Search Results (88)

Proteases are key biocatalysts widely applied in the food, pharmaceutical, detergent, and environmental industries. One of the most costly steps in large-scale enzyme production is the preparation of the culture medium, making agro-industrial wastes attractive as low-cost nutrient sources and potential inducers. The non-conventional yeast Yarrowia lipolytica stands out in bioprocess engineering due to its high secretion capacity, GRAS status, and ability to metabolize diverse industrial residues. In this study, Brazilian agro-industrial by-products, namely Corn steep liquor (CSL), brewer’s yeast residue (BYR), and okara, were evaluated as alternative nitrogen sources for protease production by Y. lipolytica IMUFRJ 50678. Enzyme activity was quantified by the azocasein method at optimized conditions (40 °C, 40 min, pH 5 and 8). After an initial exploratory screening (n = 1), brewer’s yeast residue (BYR) and okara were identified as promising candidates for protease production. These preliminary findings guided subsequent experiments performed in biological triplicate (n = 3), which confirmed the reproducibility and comparative performance of these substrates, showing higher acid protease (AXP) activity in the BYR medium ((5.4 ± 0.3) U/mL), whereas alkaline protease (AEP) activities were comparable between the BYR ((8.4 ± 0.6) U/mL) and okara ((7.5 ± 0.9) U/mL) media. CSL was associated with higher lipase activity ((11.7 ± 0.9) × 10³ U/L), while esterase activity was higher in the BYR medium. These findings indicate that agro-industrial residues, particularly BYR and okara, can serve as effective nitrogen sources for protease production by Y. lipolytica IMUFRJ 50678, supporting their use in waste valorization and sustainable bioprocesses. Full article

Agro-industrial waste impacts populations worldwide. Food waste, in turn, is a major source of complex lipids, carbohydrates, and other substances. Therefore, it is crucial to convert food waste into products that reduce environmental problems. Enzymatic hydrolysis has advantages over chemical hydrolysis. Examples include the enzymatic hydrolysis of starch by α-amylase and the hydrolysis of inulin by inulinase, which occur under milder environmental and temperature conditions than acid hydrolysis of starch or inulin. Despite these milder temperature conditions, during substate hydrolysis, enzyme deactivation occurs under exposure to temperature. As temperature increases above $T_{opt}$ (which maximizes catalytic activity), enzyme deactivation becomes more pronounced, leading to a decrease in enzyme activity. Therefore, determining the rate constant of deactivation $k_d$, during biotransformation is an important aspect in understanding enzyme kinetics. Most experimental studies focus on changes in enzyme activity with time and temperature. However, enzyme deactivation also occurs during enzymatic reactions conducted at different temperatures, and this process is characterized by specific deactivation parameters. The study is to present the rate constants of deactivation $k_d$, for selected biotransformation processes. The selected biotransformation processes are hydrolysis of olive oil by lipase, hydrolysis of inulin by inulinase, and hydrolysis of starch by α-amylase. Given the widespread use of enzymes in industry, the information on enzyme deactivation presented in this study can be used by engineers involved in modeling and optimizing enzymatic processes. This knowledge is also essential for the effective and sustainable use of enzymes in industrial applications. It is important to emphasize that the deactivation parameters discussed in this study also carry significant economic, social, and environmental implications. 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

The global change toward sustainable manufacturing has intensified the development of alternatives to petrochemical-based surfactants, which are environmentally recalcitrant and fossil dependent. Biosurfactants have emerged as the most promising petrochemical-based surfactant substitutes, due to their biodegradability, low toxicity, and robust performance under extreme conditions; however, their industrial use is hindered by high production costs, limited productivity, and complex downstream processing, for instance high protein content can make the ultrafiltration (downstream strategy) unfeasible. This review critically examines recent advances in integrated bioprocess design to overcoming these constraints, with particular emphasis on the convergence of enzymatic catalysis and microbial fermentation. Comparative assessment across key biosurfactant classes demonstrates that tailored enzymatic transformations, enabled by lipases, glycosyltransferases, acyltransferases, and oxidoreductases, offer precision in structural modification unattainable through fermentation alone, enabling programmable amphiphilicity and improved functional performance. Thus, the translation of enzymatic and hybrid routes to industry remains restricted by enzyme stability, cofactor regeneration, and process engineering challenges. Emerging strategies such as continuous fermentation, in situ product recovery, and machine learning-based process control show strong potential to enhance productivity and reduce energy demands. By integrating molecular design, metabolic engineering, and intensified bioprocessing, this review delineates a strategic framework for advancing next-generation biosurfactants toward commercial viability within circular and sustainable value chains. 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

The rapid accumulation of plastic and textile waste, particularly polyethylene terephthalate (PET), has emerged as a global challenge for sustainable resource management. Conventional recycling methods, including mechanical and chemical routes, recover limited value and often degrade material quality while consuming substantial energy. Biocatalytic recycling, by contrast, offers a resource-efficient alternative that transforms post-consumer PET into high-purity monomers under mild and environmentally benign conditions. This review examines advances in enzymatic PET depolymerization, focusing on hydrolases such as cutinases, PETases, MHETases, and lipases. The discussion highlights enzyme engineering, reactor design, and process integration that improve kinetics, thermostability, and yield. From a resource perspective, biocatalytic recycling redefines PET waste as a renewable carbon feedstock capable of re-entering industrial cycles, thereby reducing reliance on virgin petrochemicals and mitigating greenhouse gas emissions. Ultimately, this review positions biocatalytic PET recycling as a cornerstone technology for achieving circularity and advancing global resource sustainability. Full article

The design and development of scaffolds play a crucial role in tissue engineering. In this regard, the study aims to establish the influence of porosity on the morpho-structural, physical–chemical, and biochemical characteristics of the polylactic acid (PLA) and/or polycaprolactone (PCL) scaffolds, in order to be considered candidates for tissue reconstruction. The results indicated that binary PLA-PCL and PCL matrices are more suitable than PLA, due to their higher crystallization degree, this contributing to the superior mechanical properties and lower network defects. The preponderance of molecular interactions decreases with porosity. Porosity induced a decrease in the degree of crystallization of PLA-PCL and an increase in water, glucose and blood components uptake by 188, 178, and 28%, respectively. The PLA-PCL scaffold was found to be more stable to lipase action than neat PLA as a result of the reduced enzyme access due to the higher crystallinity and thermodynamic stability of the hydrocarbon linear chain in PCL, which is higher than that of the side methyl group in PLA. Lactobacillus growth increases with porosity and was more pronounced on the PLA-PCL matrix. All these results show that varying the porosity and composition of the polymer mixture leads to valuable materials with nutrient absorption capacity and biodegradability superior to neat PLA or PCL materials. Full article

Chronic pancreatitis (CP) precipitates complex malnutrition through synergistic mechanisms: exocrine pancreatic insufficiency–driven maldigestion, duodenal or pancreatobiliary strictures limiting nutrient flow, cholestasis impairing micelle formation, alcohol-related anorexia, pain-induced hypophagia, proteolytic catabolism from type 3c diabetes, and a chronic inflammatory milieu that accelerates sarcopenia and bone demineralisation. Consequent calorie–protein depletion, micronutrient and fat-soluble vitamin deficits, and metabolic derangements markedly amplify morbidity. Pancreatic enzyme replacement therapy (PERT) with targeted micronutrient repletion is foundational; high-protein regimens co-administered with PERT curb muscle loss, and medium-chain triglycerides (MCTs) can augment caloric delivery by bypassing lipase dependence, although their benefit over personalised dietetic counselling is marginal. Optimal dietary fat thresholds and timing of escalation from oral to enteral or parenteral feeding remain unresolved. Comprehensive care also demands alcohol abstinence, effective analgesia and stringent glycaemic control. Serial monitoring—biochemical indices, densitometry, dual-energy X-ray absorptiometry and imaging-based body-composition metrics—permits early detection of high-risk patients and precision tailoring of interventions. Intensified multidisciplinary programmes already improve prognostic endpoints and are unveiling biomarkers of nutritional resilience. A structured, evidence-based strategy integrating PERT, macronutrient engineering, micronutrient repletion and metabolic surveillance is essential to mitigate nutrition-related morbidity, enhance long-term outcomes and optimise quality of life in CP. Full article

As the supply of cocoa becomes increasingly volatile, biotechnological innovations such as lipid engineering with lipases play a crucial role in supporting more stable, ethical, and sustainable chocolate production systems. This study explores the potential of Pseudomonas fluorescens lipase immobilized on eggshell membrane-based carriers for the synthesis of a cocoa butter substitute (CBS). The carriers were prepared by treating eggshells with different acids to generate chemically distinct support materials. Lipase immobilization was performed using both adsorption and covalent binding techniques. All resulting biocatalysts were characterized and compared to the free enzyme with respect to pH and temperature optima, as well as thermal and solvent stability. Immobilization caused shifts in the enzyme’s optimal operating conditions and significantly improved its stability at elevated temperatures and in the presence of organic solvents. Among the tested systems, the lipase immobilized by adsorption onto a hydrochloric acid-treated carrier exhibited the best performance. Using this biocatalyst, a CBS containing 93.54 ± 0.16% of the target triacylglycerols (POP, POS, and SOS) was successfully synthesized and reused over five consecutive synthesis cycles without significant loss of activity. These findings demonstrate the potential of waste-derived biomaterials for the development of efficient, stable, and reusable biocatalysts in the enzymatic production of functional lipids. Full article

Poly(ε-caprolactone) (PCL) is a synthetic plastic known for its excellent physicochemical properties and a wide range of applications in packaging, coatings, foaming, and agriculture. In medicine, its versatility allows it to function as a scaffold for drug delivery, sutures, implants, tissue engineering, and 3D printing. In addition to its biocompatibility, PCL’s most notable characteristic is its biodegradability. However, this property is affected by temperature, microbial activity, and environmental conditions, which means PCL can sometimes remain in nature for long periods. This review shows that various types of microorganisms can efficiently degrade PCL, including different strains of Pseudomonas spp., Streptomyces spp., Alcaligenes faecalis, and fungi like Aspergillus oryzae, Fusarium spp., Rhizopus delemar, and Thermomyces lanuginosus. These microorganisms produce enzymes such as lipases, esterases, and cutinases that break down PCL into smaller molecules that act as substrates. The review also examines the phylogenetic diversity of organisms capable of biodegrading PCL, the biochemical pathways involved in this process, and specific aspects of the genetic framework responsible for the expression of the enzymes that facilitate degradation. Targeted research on microbial PCL biodegradation and its practical applications could significantly aid in reducing and managing plastic waste on a global ecological scale. Full article

Olive oil production generates vast quantities of by-products, with olive mill wastewater (OMW) being a particularly challenging effluent. Characterized by its dark color, high acidity, and rich composition of organic matter, phenolic compounds, and residual oils, OMW resists conventional degradation methods and poses significant environmental risks due to its phytotoxicity and microbial inhibition. Addressing this issue requires sustainable solutions that align with circular economy principles. A promising strategy involves the biotechnological valorization of OMW using the non-conventional yeast Yarrowia lipolytica, which thrives on organic-rich substrates and converts them into high-value metabolites. This review provides a comprehensive analysis of recent advances in Y. lipolytica applications for OMW valorization, emphasizing its role in developing eco-friendly industrial processes. It begins by outlining the physicochemical challenges of OMW and the metabolic versatility of Y. lipolytica, including its ability to adapt to acidic, phenolic-rich environments. Subsequent sections critically evaluate the yeast’s capacity to synthesize commercially valuable products such as lipases (used in the food and biofuel industries), citric acid (a food and pharmaceutical additive), and polyols like mannitol and erythritol (low-calorie sweeteners). Strategies to optimize microbial productivity, such as substrate pre-treatment, nutrient supplementation, and process engineering, are also discussed. By synthesizing current research, the review highlights how Y. lipolytica-driven OMW valorization can mitigate environmental harm while creating economic opportunities, bridging the gap between waste management and green chemistry. Full article

The site-directed immobilization of enzymes has demonstrated significant potential in industrial applications due to its ability to minimize enzyme heterogeneity and maximize retained activity. However, existing approaches often require the introduction of unnatural amino acids or excessive specific ligase to achieve this goal. In this study, a self-catalyzed protein capture system (i.e., the SnoopCatcher/SnoopTag pair) was utilized for the directed immobilization of lipase on magnetic carriers. By tagging the Pseudomonas fluorescens lipase (PFL) with a SnoopTag at the C-terminal, the fused lipase PFL-SnoopTag (PSNT) readily conjugated with the SnoopCatcher partner via a spontaneously formed isopeptide bond between them. Novel magnetic particles functionalized by SnoopCatcher proteins were prepared using a co-precipitation method, achieving a loading capacity of around 0.8 mg/g carrier for the SnoopCatcher. This functionalized magnetic carrier enabled the site-directed immobilization of lipase PSNT at 81.4% efficiency, while the enzyme loading capacity reached 3.04 mg/g carriers. To further assess the practical performance of site-directed immobilized lipases, they were applied in biodiesel production and achieved a yield of 88.5%. Our results demonstrate a universal platform for the site-directed immobilization of enzymes with high performance, which offers significant advantages, e.g., single-step purification and catalyst-free immobilization of engineered enzymes, as well as easy recovery, highlighting its potential for industrial applications. Full article

This study aims to delve into the application potential of immobilized lipases in the catalytic synthesis of isoamyl acetate. Through a comparative analysis of various immobilization methods, including physical adsorption, encapsulation, covalent binding, and crosslinking, along with the utilization of nanomaterials, such as magnetic nanoparticles, mesoporous silica SBA-15, and covalent organic frameworks (COFs) as carriers, the study systematically evaluates their enhancing effects on lipase catalytic performance. Additionally, solvent engineering strategies, encompassing the introduction of organic solvents, supercritical fluids, ionic liquids, and deep eutectic solvents, are employed to intensify the enzymatic catalytic process. These approaches effectively improve mass transfer efficiency, activate enzyme molecules, and safeguard enzyme structural stability, thereby significantly elevating the synthesis efficiency and yield of isoamyl acetate. Consequently, this research provides solid scientific rationale and technical support for the industrial production of flavor ester compounds. Full article

Lipases are enzymes that catalyze the hydrolysis of carboxylic esters at a lipid–water interface and are able to catalyze reactions such as alcoholysis, esterification, transesterification, and enantioselective synthesis in organic media. They are important biocatalysts for biotechnological and industrial applications—such as in the food and flavor industry—and in the production of biopharmaceuticals, biofuels, biopolymers, and detergents. A desirable property of lipases is stereoselectivity for the production of chemicals with high optical purity. In this work, we report the improvement of the enantioselective capabilities of the Geobacillus thermoleovorans CCR11 lipase. By means of a rational design and bioinformatic approaches, six amino acids of the catalytic cavity of the lipase LipTioCCR11 were substituted resulting in an increase in the optimum temperature of the enzyme and in the resistance to the presence of organic solvents in hydrolytic reactions, and in the promotion of the enantioselective recognition of R isomers of carboxylic acids with importance for the pharmaceutical and food industries. Full article