle assessments emphasize the need to balance catalyst complexity with environmental and economic sustainability. Multifunctional catalysts, process intensification strategies, and engineered thermostable enzymes are improving productivity. Looking forward, pilot-scale validation of green-synthesized nano- and biomaterials, coupled with appropriate regulatory frameworks, will be critical for real-world deployment. Full article

Monoacylglycerols (MAGs) are significant intermediate byproducts in the hydrolysis of oils and fats. The accumulation of MAGs not only reduces the quality and purity of the final products in biodiesel production and edible oil refining but also poses challenges for downstream separation processes. Therefore, the development of efficient biocatalysts for the specific MAG conversion is of great industrial importance. The lipase from Aspergillus oryzae (AOL) has shown potential for lipid modification; however, the wild-type enzyme (WT) suffers from poor solubility, tendency to aggregate, and low specific activity towards MAGs in aqueous systems, which severely restricts its practical application. In this study, a combinatorial protein engineering strategy was employed to overcome these limitations. We integrated fusion protein technology with rational design to enhance both the functional expression and catalytic efficiency of AOL. Firstly, the superfolder green fluorescent protein (sfGFP) was fused to the N-terminus of AOL. The results indicated that the sfGFP fusion tag significantly improved the solubility and stability of the enzyme, preventing the formation of inclusion bodies. The fusion protein sfGFP-AOL exhibited a MAG conversion rate of approximately 65%, confirming the positive impact of the fusion tag on enzyme developability. To further boost catalytic performance, site-directed mutagenesis was performed based on structural analysis. Among the variants, the mutant sfGFP-Y92Q emerged as the most potent candidate. In the MAG conversion, sfGFP-Y92Q achieved a conversion rate of 98%, which was not only significantly higher than that of sfGFP-AOL but also outperformed the widely used commercial immobilized lipase, Novozym 435 (~54%). Structural modeling and docking analysis revealed that the Y92Q mutation optimized the geometry of the active site. The substitution of Tyrosine with Glutamine at position 92 likely enlarged the substrate-binding pocket and altered the local electrostatic environment, thereby relieving steric hindrance and facilitating the access of the bulky MAG substrate to the catalytic center. In conclusion, this work demonstrates that the synergistic application of sfGFP fusion and rational point mutation (Y92Q) can dramatically transform the catalytic properties of AOL. The engineered sfGFP-Y92Q variant serves as a robust and highly efficient biocatalyst for MAG degradation. Its superior performance compared to commercial standards suggests immense potential for cost-effective applications in the bio-manufacturing of high-purity fatty acids and biodiesel, offering a greener alternative to traditional chemical processes. Full article

Background/Objectives: RELiZORB immobilized lipase cartridge (ILC) is a single-use digestive enzyme cartridge that connects in-line with enteral feeding circuits to hydrolyze triglycerides in enteral formulas. It is cleared by the FDA for pediatric and adult use. Limited data have been published regarding the effect of ILC use on growth in children younger than 5 years of age. Methods: We performed a retrospective evaluation of real-world data extracted from a third-party reimbursement program database. All patients in the program database who initiated ILC use with enteral formula when 1 to 4 years of age between 2019 and 2023 were included. Baseline and follow-up weight, height/length, and body mass index (BMI) data were collected for up to 12 months. Results: A total of 186 patients from 90 clinics in the United States were included. A subset (143 patients) with baseline and follow-up growth measurements was included in the efficacy analysis population; 76% were diagnosed with cystic fibrosis. Mean weight and BMI z-scores improved significantly (0.63 [p < 0.001] and 0.53 [p = 0.006], respectively) from baseline to 12 months after initiation of ILC use. Significant improvement in the mean weight z-score was observed after 3 months. Among people with cystic fibrosis (pwCF) who initiated ILC use when 2 to 4 years of age, those with a BMI ≥ 50th percentile increased from 22% at baseline to 43% after 12 months (p = 0.021). Improvement in weight-for-length was also observed in 1-year-old pwCF. Conclusions: Real-world evidence showed that initiation of ILC use was associated with significant improvements in mean weight and BMI z-scores among young children. Full article

This study presents a comprehensive economic and environmental evaluation of immobilized lipases produced on eggshell membrane-based carriers from eggshell waste, based on laboratory-scale experiments. By integrating economic analysis (EA) and life cycle analysis (LCA), the key factors affecting the economic viability and environmental impact of the process were identified, supporting sustainable and circular biorefinery concepts. The EA estimated the total process cost at EUR 25.63 for 15 g of product, while the effective net cost was negative (EUR −14.81) due to the valorization of anhydrous calcium chloride as a valuable by-product. The effective net cost reduction from by-product valorization of the immobilized lipase was estimated at 0.99 EUR/g as the minimum selling price (MSP). When expressed per unit of enzymatic activity, the immobilized lipase on the eggshell waste membrane-based carrier shows a substantially lower cost (EUR/U) compared with representative commercial immobilized lipases, demonstrating clear catalytic cost-efficiency advantages. The cradle-to-gate life cycle assessment, conducted using ReCiPe 2016 quantification methods, highlighted electricity consumption during drying as the primary environmental hotspot, accounting for up to 57% of the global warming potential. Sensitivity and uncertainty analyses showed that energy consumption strongly influences the impact in terms of climate change and fossil resource depletion, while the impact of chemical use was minimal. These results show that energy-efficient process optimization, especially in the drying phase, is crucial for further improving environmental and economic performance. These results indicate that optimizing energy efficiency, especially during the drying phase, is crucial for further improving the production process of immobilized lipases on eggshell membrane-based carriers, both environmentally and economically. Full article

This review surveys literature from 2010 to 2025 on Aspergillus-derived enzymes for kinetic resolution (KR), using conventional databases and AI-assisted platforms. Among over 340 species, A. niger, A. oryzae, and A. terreus are widely recognized as safe and industrially relevant. Lipases from these fungi exhibit high stability, broad substrate specificity, and enantioselectivity, enabling efficient resolution of racemic mixtures. Advances in enzyme immobilization, protein engineering, and reaction medium optimization have enhanced catalytic performance under diverse conditions. Complementary enzymes, including esterases and epoxide hydrolases, further expand biocatalytic applications. Despite increasing demand for enantiopure compounds, challenges in yield, scalability, and enzyme discovery call for integrated molecular and process strategies. Aspergillus spp. emerge as a promising system for high-level enzyme expression, offering robust secretion capacity, efficient post-translational processing, and strong adaptability for industrial biocatalysis. Full article

Ethyl butyrate is a typical flavor ester with pineapple-banana scents, but the poor yield from natural fruits limits its feasibility in food and fragrance industries. In this study, dendritic fibrous nano-silica (DFNS) was hydrophobically modified with octyl groups (DFNS-C₈) to immobilize Candida antarctica lipase B (CALB) for solvent-free esterification of ethyl butyrate. The immobilized lipase CALB@DFNS-C₈, with the enzyme loading of 354.6 mg/g and the enzyme activity of 0.064 U/mg protein, achieved 96.0% ethyl butyrate conversion under the optimum reaction conditions where the molar ratio of butyric acid to ethanol was 1:3, with a reaction temperature and time of 40 °C and 4 h. Under the solvent-free catalytic reactions, CALB@DFNS-C₈ presented the maximum catalytic efficiency of 35.1 mmol/g/h and retained 89% initial activity after ten reuse cycles. In addition, the immobilized lipase can efficiently catalyze the synthesis of various flavor esters (such as butyl acetate, hexyl acetate, butyl butyrate, etc.) and exhibits excellent thermostability and solvent tolerance. A molecular docking simulation reveals that the hydrophobic cavity around the catalytic triad stabilizes the acyl intermediate and ensures the precise orientation of both acid and alcohol substrates. This work provides new insights into the sustainable production of flavor esters using highly active and recyclable immobilized lipases through rational carrier hydrophobization and structural confinement design. Full article

The present study aimed to design a process of brown onion skin transformation by sequential extraction to a cellulose-based immobilization carrier, along with detailed analysis of obtained extracts, pointing to approaching a “zero-waste” model of circular economy. The process of brown onion skin transformation started with semi-continuous sequential subcritical extraction via consecutive use of five solvents of increasing polarity (96, 75, 50, and 25% ethanol and water), followed by alkaline liquefaction of solid residue by 10% aqueous solution of sodium hydroxide. The designed BOS transformation process resulted in 16.62 g of cellulose-based immobilization carrier derived from 100 g of brown onion skin. Extracts obtained by semi-continuous sequential subcritical extraction contained 37 mg/g of proteins, 40 mg/g of sugars, 17.5 mg/g of uronic acids, 28 mg/g of polyphenols, and 36 mg/g of flavonoids, while those obtained by alkaline liquefaction 19 mg/g of proteins, 58 mg/g of sugars, 10 mg/g of uronic acids, 6.6 mg/g of polyphenols, and 0.5 mg/g of flavonoids. The suitability of the cellulose-based enzyme immobilization carrier was evaluated by B. cepacia lipase immobilization by adsorption, where a maximal 31 U of lipase activity per 1 g of wet carrier was achieved. Based on the results obtained, it seems that the proposed process of brown onion skin transformation shows the possibility of being used for the production of a cellulose-based immobilization carrier, approaching the “zero-waste” model of a circular economy. Full article

Addressing global food waste challenges, this study investigated plant-based byproducts, spent coffee grounds, apple, and chokeberry pomaces, as sources of phenolic acids and biodegradable carriers for lipase immobilization. The goal was to enhance the lipophilicity and functionality of natural phenolics by enzymatic lipophilization. Microbial lipase from A. oryzae was immobilized on these materials, with native spent coffee grounds (NSCG) showing the highest activity (6.0 U/g hydrolytic; 1036 U/g synthetic). Chlorogenic acid (CGA), predominant in extracts, served as a model substrate. Using response-surface methodology, optimal conditions for butyl-CGA synthesis were determined. This is the first report of CGA lipophilization using food-waste-immobilized biocatalysts, where reaction yield for NSCG increased with alcohol chain length, peaking with dodecanol (34.06%). Among synthesized esters, butyl chlorogenate displayed the highest antioxidant activity, comparable to free CGA and BHT, and increased lipophilicity, though a “cut-off” effect appeared for longer chains. Medium-chain esters (C6, C8) showed selective antimicrobial activity against Gram-positive bacteria. While lipophilization of chokeberry pomace and spent coffee grounds extracts reduced antioxidant activity, short-chain esters (C4–C6) improved rapeseed oil stability. The findings highlight food waste as a sustainable source for developing biocatalysts and value-added bioactives with enhanced functional properties. Full article

Objective: This study aimed to develop a stable and active biocatalytic system for enzyme immobilization, utilizing an electrospun support doped with a metal–organic framework (MOF) and supplemented with an ionic liquid as a lipase stabilizer and activity enhancer. Methodology: The system was applied for an efficient and enantioselective resolution of racemic citalopram. Key parameters, including MOF concentration, electrospinning and immobilization conditions, ionic liquid selection, and reaction time, were optimized to enhance biocatalyst performance. Results: The optimal immobilization time was determined to be 24 h, achieving 52% immobilization efficiency and 100% activity recovery. The resulting biocatalytic system HIGH PVC-MOF-lip-CA exhibited superior storage stability, retaining 80% of its initial activity, a 75% improvement over the free enzyme. In the resolution of citalopram, the system achieved 96% conversion of S-citalopram within 24 h, with the enantiomeric excess of 93% in favor of the S-ester over the R-ester. These findings demonstrate the system’s potential for efficient and stereoselective biocatalytic applications. Full article

Saturated and branched high molecular weight organic esters are highly valued as emollients in the cosmetic industry due to their superior properties. Their saturated character provides resistance to oxidation and rancidity. Additionally, their branched structure endows them with low melting temperatures, enabling them to remain liquid over a broad temperature range. These esters can be obtained from branched alcohols, branched fatty acids or both, using chemical or enzymatic processes. Among branched alcohols, Guerbet alcohols stand out. Due to their characteristic properties as branched, saturated alcohols with superior oxidative stability and extremely low volatility, they are proposed as excellent substrates for the enzymatic synthesis of these compounds. This study represents the first investigation into the biocatalytic synthesis of three specific esters: those formed between 2-octyl-1-dodecanol (C20 Guerbet alcohol) and the fatty acids myristic (MA), palmitic (PA), and stearic acid (SA). To achieve this, an environmentally sustainable biocatalytic process was developed. The synthesis involves a solvent-free esterification catalyzed by the commercial immobilized lipase, Lipozyme^® 435, conducted within a vertically stirred, thermostated batch tank reactor. Optimal conditions for lipase concentration and temperature were established, and the sustainability of the process was successfully quantified using various “green metrics”. Full article

Lipases (EC 3.1.1.3) catalyze the hydrolysis of triacylglycerols into mono- and diacylglycerols and free fatty acids. This study investigated the production of lipase by Hypocrea pseudokoningii under solid-state fermentation (SSF), followed by its immobilization, purification, and biochemical characterization. Maximum activity was achieved using wheat fiber after 168 h of cultivation. Supplementation with oils enhanced production, particularly palm oil (315U; 1.58-fold increase) and soybean oil (Glycine max) (298U; 1.49-fold increase). The addition of micronutrients further improved yields, with Khanna (445U) and Vogel (400U) salts promoting more than a two-fold increase in activity. Immobilization on Octyl-Sepharose significantly altered the enzyme’s properties. The free lipase exhibited optimal activity at 45 °C and pH 4.5–5.5, while the immobilized enzyme showed maximum activity at 35–40 °C and pH 5.5. Thermal stability was notable enhanced: the free lipase had a half-life of 10 min at 50 °C, whereas the immobilized enzyme remained stable for 60 min and retained over 30% activity at 70 °C. Both the free and immobilized forms were stable across a broad pH range (4.0–10.0), maintaining more than 70% residual activity. The enzyme was stabilized by Tween 80 but inhibited by SDS. It was activated by Ca²⁺ and showed resistance to Pb²⁺, Zn²⁺, and Cu²⁺. Hydrolytic assays revealed murumuru (Astrocaryum murumuru), cupuaçu (Theobroma grandiflorum), and soybean oils as preferred substrates. TLC confirmed the formation of mono- and diglycerides, as well as the presence of fatty acids. Full article