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Aquaculture is one of the fastest-growing food sectors, producing almost 50% of fish for human consumption worldwide. However, relying on fish meal and oil for aquaculture diets is not sustainable economically or environmentally. Aquaculture feeds also contain terrestrial plant ingredients with indigestible forms of phosphorus (P), of which 70%–80% can be released into aquatic environments. This P influx contributes to eutrophication of freshwater ecosystems that can lead to anoxic conditions. This study explores a more sustainable diet for salmonids, an important and valuable seafood. Our aim was to test ingredients with highly digestible forms of P in nutritionally balanced portions to support fish growth and reduce P loading. We determined the digestibility of three feeds containing raw, extruded, and enzymatically processed microalgal co-product of Nannochloropsis oculata compared to a conventional diet. We also quantified how much P was retained and excreted. We detected highest growth in trout fed enzymatically processed co-product feed, though it was not statistically different (p = 0.846) from growth of fish fed the reference or other co-product diets. The enzyme-treated, microalgal co-product ingredient and diet had comparable values for P digestibility and solid P excretion to the reference diet, but the lowest average solid P excretion of all test diets. Trout fed the enzyme-treated diet had the highest P retention, while the reference diet had the lowest (p = 0.0429). Trout fed the enzyme-treated diet had the lowest (p = 0.0174) and negative dissolved P excretion, while those fed the reference diet had the highest. Results showed that enzyme-treated N. oculata co-product maintains digestibility, increases P retention, and reduces dissolved P excretion compared to the reference diet in rainbow trout. These findings encourage follow-up research to design and test growth performance of diets containing enzyme-treated microalgal co-product as sustainable trout aquafeed.

The triazole antifungals posaconazole and itraconazole can cause pseudohyperaldosteronism with hypertension and hypokalemia, edema, and gynecomastia by inhibiting steroid synthesis and metabolism. Mechanisms underlying pseudohyperaldosteronism include inhibition of adrenal 11β-hydroxylase cytochrome-P450 (CYP) 11B1 and 17α-hydroxylase (CYP17A1) as well as peripherally expressed 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2). To enhance specificity for fungal CYP51, tetrazoles have been developed. This study employed H295R adrenocortical cells and enzyme activity assays to assess the potential risk of oteseconazole and two other tetrazoles, VT-1598 and quilseconazole, to inhibit adrenal steroidogenesis or 11β-HSD2. Steroidomic footprint analyses of H295R cell supernatants using untargeted liquid-chromatography-high-resolution mass-spectrometry (LC-HRMS) indicated overall patterns common to oteseconazole, quilseconazole and itraconazole, as well as similarities between VT-1598 and isavuconazole. Additionally, more specific features of the steroid signatures were observed. Targeted quantification of nine adrenal steroids in supernatants from treated H295R cells revealed an overall inhibition of adrenal steroidogenesis by the three tetrazoles, itraconazole and isavuconazole, providing an explanation for their similar steroidomic pattern. Applying recombinant enzymes indicated that this effect is not due to direct inhibition of steroidogenic enzymes because no or only weak inhibition could be observed. Moreover, oteseconazole and the two other tetrazoles did not inhibit 11β-HSD2, suggesting that they do not pose a risk of pseudohyperaldosteronism. Furthermore, oteseconazole did not alter steroid concentrations in a recent clinical study. Nevertheless, follow-up studies should assess the mechanism underlying the observed overall steroidogenesis inhibition by tetrazoles, itraconazole and isavuconazole, and whether concentrations achievable in a subgroup of susceptible patients might cause adrenal insufficiency and hyperplasia.

Chitin is an abundant biopolymer and pathogen-associated molecular pattern that stimulates a host innate immune response. Mammals express chitin-binding and chitin-degrading proteins to remove chitin from the body. One of these proteins, Acidic Mammalian Chitinase (AMCase), is an enzyme known for its ability to function under acidic conditions in the stomach but is also active in tissues with more neutral pHs, such as the lung. Here, we used a combination of biochemical, structural, and computational modeling approaches to examine how the mouse homolog (mAMCase) can act in both acidic and neutral environments. We measured kinetic properties of mAMCase activity across a broad pH range, quantifying its unusual dual activity optima at pH 2 and 7. We also solved high-resolution crystal structures of mAMCase in complex with oligomeric GlcNAcn, the building block of chitin, where we identified extensive conformational ligand heterogeneity. Leveraging these data, we conducted molecular dynamics simulations that suggest how a key catalytic residue could be protonated via distinct mechanisms in each of the two environmental pH ranges. These results integrate structural, biochemical, and computational approaches to deliver a more complete understanding of the catalytic mechanism governing mAMCase activity at different pH. Engineering proteins with tunable pH optima may provide new opportunities to develop improved enzyme variants, including AMCase, for therapeutic purposes in chitin degradation.

There are many occurrences of enzymes catalysing the same reaction but having significantly different structures. Leveraging the comprehensive information on enzymes stored in the Mechanism and Catalytic Site Atlas (M‐CSA), we present a collection of 34 cases for which there is sufficient evidence of functional convergence without an evolutionary link. For each case, we compare enzymes which have identical Enzyme Commission numbers (i.e. catalyse the same reaction), but different identifiers in the CATH data resource (i.e. different folds). We focus on similarities between their sequences, structures, active site geometries, cofactors and catalytic mechanisms. These features are then assessed to evaluate whether all the evidence for these structurally diverse proteins supports their independent evolution to catalyse the same chemical reaction. Our approach combines published literature information with knowledge‐based computational resources from, amongst others, M‐CSA, PDBe and PDBsum, supported by tailor‐made software to explore active site structures and assess similarities in mechanism. We find that there are multiple types of convergent functional evolution observed to date, and it is necessary to investigate sequence, structure, active site geometry and enzyme mechanisms to describe such convergence accurately. [ABSTRACT FROM AUTHOR]

Live‐cell imaging of RNA in specific subcellular compartments is essential for elucidating the rich repertoire of cellular functions, but it has been limited by a lack of simple, precisely controlled methods. Here such an approach is presented via the combination of hybridization chain reaction and spatially restricted enzymatic activation with organelle‐targeted delivery. The system can localize engineered DNA hairpins in the mitochondria, where target RNA‐initiated chain reaction of hybridization events is selectively activated by a specific enzyme, enabling amplified RNA imaging with high precision. It is demonstrated that the approach is compatible with live cell visualization and enables the regulatable imaging of microRNA in mitochondria. Since in situ activation of the signal amplification with enzyme eliminates the need for genetically encoded protein overexpression, it is envisioned that this simple platform will be broadly applicable for precise RNA imaging with subcellular resolution in a variety of biological processes. [ABSTRACT FROM AUTHOR]

To investigate the effect of nitrogen fertilizer and zinc (Zn) sulfate on growth, physiological, biochemical, and nutrient use efficiency in fodder maize under drought stress, an experiment was conducted during two cropping years 2019 and 2020. The experiment was performed as a split-factorial in the form of a randomized complete block design with four replications. The main factor was irrigation regimes in three levels: full irrigation (without stress or 100% of field capacity), irrigation based on 75% of field capacity, and irrigation based on 50% of field capacity. The first sub-factor included urea fertilizer at three levels of 0, 69, and 138 kg ha−1. The second sub-factor included foliar application of zinc sulfate at three levels of 0, 2, and 4 mg kg−1. The results showed that the highest fresh weight of maize fodder was obtained under conditions of full irrigation and the use of 138 kg ha−1 of N. The use of zinc at all irrigation levels increased the fresh weight of maize fodder. These results confirmed that if higher amounts of N are used, more zinc should be used to achieve a higher yield of maize forage. With the increase in the severity of drought stress, the concentration of nitrogen and phosphorus in the crop tissue, as well as the efficiency of using nitrogen and phosphorus, decreased. The highest value of nitrogen use efficiency was obtained in full irrigation conditions, using 69 kg ha−1 of N and 4 mg kg−1 of zinc. In addition, at all irrigation levels, the efficiency of using this element decreased with increasing zinc consumption. [ABSTRACT FROM AUTHOR]

This study focuses on incorporating liquid molecules, different from the bulk solution, into the immobilized enzyme carrier to regulate the distribution effect and diffusion‐limiting impact of the carrier's microenvironment for substrates, which generally is difficult to achieve due to the instability of the materials with liquid inclusions. A freestanding liquid‐holding particle carrier, with bi‐component hydrophobic ionic liquid gel microspheres with poly (glycidyl methacrylate) as the network and 1‐butyl‐3‐methylimidazolium hexafluorophosphate as the dispersing medium, is proposed, which is stable in air aqueous solution, and can extract proteins and organic small molecules into its interior due to the mobility of its dispersing medium. Horseradish peroxidase is covalently immobilized into the microspheres, forming a liquid compartment enzyme microreactor. The microreactor exhibits superior stability, enzymatic activity, and catalytic performance for Basic Orange II degradation compared to free enzyme and liquid‐free immobilized enzymes. This is attributed to the biocompatibility of the ionic liquid, its role in substrate enrichment in its interior, and its rapid mass transfer capability. This contribution shows the effectiveness of regulating the carrier's microenvironment with liquid molecules, offering fresh perspectives and strategies for enzyme technology. [ABSTRACT FROM AUTHOR]

Recent research has looked at various macroalgae species as dietary components or feed additives for a variety of fish species due to their nutritional value. The objective of this study was to examine the impact of Najas indica, a macroalgae extract, on the growth performance, proximate composition, and metabolic activities of monosex tilapia (Oreochromis niloticus), while also isolating the compounds present. Three distinct solvents (n‐hexane, ethyl acetate, and ethanol) were used to extract bioactive compounds from a coarse powder of macroalgae after drying and grinding, and gas chromatography–mass spectrometry (GC–MS) analysis was used to detect bioactive compounds. The extracts were combined with commercial feed (0.4%) and applied to the treatment with three replications and a control containing 50 fingerlings per tank for 5 weeks. The findings indicated a significant increase in the final weight, weight gain, specific growth rate (SGR), and survival rate among the treated fish, whereas the feed conversion ratio (FCR) was observed to decrease in comparison to the control group. Significantly higher levels of protein and lipids were found in treated fish, whereas moisture and ash levels were significantly lower compared to control fish. In treated fish, the digestive enzyme amylase was significantly higher, but the protease enzyme reduced significantly. The antioxidant enzyme superoxide dismutase activity (SOD) was significantly higher in the treatment group, whereas the catalase (CAT) enzyme did not differ significantly. A total of 47 bioactive compounds were identified in N. indica, among which the prominent compounds included n‐hexadecanoic acid, neophytadiene, phytyl palmitate, d‐mannitol, and heptanoic acid. The results obtained from this study indicate that the utilization of N. indica macroalgae extract has the potential to serve as an additional dietary component, therefore, enhancing the growth performance and metabolic functions of fish. [ABSTRACT FROM AUTHOR]

Colon cancer is on the rise in younger adults. Despite multimodal treatment strategies, clinical outcomes in advanced stage colon cancer patients remain poor. Neoadjuvant/adjuvant chemotherapy efficacy is limited due to chemoresistance, toxicity, and negative side effects. Overwhelming evidence supporting the small‐molecule metabolites derived from breakdown of food or microbial sources confer an extensive array of host benefits, including chemo‐preventive role in colon cancer. Our previous study indicated that the introduction of glyoxylate (Glx), an intermediate product of microbial or plant metabolism, exerts a cytotoxic effect in colon cancer cells. This study was designed to evaluate the effects of Glx on colon cancer with molecular insights. For this, we established an AOM/DSS‐induced colitis associated colon cancer model in mice. Supplementation of Glx in vivo reduced colitis associated tumor growth and altered the metabolic characteristics of tumor tissue in mice without initiating any severe liver or renal toxicity. More specifically, intake of glyoxylate accumulated glycine in the colon tissue by elevation of alanine‐glyoxylate transferase (AGXT) activity. Glycine accumulation increased intracellular Ca2+ concentration via glycine receptor activation and dysregulation of Ca2+ homeostasis lead to induction of apoptosis that resulted in arresting tumor growth. Interestingly, elevation of AGXT activity or Glx related specific metabolic pathway provides better survival in colon cancer patients. Collectively, our exclusive findings support the exploration of Glx either as a preventive molecule or its inclusion in the treatment regimens for colon cancer. [ABSTRACT FROM AUTHOR]

Background Glycated hemoglobin, or hemoglobin A1c (HbA1c), serves as a crucial marker for diagnosing diabetes and monitoring its progression. We aimed to assess the interference posed by common Hb variants on popular HbA1c measurement systems. Methods A total of 63 variant and nonvariant samples with target values assigned by the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC)-calibrated methods were included. We assessed 6 methods for measuring HbA1c in the presence of HbS, HbC, HbD, HbE, and fetal hemoglobin (HbF): 2 cation-exchange high-performance liquid chromatography (HPLC) methods (Bio-Rad D-100 and HLC-723 G8), a capillary electrophoresis (CE) method (Sebia Capillarys 3 TERA), an immunoassay (Roche c501), an enzyme assay system (Mindray BS-600M), and a boronate affinity method (Primus Premier Hb9210). Results The HbA1c results for nonvariant samples from the 6 methods were in good agreement with the IFCC-calibrated method results. The Bio-Rad D-100, Capillarys 3, Mindray BS-600M, Premier Hb9210, and Roche c501 showed no interference from HbS, HbC, HbD, and HbE. Clinically significant interference was observed for the HLC-723 G8 standard mode. Elevated HbF levels caused significant negative biases for all 6 methods, which increased with increasing HbF concentration. Conclusion Elevated levels of HbF can severely affect HbA1c measurements by borate affinity, immunoassays, and enzyme assays. [ABSTRACT FROM AUTHOR]

Conventional chemotherapeutic agents are limited by their lack of targeting and penetration and their short retention time, and chemotherapy might induce an immune suppressive environment. Peptide self‐assembly can result in a specific morphology, and the resulting morphological changes are stimuli responsive to the external environment, which is important for drug permeation and retention of encapsulated chemotherapeutic agents. In this study, a polypeptide (Pep1) containing the peptide sequences PLGLAG and RGD that is responsive to matrix metalloproteinase 2 (MMP‐2) was successfully developed. Pep1 underwent a morphological transformation from a spherical structure to aggregates with a high aspect ratio in response to MMP‐2 induction. This drug delivery system (DI/Pep1) can transport doxorubicin (DOX) and indomethacin (IND) simultaneously to target tumor cells for subsequent drug release while extending drug retention within tumor cells, which increases immunogenic cell death and facilitates the immunotherapeutic effect of CD4+ T cells. Ultimately, DI/Pep1 attenuated tumor‐associated inflammation, enhanced the body's immune response, and inhibited breast cancer growth by combining the actions of DOX and IND. Our research offers an approach to hopefully enhance the effectiveness of cancer treatment. [ABSTRACT FROM AUTHOR]

• The content of Melilotus officinalis was revealed by LC-MS/MS analysis. • A novel potentiometric sensor detected antioxidant activity levels in extracts. • Active sites of the most abundant molecules were shown by molecular docking. Conducting content analysis is very important to elucidate and expand the different usage areas of plant components. Today's content analysis methods provide very reliable results. However, the high cost of these methods and the need for trained personnel bring other challenges. Therefore, simple, fast, and low-cost content analysis methods are a great need. In this study, the phenolic content of Melilotus officinalis flower extracts was investigated using LC-MS/MS analysis, and its antioxidant activity was determined with a novel method employing a PVC membrane sensor. In the method, for the first time, the antioxidant activities of M. officinalis flowers were determined in an economical, sensitive, fast, and stable manner using a PVC potentiometric sensor. Sensor measurements were performed using DPPH and FCR molecules, resulting in 25.71 mg.g-1 activity values and 117.524 mg.g-1, respectively. LC-MS/MS analysis was conducted to investigate the presence of 45 standard phenolic compounds in M. officinalis. The results revealed that coumarin and isoquercitrin were the major phenolic compounds in the M. officinalis flower extract. The findings of LC-MS/MS analyses of the extracts show moderate phenolic content presence in the M. officinalis flower extracts. The total phenolic content determined by LC-MS/MS analysis was consistent with the PVC membrane sensor analysis results. The studies of molecular docking aimed to identify the most effective active sites, and the Moldock scores (-57.13, -105.93) and binding energies (4.5 kcal.mol-1, -7.9 kcal.mol-1) of coumarin and isoquercitrin molecules. The results of molecular docking studies suggest that isoquercitrin exhibits a stronger inhibitory effect on DD-peptidase enzymes compared to coumarin. [ABSTRACT FROM AUTHOR]

Rationale: The compound 5-((4-methoxyphenyl)thio)benzo[c][1,2,5]thiadiazole (MTDZ) has recently been shown to inhibit in vitro acetylcholinesterase activity, reduce cognitive damage, and improve neuropsychic behavior in mice, making it a promising molecule to treat depression. Objectives: This study investigated the antidepressant-like action of MTDZ in mice and its potential mechanisms of action. Results: Molecular docking assays were performed and suggested a potential inhibition of monoamine oxidase A (MAO-A) by MTDZ. The toxicity study revealed that MTDZ displayed no signs of toxicity, changes in oxidative parameters, or alterations to biochemistry markers, even at a high dose of 300 mg/kg. In behavioral tests, MTDZ administration reduced immobility behavior during the forced swim test (FST) without adjusting the climbing parameter, suggesting it has an antidepressant effect. The antidepressant-like action of MTDZ was negated with the administration of 5-HT1A, 5-HT1A/1B, and 5-HT3 receptor antagonists, implying the involvement of serotonergic pathways. Moreover, the antidepressant-like action of MTDZ was linked to the NO system, as L-arginine pretreatment inhibited its activity. The ex vivo assays indicated that MTDZ normalized ATPase activity, potentially linking this behavior to its antidepressant-like action. MTDZ treatment restricted MAO-A activity in the cerebral cortices and hippocampi of mice, proposing a selective inhibition of MAO-A associated with the antidepressant-like effect of the compound. Conclusions: These findings suggest that MTDZ may serve as a promising antidepressant agent due to its selective inhibition of MAO-A and the involvement of serotonergic and NO pathways [ABSTRACT FROM AUTHOR]

Purpose: This study aims to evaluate whether the third-trimester soluble fms-like tyrosine kinase-1 (sFlt-1) serum levels could be related to placenta accreta spectrum (PAS) disorders and the severity of postpartum blood loss. Methods: This was a nested case–control study which compared serum sFlt-1 level between gravid women with or without PAS disorders. Spearman correlation analysis was conducted to explore the relationship between sFlt-1 level and the volume of postpartum blood loss. Confounding factors were adjusted to avoid the impact on the results. Results: Sixty gravid women were enrolled: 36 women in the PAS group and 24 women in the non-PAS group. Women in the PAS group had a median sFlt-1 level of 9407.1 [2745.9–21,691.5] pg/ml, whereas women in the non-PAS group had a median sFlt-1 level of 25,779.2 [14317.1–35,626.7] pg/ml, (p < 0.001). The sFlt-1 level was negatively related to the volume of postpartum blood loss (r = − 0.358, p = 0.041). After adjusting for maternal age and gestational age at blood taking, sFlt-1 level showed no significant relationship with PAS disorders (p = 0.245) and postpartum blood loss (p = 0.526). Conclusion: Third-trimester sFlt-1 serum level is not independently associated with PAS disorders or postpartum blood loss after adjusting for confounding factors. [ABSTRACT FROM AUTHOR]

Blueberries are vulnerable to chilling injury (CI). This can lead to limited longevity when they are subjected to cold storage conditions. This study investigated the effectiveness of a preharvest spray containing 0.02% hexanal in reducing CI and improving the postharvest storage quality of 'Star' and 'Biloxi' blueberries. The blueberries were stored for a period of 5 weeks at 2 °C and in 90% relative humidity (RH). The findings revealed that the preharvest hexanal spraying of both cultivars delayed senescence by mitigating CI, as evidenced by the bolstering of the antioxidant defense system through increased superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), catalase (CAT), and phenylalanine ammonia lyase (PAL) enzyme activity. The treated fruit also maintained elevated levels of total phenol content (TPC), total flavonoids (TFC), and vitamin C, demonstrating enhanced free radical scavenging capacity (FRSC), while exhibiting reduced polyphenoloxidase (PPO) activity, and reduced malondialdehyde (MDA), and H2O2 content in comparison with the control group. The preharvest hexanal treatment also suppressed fruit softening by maintaining greater firmness and higher membrane stability index (MSI) scores, inhibiting the activity of polygalacturonase (PG), pectinmethylesterase (PME), xylanase, and α‐amylase, and reducing microbial counts (MC) and incidence of decay (DI) in comparison with the control. Preharvest hexanal treatment also improved the overall storage quality by reducing weight loss, total soluble solids (TSS), pH, and the TSS/acid ratio, while increasing titratable acidity (TA) in comparison with the control during cold storage. The findings suggest that hexanal, as a preharvest application, delays senescence effectively and preserves overall quality by enhancing cold tolerance through antioxidant defense mechanisms in blueberry storage under cold conditions. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

This investigation employed different ANN infrastructures for predicting the quality of sugarcane juice under varying microfluidization pressures (50–200 MPa) and cycles (1–7) which was previously unexplored. Two hidden layer (HL) activation functions (tansigmoid, logsigmoid) and learning algorithms (LM, GDX) with varying hidden layer neurons (HLNs) were tested to predict the color, total phenol content, total flavonoid content, chlorophyll content, total and reducing sugars, polyphenol oxidase activity, peroxidase activity, sucrose neutral invertase activity, aerobic plate count, yeast and mold count, particle size, sensory score and sedimentation rate of sugarcane juice under different microfluidization processing conditions. Results showed that the combination of LM + logsigmoid, GDX + logsigmoid and GDX + tansigmoid produced > 90% prediction accuracy. Among these models, GDX + tansigmoid exhibited 91.7% accuracy on training, and 96% accuracy on testing using relatively lower number of neurons (10 HLNs), and was therefore selected to predict the quality characteristics of sugarcane juice. [ABSTRACT FROM AUTHOR]

Pectic enzymes are an eco-friendly alternative to improve extraction processes for the food industry. The objective of the research was to obtain pectinase from the pulp of soursop (Annona muricata L.) and cherimoya (Annona cherimola L.), as an alternative to reduce agro-industrial waste in Colombia; involving the study of the factors influencing the protein content of the enzymes extracted from different stages and types of fruits, the conditions for enzymatic activity, and the potential application in the food industry, particularly in extraction processes and juice clarification. A factorial design 23, spectrometric and potentiometric techniques evidenced that the protein content depends on factors such as fruit stage (suitable "s" and unsuitable "u" fruits for consumption) and fruit type (soursop and cherimoya). The higher protein content was obtained by the precipitation method in comparison to ion exchange chromatography. Cherimoya fruit presented a higher protein content of 326 ± 26 mg by precipitation, however, the soursop samples evidenced higher enzymatic activity, 2.2 ± 0.1 U/mL. The pectinase obtained was an acidic enzyme of type pectin methylesterase with an enzymatic activity of 27.0 ± 0.9 U/mL, under the optimal conditions of pH 4 and temperature of 40 °C. The application in blackberry juice evidenced a decrease in the absorbance and pH changes. From the current research, the pulp of soursop and cherimoya was proven as a cheap source to obtain pectinase with enzymatic activity for juice clarification, and this process is an alternative to reduce the negative impact on the environment and an economical source for the local producers. [ABSTRACT FROM AUTHOR]

Introduction: This study aimed to identify the pathogen responsible for leaf blight in Cyclocodon lancifolius, investigate its biological characteristics, and identify effective synthetic fungicides. Additionally, this study examined changes in physiological and biochemical indices of leaves following pathogen infection and screened biocontrol bacteria that inhibit the pathogen growth, providing a scientific basis for preventing and managing leaf blight in C. lancifolius. Methods: Pathogens were isolated from the interface of healthy and infected leaf tissues and identified through morphological and molecular biological methods. Amplification and sequencing of three genomic DNA regions--internal transcribed spacer region, translation elongation factor 1-a, and glyceraldehyde-3-phosphate dehydrogenase of ribosomal DNA--were performed, followed by the construction of a phylogenetic tree. The biological characteristics of pathogens under various temperature and pH conditions and different nitrogen and carbon sources were analyzed using the mycelial growth rate method. The antifungal effects of 13 chemical agents were evaluated using the poisoned medium method and mycelial growth rate method. Changes in physiological and biochemical indicators post-infection were also assessed. An antagonistic experiment was conducted to screen for biocontrol bacteria. Results: A total of 29 potential pathogenic strains were isolated from infected leaf tissues, with Koch's Postulates confirming Stemphylium lycopersici as a key pathogen causing the disease. Growth analysis of S. lycopersici revealed optimal growth at 20°C and pH 6, with lactose or maltose serving as the most suitable carbon source and histidine as the preferred nitrogen source. Among the 13 synthetic fungicides tested, strain DHY4 exhibited the greatest sensitivity to 400 g/L flusilazole. Significant differences (p < 0.05) were observed in superoxide dismutase, phenylalanine ammonia-lyase, peroxidase, polyphenol oxidase, catalase, and malondialdehyde levels between treated and control groups 3 days post-inoculation. The biocontrol strain DYHS2, identified as a strain of Bacillus subtilis, demonstrated an inhibition rate of 51.80% against S. lycopersici in dual-culture experiments and showed a relative inhibition rate of 78.82% in detached leaf assays. Discussion: These findings provide valuable insights into the newly identified causal agent of leaf blight in C. lancifolius and its biological characteristics, underscoring the potential of B. subtilis DYHS2 and synthetic fungicides such as flusilazole as effective disease management strategies. [ABSTRACT FROM AUTHOR]

Psilocybin, the natural hallucinogen from Psilocybe (magic) mushrooms, is a highly promising drug candidate for the treatment of depression and several other mental health conditions. Biosynthesis of psilocybin from the amino acid l‐tryptophan involves four strictly sequential modifications. The third of these, ATP‐dependent phosphorylation of the intermediate 4‐hydroxytryptamine, is catalysed by PsiK. Here we present a crystallographic analysis and a structure‐based mutagenesis study of this kinase, providing insight into its mode of substrate recognition. The results of our work will support future bioengineering efforts aimed at generating variants of psilocybin with enhanced therapeutic properties. [ABSTRACT FROM AUTHOR]

Tunnels in enzymes with buried active sites are key structural features allowing the entry of substrates and the release of products, thus contributing to the catalytic efficiency. Targeting the bottlenecks of protein tunnels is also a powerful protein engineering strategy. However, the identification of functional tunnels in multiple protein structures is a non-trivial task that can only be addressed computationally. We present a pipeline integrating automated structural analysis with an in-house machine-learning predictor for the annotation of protein pockets, followed by the calculation of the energetics of ligand transport via biochemically relevant tunnels. A thorough validation using eight distinct molecular systems revealed that CaverDock analysis of ligand un/binding is on par with time-consuming molecular dynamics simulations, but much faster. The optimized and validated pipeline was applied to annotate more than 17,000 cognate enzyme–ligand complexes. Analysis of ligand un/binding energetics indicates that the top priority tunnel has the most favourable energies in 75% of cases. Moreover, energy profiles of cognate ligands revealed that a simple geometry analysis can correctly identify tunnel bottlenecks only in 50% of cases. Our study provides essential information for the interpretation of results from tunnel calculation and energy profiling in mechanistic enzymology and protein engineering. We formulated several simple rules allowing identification of biochemically relevant tunnels based on the binding pockets, tunnel geometry, and ligand transport energy profiles. Scientific contributions The pipeline introduced in this work allows for the detailed analysis of a large set of protein–ligand complexes, focusing on transport pathways. We are introducing a novel predictor for determining the relevance of binding pockets for tunnel calculation. For the first time in the field, we present a high-throughput energetic analysis of ligand binding and unbinding, showing that approximate methods for these simulations can identify additional mutagenesis hotspots in enzymes compared to purely geometrical methods. The predictor is included in the supplementary material and can also be accessed at https://github.com/Faranehhad/Large-Scale-Pocket-Tunnel-Annotation.git. The tunnel data calculated in this study has been made publicly available as part of the ChannelsDB 2.0 database, accessible at https://channelsdb2.biodata.ceitec.cz/. [ABSTRACT FROM AUTHOR]

The catalytic therapy for abundant •OH, 1O2, and •O2− provides an efficient tactic and methodology for rapid tumor/bacteria killing, whereas the limitation is focused on the catalytic efficiency owing to the in‐built energy band of catalytic materials. Thus, d‐band center tuning BioHJzyme is devised, which is composed of zinc tellurate/manganese dioxide anchored by glucose oxidase (GOx) with anti‐tumor and anti‐bacteria properties for postoperative infected wound regeneration of tumor. The GOx depletes glucose to produce H2O2, intercepting the glucose metabolism. The D‐BioHJzyme can catalyze the produced H2O2 to highly lethal •OH with POD‐mimetic activity owing to the tunned d‐band center strategy decreasing the adsorption energy of the intermediate in the catalytical process, while the oxygen originated from H2O2 and Mn2+ can be catalyzed into 1O2. The GPx‐mimetic activity of it can impair the antioxidant system of tumor. In vivo studies show that the BioHJzyme exhibits robust abilities against bacteria and tumors by •OH/1O2 production and promotes the apoptosis owing the Te. Besides, the BioHJzyme can accelerate cutaneous regeneration by M1/M2 regulation and the angiogenesis/collagen deposition. This work enlightens a powerful platform for the remedy of postoperative infected wound regeneration of tumor resection using an engineered BioHJzyme. [ABSTRACT FROM AUTHOR]

Inflammatory bowel disease (IBD) manifests as inflammation in the colon, rectum, and ileum, presenting a global health concern with increasing prevalence. Therefore, effective anti-inflammatory therapy stands as a promising strategy for the prevention and management of IBD. However, conventional nano drug delivery systems (NDDSs) for IBD face many challenges in targeting the intestine, such as physiological and pathological barriers, genetic variants, disease severity, and nutritional status, which often result in nonspecific tissue distribution and uncontrolled drug release. To address these limitations, stimulus-responsive NDDSs have received considerable attention in recent years due to their advantages in providing controlled release and enhanced targeting. This review provides an overview of the pathophysiological mechanisms underlying IBD and summarizes recent advancements in microenvironmental stimulus-responsive nanocarriers for IBD therapy. These carriers utilize physicochemical stimuli such as pH, reactive oxygen species, enzymes, and redox substances to deliver drugs for IBD treatment. Additionally, pivotal challenges in the future development and clinical translation of stimulus-responsive NDDSs are emphasized. By offering insights into the development and optimization of stimulus-responsive drug delivery nanoplatforms, this review aims to facilitate their application in treating IBD. This review highlights recent advancements in stimulus-responsive nano drug delivery systems (NDDSs) for the treatment of inflammatory bowel disease (IBD). These innovative nanoplatforms respond to specific environmental triggers, such as pH reactive oxygen species, enzymes, and redox substances, to release drugs directly at the inflammation site. By summarizing the latest research, our work underscores the potential of these technologies to improve drug targeting and efficacy, offering new directions for IBD therapy. This review is significant as it provides a comprehensive overview for researchers and clinicians, facilitating the development of more effective treatments for IBD and other chronic inflammatory diseases. [Display omitted] [ABSTRACT FROM AUTHOR]

Semi‐artificial photosynthetic system (SAPS) that combines enzymes or cellular organisms with light‐absorbing semiconductors, has emerged as an attractive approach for nitrogen conversion, yet faces the challenge of reaction pathway regulation. Herein, we find that photoelectrons can transfer from the −C≡N groups at the edge of cyano‐rich carbon nitride (g‐C3N4‐CN) to nitrate reductase (NarGH), while the direct electron transfer to nitrite reductase (cd1NiR) is inhibited due to the physiological distance limit of active sites (>14 Å). By means of the directional electron transfer between g‐C3N4‐CN and extracted biological enzymes, the product of the denitrification reaction was switched from inert N2 to usable nitrite with an unprecedented selectivity of up to 95.3 %. The converted nitrite could be further utilized by anammox microbiota and dissimilatory nitrate reduction to ammonia (DNRA) microorganisms, doubling the efficiency of total nitrogen removal (96.5±2.3 %) for biological nitrogen removal and ammonia generation (12.6 mg NH4+‐N L−1 h−1), respectively. Thus, our work paves an appealing way for the sustainable treatment and utilization of nitrate for ammonia fuel production by strategically regulating the electron transfer pathway across the biotic‐abiotic interface. [ABSTRACT FROM AUTHOR]

Plastic pollution is one of the main worldwide environmental concerns. Our lifestyle involves persistent plastic consumption, aggravating the low efficiency of wastewater treatment plants in its removal. Nano/microplastics are accumulated in living beings, pushing to identify new water remediation strategies to avoid their harmful effects. Enzymes (e. g., Candida rugosa‐CrL) are known natural plastic degraders as catalysts in depolymerization reactions. However, their practical use is limited by their stability, recyclability, and economical concerns. Here, enzyme immobilization in metal‐organic frameworks (CrL_MOFs) is originally presented as a new plastic degradation approach to achieve a boosted plastic decomposition in aqueous systems while allowing the catalyst cyclability. Bis‐(hydroxyethyl)terephthalate (BHET) was selected as model substrate for decontamination experiments for being the main polyethylene terephthalate (PET) degradation product. Once in contaminated water, CrL_MOFs can eliminate BHET (37 %, 24 h), following two complementary mechanisms: enzymatic degradation (CrL action) and byproducts adsorption (MOF effect). As a proof‐of‐concept, the capacity of a selected CrL_MOF composite to eliminate the BHET degradation products and its reusability are also investigated. The potential of these systems is envisioned in terms of improving enzyme cyclability, reducing costs along with feasible co‐adsorption of plastic byproducts and other harmful contaminants, to successfully remove them in a single step. [ABSTRACT FROM AUTHOR]

Background: Malt is a vital ingredient in the brewing and food industries, as it influences the sweetness, flavor, and color of the final products. The quality and soluble solids content of barley-derived malt depends on temperature, enzyme type, and grinding roller interval. This study aimed to determine the optimal conditions for maximizing the soluble solids in Iranian malt. Methods: Malt samples were treated at 57 and 72 °C, employing glucan maltohydrolase (GMH) and glucohydrolase (GH) enzymes, and grinding roller intervals of 0.6, 0.8, and 1 mm. The malt samples were then subjected to chemical analyses, including Brix measurement, iodine test, and pH determination. Results: The results showed that higher temperatures increased the Brix value and decreased the pH value of the malt samples. No significant differences in Brix and pH were observed between the GMH and GH enzymes. Smaller grinding roller intervals (0.6 mm) increased the Brix value and decreased the pH. The optimal conditions for maximizing the soluble solids content were 72 °C with the GMH enzyme and a 0.6 mm roller interval (Brix = 7.8 g/100 cc, pH = 5.32). The temperature significantly influenced the iodine test, confirming starch hydrolysis into sugar at 72 °C. Conclusion: This study recommends reducing the grinding roller interval from 0.8 to 0.6 mm and increasing the temperature from 57 to 72 °C to significantly enhance the malting process. Therefore, using a 72 °C temperature and 0.6 mm roller gap is advised for better malting efficiency. [ABSTRACT FROM AUTHOR]

The formation of biofilm in different places and the failure to effectively remove it by the usual disinfection methods is due to its structure and the rich genetic resource available in it to deal with disinfectants. These impenetrable structures and diverse microbial genetics have caused biofilm pollution in different industries like the food industry, the medicine industry, the hospitals and the water distribution system, resulting in pathogenicity and reduction of industrial quality. An efficient way to deal with the resistant population of biofilm‐forming microbes is the use of hurdle technology including enzymes and essential oils. Enzymes reduce the resistance of the biofilm structure due to degradation of its extracellular polymer matrix (EPS) by their abilities to break down the organic molecules, and then the essential oils weaken the cells by penetrating the lipid membrane of the cell and destroying its integrity; as a result, the biofilm will be destroyed. The advantage of this hurdle technology is the environmental friendly of both methods, which reduces concerns about the use of chemical disinfection methods, but on the other hand, due to the sensitivity of enzymes as biological agents also the expensiveness of this technique and the considerations of working with essential oils as volatile and unstable liquids should abandon the routine methods of applying this disinfectant to biofilm and go for the microencapsulation method, which as a protective system increases the effectiveness of enzymes and essential oils as antibiofilm agents. [ABSTRACT FROM AUTHOR]

Crop rotation could effectively alleviate continuous cropping problems, but how it affects soil organic carbon (SOC) components and chemical structures in case of different duration of continuous cropping (CC) remains unclear. In this study, we selected cotton fields that had been continuously planted for 5, 10, 15, and 20 years for separate soybean rotations (SR). The mechanism of SR to improve soil quality in different duration of CC was determined by SOC components and their chemical structure. The results showed that the content of soil available nutrients, SOC, microbial biomass carbon and soluble organic carbon in the SR treatments significantly increased compared with those in the CC treatment (p < 0.05), but particulate organic carbon content decreased by 24.89–81.10%. Compared with CC in different years, SR treatments significantly reduced soil humus content. SR in 5, 10 and 15 years of CC cotton fields significantly increased the content of sucrase and decreased that of polyphenol oxidase. Compared with CC fields in different years (5-y, 10-y, 15-y, 20-y), SR increased the relative peak intensity of polysaccharides and decreased the relative peak intensity of alcohols and phenols. Among them, the relative peak intensity of R5-y, R10-y and R20-y aliphatic compounds increased, while that of R5-y, R15-y and R20-y aromatic compounds decreased, and R10-y recalcitrant components (Aliphatic + Aromatic) increased. [ABSTRACT FROM AUTHOR]

The identification of novel acetylcholinesterase inhibitors holds significant relevance in the treatment of Alzheimer's disease (AD), the prevailing form of dementia. The exploration of alternative inhibitors to the conventional acetylcholinesterase inhibitors is steadily gaining prominence. Quinones, categorized as plant metabolites, represent a specific class of compounds. In this study, the inhibitory effects of various naphthoquinone derivatives, along with anthraquinone and its derivatives, on the acetylcholinesterase (AChE) enzyme were investigated for this purpose. An in vitro investigation was conducted to examine the effects of these compounds in order to clarify the possible mechanism of inhibition in the interaction between the enzyme and chemicals. In addition, an in silico investigation was carried out to understand the conceivable inhibitor binding process to the enzyme's active site. The acquired outcomes corroborated the in vitro results. The AChE enzyme was found to be effectively inhibited by both naphthoquinones and anthraquinones, with inhibition constant (KI) values ranging from 0.014 to 0.123 μM (micormolar). The AChE enzyme was inhibited differently by this quinone and its derivatives. Although derivatives of naphthoquinone and anthraquinone exhibited a competitive inhibitory effect, derivatives of anthraquinone exhibited a noncompetitive inhibition effect. Furthermore, because it had the lowest KI value of any of these substances, 1,5‐dihydroxyanthraquinone (1c) was shown to be the most potent inhibitor. The findings will add to the body of knowledge on the creation of fresh, potent, and successful treatment approaches. [ABSTRACT FROM AUTHOR]

Extracellular enzymes produced by fungi can serve as a screening tool for entomopathogen candidates. Enzyme secretion ability is important in determining the virulence level of an entomopathogenic fungus. This study aims to see the potential of fungal isolates through chitinolytic, lipolytic, and proteolytic activities of five entomopathogenic fungal isolated from Bactrocera sp. imago from citrus plants. The research was carried out at the Microbiology Laboratory of the Department of Biology at the University of Lampung. Enzymatic activity is known through the method of observing the clear zone formed on agar media. Quantitative tests based on the calculation of enzymatic index values showed that Nigrospora sp. (IB1) had lipolytic activity, Penicillium sp. 2 (IB5) isolates had chitinolytic, lipolytic, and proteolytic activities. The isolates of Penicillium sp. 1 (IB2), Phytopthora sp. (IB3), and Gliocladium sp. (IB4) did not show the activity of the three enzymes. [ABSTRACT FROM AUTHOR]

Bacillus circulans xylanase (BcX) from the glycoside hydrolase family 11 degrades xylan through a retaining, double‐displacement mechanism. The enzyme is thought to hydrolyze glycosidic bonds in a processive manner and has a large, active site cleft, with six subsites allowing the binding of six xylose units. Such an active site architecture suggests that oligomeric xylose substrates can bind in multiple ways. In the crystal structure of the catalytically inactive variant BcX E78Q, the substrate xylotriose is observed in the active site, as well as bound to the known secondary binding site and a third site on the protein surface. Nuclear magnetic resonance (NMR) titrations with xylose oligomers of different lengths yield nonlinear chemical shift trajectories for active site nuclei resonances, indicative of multiple binding orientations for these substrates for which binding and dissociation are in fast exchange on the NMR timescale, exchanging on the micro‐ to millisecond timescale. Active site binding can be modeled with a 2 : 1 model with dissociation constants in the low and high millimolar range. Extensive mutagenesis of active site residues indicates that tight binding occurs in the glycon binding site and is stabilized by Trp9 and the thumb region. Mutations F125A and W71A lead to large structural rearrangements. Binding at the glycon site is sensed throughout the active site, whereas the weak binding mostly affects the aglycon site. The interactions with the two active site locations are largely independent of each other and of binding at the secondary binding site. [ABSTRACT FROM AUTHOR]

Introduction: Bone is constantly remodeling to maintain the structure of the bone tissue that composes it. This is susceptible to changes that can be both favorable and harmful, marked by certain variables such as age, sex, diseases, hormonal alterations, trauma, among others. Method: Due to the above, the idea was raised to study which treatment is better in the regeneration of bone tissue, comparing pharmacological treatments (cathepsin K inhibitors) against micro-vibratory stimulation or non-pharmacological treatment. Objective: To carry out a systematic review of treatments with micro-vibrations and cathepsin K inhibitors in relation to bone remodeling. To make a comparison between the effectiveness of treatment based on micro-vibrations and with cathepsin K inhibitors, a systematic review was carried out in nine databases (Wiley Online Library, Pubmed, Google Academic, Scopus, Science Direct, Scielo, Medline, EBSCO and SpringerLink). Results: Twenty articles were included in this study, which demonstrated that both treatments improve the bone remodeling process. Conclusions: Taking into consideration the systematic review carried out, it has been determined that the treatment of low-intensity, high-frequency micro-vibrations increases the outer cortex; however, the use of cathepsin k inhibitors promises innovative treatments in tissue regeneration. [ABSTRACT FROM AUTHOR]

Enzyme catalysis in reactors for industrial applications usually require an external intervention of the species involved in the chemical reactions. We analyze the most elementary open enzyme catalysis with competitive inhibition where a time-dependent inflow of substrate and inhibitor supplies is modeled by almost periodic functions. We prove global stability of an almost periodic solution for the non-autonomous dynamical system arising from the mass-law action. This predicts a well behaved situation in which the reactor oscillates with global stability. This is a first case study in the path toward broader global stability results regarding intraspecific and monotone open reaction networks. [ABSTRACT FROM AUTHOR]

[Display omitted] Here, we report the preparation of a novel Janus nanoparticle with opposite Ir and mesoporous silica nanoparticles through a partial surface masking with toposelective modification method. This nanomaterial was employed to construct an enzyme-powered nanomachine with self-propulsion properties for on-command delivery. The cargo-loaded nanoparticle was provided with a pH-sensitive gate and unit control at the mesoporous face by first attaching boronic acid residues and further immobilization of glucose oxidase through reversible boronic acid esters with the carbohydrate residues of the glycoenzyme. Addition of glucose leads to the enzymatic production of H 2 O 2 and gluconic acid, being the first compound catalytically decomposed at the Ir nanoparticle face producing O 2 and causing the nanomachine propulsion. Gluconic acid leads to a pH reduction at the nanomachine microenvironment causing the disruption of the gating mechanism with the subsequent cargo release. This work demonstrates that enzyme-mediated self-propulsion improved release efficiency being this nanomotor successfully employed for the smart release of Doxorubicin in HeLa cancer cells. [ABSTRACT FROM AUTHOR]

BACKGROUND AND OBJECTIVES: Trichoderma species (Trichoderma sp.) is commonly found in nature, particularly in soil and roots, often thriving in plants rich in cellulose such as oil palm. Therefore, this study aimed to characterize Trichoderma sp. in each substrate as well as to assess cellulolytic and ligninolytic potential in vitro using a specific medium. Characterization and utilization of the plant were examined to efficiently manage empty fruit bunch (EFB) waste in Indonesian oil palm plantations. By exploring the biological, ecological, and biochemical attributes of Trichoderma sp., this study examined the mechanisms associated with efficacy in bioconversion processes and waste remediation. METHODS: A survey method was used to determine sampling locations, and purposive sampling was utilized in the field. Furthermore, several stages were undertaken including isolation, identification, as well as testing of morphology and potential in vitro. FINDINGS: The results showed that the growth characteristics of Trichoderma sp. in filling a petri dish spanned approximately five days, forming colonies with a dark green hue. Microscopically, the isolates consisted of conidiophores, phialides, and conidia. Meanwhile, in vitro potential was assessed through cellulolytic and lignocellulolytic assays, with each isolate demonstrating varying capabilities in breaking down cellulose and lignocellulose. The highest cellulase activity (3.55 units per gram) was observed in enzyme starter granules treated with air drying. The highest viability per gram was found in spore inoculum (17.0 x 107) with air drying treatment. The greatest reduction in weight of oil palm EFB waste (65.3 percent) was achieved through the application of Trichoderma sp. enzyme starter granules with air drying treatment. Furthermore, eight isolates demonstrated cellulolytic and ligninolytic potential. Isolates sourced from soil had rapid growth on starter and baglog mediums, indicating potential for subsequent field applications as a biodegradation agent for organic waste. CONCLUSION: Based on the macroscopic (shape and color of mycelium) and microscopic characteristics (mycelium form), a total of eight Trichoderma sp. isolates were obtained. During the potential test, it was observed each isolate had different cellulolytic and ligninolytic activities. This study provided valuable insights into the diversity and enzymatic potential of Trichoderma sp. fungi, underscoring the importance of selecting suitable strains and optimizing cultivation conditions for various biotechnological applications. By exploring cellulolytic and ligninolytic capabilities, the results contributed to the development of more efficient and sustainable strategies for organic waste management and bioconversion. In general, this study represented a significant milestone in the quest for sustainable waste management solutions in agriculture, offering a promising pathway towards mitigating environmental impact and promoting the long-term sustainability of oil palm cultivation. [ABSTRACT FROM AUTHOR]

This comprehensive review explores the diverse landscape of plant protein resources, focusing on novel extraction and precipitation methods that contribute to the advancement of sustainable and efficient protein production. The increasing global demand for plant‐based proteins as a key component of balanced diets has spurred research into optimizing extraction processes from various plant sources. The review encompasses a wide range of plant protein resources, including legumes, cereals, oilseeds, and other plant‐based alternatives. Special emphasis is given to innovative techniques in protein extraction, such as enzyme‐assisted extraction, aqueous extraction, and emerging technologies like ultrasound and microwave‐assisted methods. Additionally, the review investigates novel precipitation methods employed to isolate and purify plant proteins, shedding light on techniques like isoelectric precipitation, salting‐out, and membrane filtration. The integration of sustainable practices in protein extraction, coupled with a focus on minimizing environmental impact, is a central theme throughout the review. By synthesizing current research findings, this review aims to provide a comprehensive overview of the state‐of‐the‐art in plant protein extraction and precipitation methods, offering valuable insights for researchers, industry professionals, and policymakers involved in the rapidly evolving field of plant‐based protein production. [ABSTRACT FROM AUTHOR]

Using free microorganisms for industrial processes has some limitations, such as the extensive consumption of substrates for growth, significant sensitivity to the microenvironment, and the necessity of separation from the product and, therefore, the cyclic process. It is widely acknowledged that confining or immobilizing cells in a matrix or support structure enhances enzyme stability, facilitates recycling, enhances rheological resilience, lowers bioprocess costs, and serves as a fundamental prerequisite for large-scale applications. This report summarizes the various cell immobilization methods, including several synthetic (polyvinylalcohol, polyethylenimine, polyacrylates, and Eudragit) and natural (gelatin, chitosan, alginate, cellulose, agar–agar, carboxymethylcellulose, and other polysaccharides) polymeric materials in the form of thin films, hydrogels, and cryogels. Advancements in the production of well-known antibiotics like penicillin and cephalosporin by various strains were discussed. Additionally, we highlighted cutting-edge research related to strain producers of peptide-based antibiotics (polymyxin B, Subtilin, Tyrothricin, varigomycin, gramicidin S, friulimicin, and bacteriocin), glusoseamines, and polyene derivatives. Crosslinking agents, especially covalent linkers, significantly affect the activity and stability of biocatalysts (penicillin G acylase, penicillinase, deacetoxycephalosporinase, L-asparaginase, β-glucosidase, Xylanase, and urease). The molecular weight of polymers is an important parameter influencing oxygen and nutrient diffusion, the kinetics of hydrogel formation, rigidity, rheology, elastic moduli, and other mechanical properties crucial for long-term utilization. A comparison of stability and enzymatic activity between immobilized enzymes and their free native counterparts was explored. The discussion was not limited to recent advancements in the biopharmaceutical field, such as microorganism or enzyme immobilization, but also extended to methods used in sensor and biosensor applications. In this study, we present data on the advantages of cell and enzyme immobilization over microorganism (bacteria and fungi) suspension states to produce various bioproducts and metabolites—such as antibiotics, enzymes, and precursors—and determine the efficiency of immobilization processes and the optimal conditions and process parameters to maximize the yield of the target products. [ABSTRACT FROM AUTHOR]

Anthropogenic activities have led to hydrocarbon spills, and while traditional bioremediation methods are costly and time-consuming, recent research has focused on engineered enzymes for managing pollutant. The potential of enzymes for resolving wax flow problems in the petroleum industry remains unexplored. This paper offers a comprehensive review of the current state of research activities related to the bioremediation of petroleum-polluted sites and the biodegradation of specific petroleum hydrocarbons. The assayed enzymes that took part in the degradation were discussed in detail. Lipase, laccase, alkane hydroxylase, alcohol dehydrogenase, esterase, AlkB homologs and cytochrome P450 monooxygenase are among the enzymes responsible for the degradation of more than 50% of the hydrocarbons in contaminated soil and wastewater and found to be active on carbon C8 to C40. The possible biodegradation mechanism of petroleum hydrocarbons was also elucidated. The enzymes' primary metabolic pathways include terminal, subterminal, and ω-oxidation. Next, given the successful evidence of the hydrocarbon treatment efficiency, the authors analyzed the opportunity for the enzymatic degradation approach if it were to be applied to a different scenario: managing wax deposition in petroleum-production lines. With properties such as high transformation efficiency and high specificity, enzymes can be utilized for the treatment of viscous heavy oil for transportability, evidenced by the 20 to 99% removal of hydrocarbons. The challenges associated with the new approach are also discussed. The production cost of enzymes, the characteristics of hydrocarbons and the operating conditions of the production line may affect the biocatalysis reaction to some extent. However, the challenges can be overcome by the usage of extremophilic enzymes. The combination of technological advancement and deployment strategies such as the immobilization of a consortium of highly thermophilic and halotolerant enzymes is suggested. Recovering and reusing enzymes offers an excellent strategy to improve the economics of the technology. This paper provides insights into the opportunity for the enzymatic degradation approach to be expanded for wax deposition problems in pipelines. [ABSTRACT FROM AUTHOR]

The palm oil industry has contributed substantially to environmental pollution due to the large volume of oil extraction by-products known as palm oil mill effluents (POME). Compared to several industrial effluents, POME is about 100 times more contaminated in terms of biochemical and chemical oxygen demand. Aerobic granular sludge technology is considered a more effective method for treating POME than traditional activated sludge systems. Nonetheless, the lengthy start-up phase of the granulation process is a significant disadvantage of this method. In this study, eco-eco-enzymes, a multifunctional liquid with high enzymatic activity, was tested to potentially accelerate the initial growth of granules. The study utilized a 5-set of 1-L bioreactors operated simultaneously, and samples were taken every 5 h for granulation and removal performance analysis. The results showed that the Spinach eco-enzyme had the highest potential to enhance initial granulation, with the highest surface hydrophobicity (52.6%), aggregation (80.7%), and settling performance (54.03 mL/g SS SVI) compared to banana, orange, and mango. Additionally, the Spinach eco-enzyme was the most efficient in removing total phosphorus, total ammonia nitrogen, and chemical oxygen demand (COD), with an 80%, 91.4%, and 70.4% respectively. These findings suggest that eco-enzymes, particularly the Spinach eco-enzyme, have the potential to act as a secondary treatment for treating POME with excellent initial granulation performance compared to banana, orange and mango eco-enzyme. [ABSTRACT FROM AUTHOR]

Pretreatment is compulsory to obtain the greatest possible biofuel production from lignocellulosic biomass. Among the many different pretreatment methods, enzymatic and ultrasonic pretreatments are attracting attention as environmentally friendly methods. In this framework, this study aims to optimize enzymatic and ultrasonic pretreatment to achieve high methane production by anaerobic digestion of lignocellulosic biomass consisting of agricultural residues. The optimization studies selected independent variables such as cellulase and β-glycosidase doses for enzymatic pretreatment and ultrasound power, reaction time, and solid content for ultrasonic pretreatment. The optimum conditions for enzymatic pretreatment were determined as 32.68FPU/gTS cellulase and 14.56 IU/gTS β-glucosidase concentrations to achieve a 60.15% increase in methane production. On the other hand, an 80.04% increase in methane production was obtained in the ultrasonic pretreatment under optimum conditions determined as 20 W ultrasound power, 60-min reaction time, and 5.46% solids content. The decrease in methane production by increasing the cellulose concentration in enzymatic pretreatment and by increasing the ultrasound power in ultrasonic pretreatment shows the importance of process optimization to achieve methane production. The theoretical energy productions were calculated and then compared with other pretreatment methods and the theoretical direct combustion energy. Since 90% energy recovery with ultrasonic pretreatment compared to direct combustion of lignocellulosic biomass was achieved, it is recommended to determine the effects of ultrasound in the pretreatment of other lignocellulosic biomass types. Thus, it will be possible to obtain energy from lignocellulosic waste in a much more environmentally friendly way compared to direct combustion. [ABSTRACT FROM AUTHOR]

In light of the growing demand for novel biocatalysts and enzyme production methods, this study aimed to evaluate the potential of Aspergillus tubingensis for producing lipase under submerged culture investigating the influence of culture time and inducer treatment. Moreover, this study also investigated conditions for the immobilization of A. tubingensis lipase by physical adsorption on styrene–divinylbenzene beads (Diaion HP-20), for these conditions to be applied to an alternative immobilization system with a packed-bed reactor. Furthermore, A. tubingensis lipase and its immobilized derivative were characterized in terms of their optimal ranges of pH and temperature. A. tubingensis was shown to be a good producer of lipase, obviating the need for inducer addition. The enzyme extract had a hydrolytic activity of 23 U mL−1 and achieved better performance in the pH range of 7.5 to 9.0 and in the temperature range of 20 to 50 °C. The proposed immobilization system was effective, yielding an immobilized derivative with enhanced hydrolytic activity (35 U g−1), optimum activity over a broader pH range (5.6 to 8.4), and increased tolerance to high temperatures (40 to 60 ℃). This research represents a first step toward lipase production from A. tubingensis under a submerged culture and the development of an alternative immobilization system with a packed-bed reactor. The proposed system holds promise for saving time and resources in future industrial applications. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the effects of three feed forms (pellet, crumble and mash) with or without CIBENZA enzyme supplementing (0.005%) on growth performance, blood parameters, and carcass traits of broiler. A total number of 576 unsexed one-day-old Cobb broiler chicks were grown over a 35-d period. Chicks were similar concerning body weight and sex. The experimental design was factorial 3x2; three forms of feed (pellet, crumble, and mash) with or without supplementing CIBENZA enzyme (0.005%) in diets. All diets were nearly iso-nitrogenous and iso-caloric based on digestible energy and contained similar levels of microelements. The results showed that chicks fed pellets diet recorded the highest final body weight, while those fed mash diet had the lowest value (P<0.001). Enzyme supplementing in diets led to a significant increase in final body weight by 1.04%, as compared with un-supplemented diets. Chicks fed pellets diet had the highest carcass weight percentage; while those fed mash diet had the lowest value (P<0.05). Serum total protein was significantly decreased with chicks fed mash diet, as compared with those fed pellets and crumble diets. Conclusively, it could be concluded that pellets feed form in broiler nutrition improved growth performance, feed efficiency, and blood metabolite profile when compared to crumble and mash feed form. Supplementation of protease enzyme in diets enhanced growth performance of broiler chickens. [ABSTRACT FROM AUTHOR]

The increasing global consumption of poultry meat has led to the generation of a vast quantity of feather keratin waste daily, posing significant environmental challenges due to improper disposal methods. A growing focus is on utilizing keratinous polymeric waste, amounting to millions of tons annually. Keratins are biochemically rigid, fibrous, recalcitrant, physiologically insoluble, and resistant to most common proteolytic enzymes. Microbial biodegradation of feather keratin provides a viable solution for augmenting feather waste's nutritional value while mitigating environmental contamination. This approach offers an alternative to traditional physical and chemical treatments. This review focuses on the recent findings and work trends in the field of keratin degradation by microorganisms (bacteria, actinomycetes, and fungi) via keratinolytic and proteolytic enzymes, as well as the limitations and challenges encountered due to the low thermal stability of keratinase, and degradation in the complex environmental conditions. Therefore, recent biotechnological interventions such as designing novel keratinase with high keratinolytic activity, thermostability, and binding affinity have been elaborated here. Enhancing protein structural rigidity through critical engineering approaches, such as rational design, has shown promise in improving the thermal stability of proteins. Concurrently, metagenomic annotation offers insights into the genetic foundations of keratin breakdown, primarily predicting metabolic potential and identifying probable keratinases. This may extend the understanding of microbial keratinolytic mechanisms in a complex community, recognizing the significance of synergistic interactions, which could be further utilized in optimizing industrial keratin degradation processes. [ABSTRACT FROM AUTHOR]

Backgrounds: Various physiological functions and reaction cascades, as well as disease progression in the living systems, are controlled by the activity of specific proteolytic enzymes. We conducted the study to evaluate protease activity by assessing peptide fragments from either conserved or labeled red blood cells (RBCs) with aminofluorescein (AF) in the reaction media. Methods: RBCs were incubated in media containing trypsin. Subsequently, the concentration of peptide fragments in the reaction media, resulted by the digestion with trypsin from conserved cells, was estimated by 3‐(4‐carboxybenzoyl)quinoline‐2‐carboxaldehyde (CBQCA) as an amine‐reactive fluorogenic reagent. In a second approach, we conjugated AF to the conserved RBCs and then exposed AF‐labeled RBCs to trypsin. This was followed by directly measuring the fluorescence intensity (FI) in the reaction media to estimate the concentration of AF‐labeled peptide fragments resulting from the enzyme's activity. Results: Show a concentration‐ and time‐dependent increase in FIs, reflecting the activity of trypsin as a proteolytic enzyme. The FIs increased significantly by 4 to 5 folds in samples treated with different enzyme concentrations, and by over 11 folds after 2 h incubation in media containing a 50 μL trypsin, as evidenced by CBQCA assays. Conclusion: These fast and affordable approaches could be applied with high reliability for the general estimation of protease activity in samples and customized for diagnostic purposes and prognostic evaluation in various diseases. [ABSTRACT FROM AUTHOR]

Human 5′‐3′ exonuclease PLD3, a member of the phospholipase D family of enzymes, has been validated as a therapeutic target for treating Alzheimer's disease. Here, we have determined the crystal structure of the luminal domain of the enzyme at 2.3 Å resolution, revealing a bilobal structure with a catalytic site located between the lobes. We then compared the structure with published crystal structures of other human PLD family members which revealed that a number of catalytic and lipid recognition residues, previously shown to be key for phospholipase activity, are not conserved or, are absent. This led us to test whether the enzyme is actually a phospholipase. We could not measure any phospholipase activity but the enzyme shows robust nuclease activity. Finally, we have mapped key single nucleotide polymorphisms onto the structure which reveals plausible reasons as to why they have an impact on Alzheimer's disease. [ABSTRACT FROM AUTHOR]

Glycolysis is a highly conserved biochemical process that involves the sequential conversion of glucose to pyruvate and the generation of adenosine triphosphate (ATP), generating energy for cellular functions. This is carried out by several enzymes that play an important role in glucose metabolism. The sulfonamide group of drugs is commonly used as an antibiotic that inhibits the bacterial folic acid production pathway, although its effects on glycolytic enzymes are unclear. Since altered glycolytic enzyme expression is linked to a variety of metabolic disorders as well as cancer, the quest for new ligands against glycolytic enzymes is never over. We revealed that different sulfonamide derivatives interact with glycolytic enzymes with varying binding affinities. Among the selected sulfonamides, sulfisoxazole and sulfamethazine had greater binding scores to certain glycolytic enzymes than others. In silico point mutation analysis predicts that sulfisoxazole and sulfamethazine interact with certain glycolytic enzymes in a residue‐independent manner. This research generates a repository of the 2D interactions of sulfonamide derivatives with glycolytic enzymes, binding energies, impacts of induced point mutations on the local environment, and changes in protein structural flexibility. In the future, sulfisoxazole and sulfamethazine may be repurposed as specific glycolytic enzyme inhibitors to combat a variety of glycolysis‐related metabolic disorders. [ABSTRACT FROM AUTHOR]

tRNAs have recently gained attention for their novel regulatory roles in translation and for their diverse functions beyond translation. One of the most remarkable aspects of tRNA biogenesis is the incorporation of various chemical modifications, ranging from simple base or ribose methylation to more complex hypermodifications such as formation of queuosine and wybutosine. Some tRNAs are transcribed as intron-containing pre-tRNAs. While the majority of these modifications occur independently of introns, some are catalyzed in an intron-inhibitory manner, and in certain cases, they occur in an intron-dependent manner. This review focuses on pre-tRNA modification, including introncontaining pre-tRNA, in both intron-inhibitory and intron-dependent fashions. Any perturbations in the modification and processing of tRNAs may lead to a range of diseases and disorders, highlighting the importance of understanding these mechanisms in molecular biology and medicine. [ABSTRACT FROM AUTHOR]

Seaweed industries are producing solid waste composting of about 60–70% w/w fiber, which is rich in cellulose. Conversion of cellulose-rich waste into sugar and ethanol can provide a zero-waste industry. The objectives of this study were to characterize the solid waste biomass (SWBS) of agar processing industries and convert it into sugar and ethanol. The chemical composition and morphology of SWBS samples from three agar extraction plants (SWBS EP1, SWBS EP2, and SWBS EP3) were examined. Results showed that the SWBS had 3.01–16.4% cellulose, hemicellulose of 3.41–11.53%, lignin of 5.49–6.45%, agar of 7.28–17.03%, and ash content of 59.18–78.75%. X-ray diffraction (XRD) analysis supported the Fourier transform infrared (FTIR) result while revealing that the filter aid of the SWBS EP3 sample was not the same with the two others. SWBS EP3 sample contained more biomass and less filter aid, and it was selected to convert into sugar and ethanol in a 250-L tank. The concentration biosugar reached 21.45 g/L with the addition of the cellulase after the agarase (A24S24), and 15 g/L when the cellulase was mixed together with the agarase (AS24). The highest ethanol production was obtained when agarase and cellulase were added simultaneously for 24-h saccharification followed by yeast fermentation. [ABSTRACT FROM AUTHOR]

On-site quick detection of acetylcholinesterase (AChE) activity is of great significance for diagnosing nervous diseases, screening drugs, and monitoring poisoning. Herein, we developed a system for sensitively regulating gold nanobipyramid (AuNBP) morphology on a paper-based analytical device (PAD) via an Au+-disproportionation system, and further developed a high-resolution paper-based multicolor sensor (PMS) for both on-site visual detection and laboratory precision quantification of AChE activity using the system as a signal reporter and a dual-enzyme strategy as a receptor. The paper-based Au+ disproportionation-mediated AuNBP morphology system not only has excellent stability, reproducibility, and robust resistibility against reducing substances existing in biological samples but also can effectively suppress the color interference from reagents and greatly improve the sensitivity of regulating AuNBP morphology transformation, making it capable of generating long-term stable and more colorful/clearer color changes corresponding to Au+ concentrations. The developed PMS exhibited eight color changes corresponding to AChE activity within 0.0–1.0 U/L and the color changes could be stably maintained for 30 days, which makes both on-site visual detection and laboratory sensitive quantification possible. The PMS was successfully used to detect AChE activity in serum and erythrocyte samples with a visual detection limit (LOD) of 50 mU/L, a spectrometry LOD of 26 mU/L, and a recovery of 81–102%. The results obtained with the PMS were consistent with those obtained with Ellman's method, verifying its reliability. All of the above features make the PMS a promising assay for the rapid detection of AChE activity in clinical diagnosis. Importantly, the proposed paper-based Au+ disproportionation-mediated AuNBP morphology system can serve as a promising general signal reporter for developing various sensitive multicolor colorimetric sensors for target detection in various fields. [ABSTRACT FROM AUTHOR]

Tetrahydrofolate and its derivatives participate in one-carbon transfer reactions in all organisms. The cellular form of tetrahydrofolate (THF) is modified by multiple glutamate residues and polyglutamylation plays a key role in organellar and cellular folate homeostasis. In addition, polyglutamylation of THF is known to increase the binding affinity to enzymes in the folate cycle, many of which can utilize polyglutamylated THF as a substrate. Here, we use Xray crystallography to provide a high-resolution view of interactions between the enzyme serine hydroxymethyltransferase (SHMT), which provides one carbon precursors for the folate cycle, and a polyglutamylated form of THF. Our 1.7 Å crystal structure of soybean SHMT8 in complex with diglutamylated 5-formyl- THF reveals, for the first time, a structural rearrangement of a loop at the entrance to the folate binding site accompanied by the formation of novel specific interactions between the enzyme and the diglutamyl tail of the ligand. Biochemical assays show that additional glutamate moieties on the folate ligand increase both enzyme stability and binding affinity. Together these studies provide new information on SHMT structure and function and inform the design of anti-folate agents. [ABSTRACT FROM AUTHOR]

This study evaluated the efficacy of a multi‐protease on the performance, egg quality and digestibility of laying hen. The study had four treatments: Control (without multi‐protease, CON), Pro 1.2 (CON + 0.1% multi‐protease), Pro 2.4 (CON + 0.2% multi‐protease) and Pro 3.6 (CON + 0.3% multi‐protease). Each treatment was replicated six times (replicate = experimental unit = one pen with 15 hens) to give a total of 360 layer hens of the Hy‐line breed. The study lasted for a total of 3 months (14 day adaptation period + 84 days experimental period). The effects of the additive were assessed on: the performance variables, egg quality and ileal amino acid (AA) digestibility. At the end of the study, dietary supplementation with Pro 2.4 and Pro 3.6 improved (p < 0.05) hen‐day egg production, egg mass and eggshell thickness compared with CON at the peak phase. Further improvements (p < 0.05) were observed in the digestibility of crude protein and AAs such as isoleucine, lysine, threonine and cysteine at Pro 2.4 and Pro 3.6 protease supplementation levels compared with CON, while arginine and alanine were higher (p < 0.05) at Pro 3.6 compared with CON. No differences were reported for other performances such as body weight, average daily feed intake, average egg weight, feed conversion ratio, eggshell hardness and all the egg qualities measured. Overall, the results from this study showed better efficacy at Pro 2.4 and Pro 3.6 on the performance of laying hen during the peak phase. [ABSTRACT FROM AUTHOR]