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Polyethylene (PE), widely utilized in everyday life, is notorious for its protracted degradation period, extending over decades, presenting an environmental hazard. Recently, there has been growing interest in utilizing microorganisms to aid in PE decomposition. Molecular docking and molecular dynamics simulations are valuable tools for understanding specific mechanisms and conducting initial screenings to support experimental research in this context. In this study, various enzymes, including lignin peroxidase, laccase, manganese peroxidase, and cutinase, sourced from Phanerodontia chrysosporium, Melanocarpus albomyces, and Fusarium vanettenii, were investigated. The docking simulations revealed that lignin peroxidase exhibited the most substantial binding interaction with PE, displaying a binding energy of - 4.69162 kcal mol -1 and an RMSD value of 0.93428 Å. Following lignin peroxidase in binding strength were laccase, manganese peroxidase, and cutinase. Furthermore, molecular dynamics simulations provided insights into the binding mechanisms. These simulations demonstrated stability over a 200-ns period, as indicated by RMSD and RMSF values below 0.2 nm. Additionally, the study delved into the interaction mechanisms between microorganisms and plastic molecules, enriching our understanding of this process. While the findings of this study may be considered modest, they contribute to a broader perspective and have the potential to influence more profound and significant research in the field., Competing Interests: Declarations Ethical approval This is an observational study. The Van Lang University Research Ethics Committee has confirmed that no ethical approval is required. Consent to participate This research was conducted without the involvement of human subjects. Consent for publication The authors have provided explicit consent for the publication of the research findings and associated materials, in accordance with ethical standards. It is important to note that the data and images referenced in this paper are unrelated to human subjects. Competing interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Bread industry generates significant amounts of by-products which are discarded every day with relevant economic and environmental repercussions, despite they still contain high concentrations of potentially exploitable nutrients. Aiming to develop new sustainable solutions, this study explored the synergistic application of enzymatic treatment and sourdough fermentation to re-cycle industrial bread by-products for new sourdough bread making. Lactiplantibacillus plantarum SD69.B2 and Saccharomyces cerevisiae SD69.E3 were used as starters, while α-amylase, amyloglucosidase and protease were assessed for their ability to hydrolyze starch and proteins, providing more available carbon and nitrogen sources for the microorganisms. The bread waste-based sourdoughs made by combining protease and L. plantarum SD69.B2 alone or in combination with S. cerevisiae SD69.E3 were selected based on acidification and growth kinetics, and their biochemical, amino acid, and peptide profiles were also characterized demonstrating promising properties. Therefore, they were used, at different percentages, for bread making. Although a slightly acidic pH and a low leavening power, due to the denatured proteins and gelatinized starch in the bread by-products, the texture and sensory analyses of new breads revealed better textural attributes, smell, acidic taste, and overall acceptability compared to the control. The possible reasons behind such features were discussed. The overall results demonstrated that the approach proposed in this study was effective to valorize bread by-products, and it represents a starting point to develop strategies responding to the current perspective of circular economy in food industry., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Declarations of interest none, (Copyright © 2024. Published by Elsevier B.V.)

Oral delivery of peptide therapeutics is limited by degradation by gut proteases like chymotrypsin. Existing databases of peptidases are limited in size and do not enable systematic analyses of protease substrate preferences, especially for non-natural amino acids. Thus, stability optimization of hit compounds is time and resource intensive. To accelerate the stability optimization of peptide ligands, we generated large datasets of chymotrypsin-resistant peptides via mRNA display to create a predictive model for chymotrypsin-resistant sequences. Through analysis of enriched motifs, we recapitulate known chymotrypsin cleavage sites, reveal positionally dependent effects of monomers on peptide cleavage, and report previously unidentified protective and destabilizing residues. We then developed a machine-learning-based model predicting peptide resistance to chymotrypsin cleavage and validated both model performance and the NGS experimental data by measuring chymotrypsin half-lives for a subset of peptides. Finally, we simulated stability predictions on non-natural amino acids through a leucine hold-out model and observed robust performance. Overall, we demonstrate the utility of mRNA display as a tool for big data generation and show that pairing mRNA display with machine learning yields valuable predictions for chymotrypsin cleavage. Expansion of this workflow to additional proteases could provide complementary predictive models that focus future peptide drug discovery efforts., (© 2024 Wiley‐VCH GmbH.)

Microbial inulinase enzymes have a number of applications in biotechnology. In this study, new strains of Aspergillus welwitschiae were investigated as producers of inulinases and their endo- and exo-inulases were characterized in silico and their protein modeling was performed. The inulinase production by A. welwitschiae employing the Dohelert method to assess the interaction between temperature and cultivation humidity, resulted in a fourfold increase in activity. The optimal temperature and humidity were 25-27 °C and 75-82 %, respectively. The enzyme exhibited optimal activity at a pH of 3.5 and at 60 °C. The hydrolysis of the sisal root yielded 272 mg/g of reducing sugars and oligosaccharides. Our in silico experiments predicted 10 well-validated structures of endo- (5) and exo-inulinases (5) from A. welwitschiae. The notable activity of these inulinases on inulin highlights their potential for the treatment of agro-industrial residues with the objective of producing high-value added products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Enzymes encapsulated in extracellular vesicles (EV)s hold promise as biomarkers for early cancer diagnosis. However, precise measurement of their catalytic activities within EVs remains a notable challenge. Here, we report an enzymatically triggered spherical DNA nanomotor (EDM) that enables one-pot, cascaded, and highly sensitive analysis of the activity of EV-associated or free apurinic/apyrimidinic endonuclease 1 (APE1, a key enzyme in base excision repair) across various biological samples. The EDM capitalizes on APE1-triggered activation of DNAzyme (Dz) and its autonomous cleavage of substrates to achieve nonlinear signal amplification. Using EDM, we reveal a strong correlation between APE1 activity in EVs and that of their parental cancer cells. Additionally, EV APE1 mirrors the fluctuation of cellular APE1 activity in response to chemotherapy-induced DNA damage. In a pilot clinical study (n = 63), the EDM-based assay reveals that more than 80% of active APE1 in serum samples is EV-encapsulated. Notably, EV APE1 can differentiate early prostate cancer (PCa) patients from healthy donors (HDs) with an overall accuracy of 92%, outperforming free APE1 in sera. We anticipate that EDM will become a versatile tool for quantifying EV-associated enzymes., (© 2024 Wiley‐VCH GmbH.)

Uncontrolled use or improper disposal of bulk forms and nanoparticles of heavy metals may lead to their release into the environment. Coastal and floodplain ecosystems are particularly vulnerable, and the effects of metal nanoparticles on Fluvisol and Stagnic Fluvisol are poorly studied. This study aims to examine the effect of heavy metals on the enzymatic activity of the soil, the abundance of culturable microorganisms, growth, and antioxidant status of H. vulgare L. A model experiment was carried out with contamination of Stagnic Fluvisol Humic and Fluvisol with 2200 and 1320 mg kg -1 Zn and Cu, to assess the ecotoxicity of bulk forms and nanoparticles of ZnO and CuO in floodplain soils. The abundance of culturable microorganisms, namely copiotrophs, prototrophs, oligotrophs and nitrogen fixers increased. However, a sharp decrease in dehydrogenase activity and denitrification occurred. This effect was more pronounced in Fluvisol (7 times) than in Stagnic Fluvisol Humic (3 times). The accumulation of HMs was also higher in plants grown in Fluvisol (16-32 times) than in Stagnic Fluvisol Humic (13-24 times), which led to a decrease in plant growth and activation of antioxidant defense systems. An increase in the level of malondialdehyde, and the activity of superoxide dismutase and catalase indicates the induction of oxidative stress. Heavy metals have a greater impact on the biological properties of Fluvisol compared to Stagnic Fluvisol Humic. The presence of heavy metals boosts the abundance of culturable microorganisms, while nanoparticles hinder plant growth more than bulk heavy metals., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

From marine to terrestrial environments, Pseudomonas spp. exhibit a remarkable ability not only to adapt but also thrive even amidst adverse conditions. This fact turns Pseudomonas spp. into one of the most prominent candidates for novel biotechnological solutions. Even though terrestrial isolates have been extensively studied, there is still an almost untapped source to be explored in marine Pseudomonas. Harnessing such strains offers an opportunity to discover novel bioactive compounds that could address current global challenges in healthcare and sustainable development. Therefore, this minireview aimed to provide an overview of the main recent discoveries regarding antimicrobials, antifouling, enzymes, pigments, and bioremediation strategies derived from marine isolates of Pseudomonas spp. Future research perspectives will also be discussed to foster forthcoming endeavors to explore the marine counterparts of such a prolific bacterial genus., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

Obesity stands out as one of the most significant health problems in the modern world. The prevalence of high-calorie diets (HCDs) globally exacerbates this condition. Throughout history, plants and plant-derived food products have been utilized for medicinal purposes, demonstrating their efficacy in the treatment and prevention of various diseases. Gundelia tournefortii (GT), a plant of interest, is known to possess beneficial properties. Hence, this study aimed to investigate the immunotoxic and neurotoxic effects of two different doses of GT plant extract on the liver, brain, and heart tissues of obese rats. For this purpose, Wistar male rats were divided into four groups: "CG," "HCDG," "HCDGUN1," and "HCDGUN2" At the conclusion of the study, adenosine deaminase (ADA) and myeloperoxidase (MPO) activities, as well as acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) biomarkers, were evaluated in the liver, heart, and brain tissues. The study results revealed a statistically significant increase in ADA and MPO activities in the HCDG group compared to the CG group, alongside a significant decrease in the HCDGUN groups compared to the HCDG group. Regarding AChE and BChE activities, a statistically significant decrease was observed in the HCDG group compared to the CG group, whereas an increase was noted in the HCDGUN groups relative to the HCDG group, with the latter approaching values similar to those of the control group. In conclusion, the intake of GT plant extract exhibited positive effects on the immunotoxic and neurotoxic effects induced by HCD in rats with an experimental obesity model, as evidenced by tissue biomarker evaluations., (© 2024 The Author(s). Biotechnology and Applied Biochemistry published by Wiley Periodicals LLC on behalf of International Union of Biochemistry and Molecular Biology.)

Stone fruits and their processed products are highly valued in the whole world for their flavor, aroma, rich nutritional contents, and various health benefits. While large quantities of stone fruits are produced globally, significant losses occur due to improper handling and storage, from production to consumption. This review focuses on the application of advanced non-thermal treatment techniques for whole fresh stone fruits and their processed products. It provides a comprehensive assessment of the factors contributing to spoilage, along with the mechanisms, applications, and limitations of non-thermal techniques in reducing spoilage. Compared to traditional preservation methods, such as the use of artificial food additives, chemicals, thermal treatments, and low-temperature storage, these novel techniques demonstrate better results in minimizing spoilage. Moreover, non-thermal techniques are most sustainable and eco-friendly, as they reduce energy consumption, minimize chemical use, and generate less waste than traditional methods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Enzymes play a vital role in baking, providing significant benefits from dough development to extending shelf life, which enhances product quality and consistency. Acting as biological catalysts, enzymes such as proteases and amylases break down proteins and starches, modifying dough rheology and improving fermentation. Lipases and oxidases further refine dough texture through emulsification and oxidation, while lipases also produce fatty acid derivatives during fermentation, contributing to the flavor and aroma of baked goods. Xylanases and cellulases optimize dough handling by altering fiber structure, and amylases help maintain moisture and texture, extending the shelf life of baked products. Ensuring regulatory compliance is essential when incorporating enzymes into baking processes, as bakers must address enzyme stability and determine appropriate dosages for reliable outcomes. Ongoing research is exploring innovative enzyme applications, including customized enzyme blends that target specific product qualities, offering new possibilities for product differentiation and innovation. In summary, enzyme-driven advancements present bakers with opportunities to improve product quality, shelf life, and consistency, while meeting industry regulations. This review emphasizes the critical impact enzymes have on dough properties and finished product characteristics, highlighting their role in driving future innovations within the baking industry., Competing Interests: Declaration of competing interest The authors declared no conflict of interest., (Published by Elsevier B.V.)

Synthetic polymers, such as plastics, have permeated all aspects of modern life, and nowadays plastic pollution is a major environmental problem. Mycodegradation of these polymers could represent part of the solution to this problem since it calls on a broad toolbox of enzymes and applies non-enzymatic mechanisms to degrade and deteriorate recalcitrant materials. However, not enough is known about this ability for most of the representatives of the fungal kingdom. Another bottleneck is the harmonisation of technologies to analyse plastic degradation. This work involved the design of a biodegradation experiment, where the potential of four fungi representative of Dikarya and Penicillia (Funalia floccosa, Trametes versicolor, Pycnoporus cinnabarinus and Penicillium oxalicum) were tested on their ability to deteriorate the six most used plastics based on gravimetry and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). The following correlation between changes in the band signals and the loss of mass after treatment was determined using polyethylene terephthalate, polypropylene, polyethylene, poly vinyl chloride, high density polyethylene, low density polyethylene and nylon. After treatment, the decrease in absorbance of the characteristic bands of the plastics was taken as an indication of the degradation of the corresponding bonds/functionalities. The four fungi used could transform CH, CH 2 , CH 3 , CO, CO, CN, NH and CCl bonds. The best result was obtained using the fungus F. floccosa with 90-day treatments for high density polyethylene (∼ 62.0 %), low density polyethylene (∼ 23.6 %) and nylon (∼ 35.6 %). Therefore, mycodegradation could open up new doors in the fight against plastic pollution., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Edible and medicinal mushrooms possess excellent nutritional properties due to their incredible versatility in growing on different substrates and producing extracellular enzymes with a wide range of specificity. These features make them excellent candidates for various biotechnological applications. In this context, biotechnological applications using edible and medicinal mushrooms can focus on the bioprocessing of agro-industrial wastes, an economical and environmentally friendly strategy. This review, based on recent original research and scientific reviews, highlights the versatility and potential of mushrooms in terms of sustainability and efficiency. We emphasized the biotechnological applications of edible and medicinal mushrooms and their enzymes including food production with high nutraceutical value by enhancing the quality and flavor of food industry products. Other biotechnological applications addressed in this review were cosmeceutical and biomedical development using mushroom extracts with bioactive compounds; wood pulp pretreatment processes in the pulp and paper industry; bioethanol production; and bioremediation for decontaminating soils and polluted effluents. These applications explain how edible and medicinal mushrooms have gained significance in biotechnology over the years, opening new avenues for innovation. The current tendency to study edible and medicinal mushrooms has gained the attention of researchers because these are still less known organisms becoming an attractive and natural source of novel bioactive compounds that could be integrated into a circular model production., (© 2024 Institute of Food Technologists.)

Silver nanoparticles (AgNPs) have drawn a lot of attention from a variety of fields, particularly the biological and biomedical sciences. As a result, it is acknowledged that AgNPs' direct interactions with macromolecules such as DNA, proteins, and enzymes are essential for both therapeutic and nanotoxicological applications. Enzymes as important catalysts may interact with AgNP surfaces in a variety of ways. Therefore, mechanistic investigation into the molecular effects of AgNPs on enzyme conformation and function is necessary for a comprehensive assessment of their interactions. In this overview, we aimed to overview the various strategies for producing AgNPs. We then discussed the enzyme activity inhibition (EAI) mechanism by nanostructured particles, followed by an in-depth survey of the interaction of AgNPs with different enzymes. Furthermore, various parameters influencing the interaction of NPs and enzymes, as well as the antibacterial and anticancer effects of AgNPs in the context of the enzyme inhibitors, were discussed. In summary, useful information regarding the biological safety and possible therapeutic applications of AgNPs-enzyme conjugates may be obtained from this review., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The different parts of Sorbus torminalis (L.) Crantz are used in traditional medicine for various conditions such as cardiac diseases, cough, and diabetes, indicating its significant medicinal potential. Therefore, the current investigation aimed to reveal the phenolic composition of the poorly studied S. torminalis methanol extract of the bark, as well as its capacity to inhibit enzymes relevant to cardiovascular, neurodegenerative, and metabolic diseases. A total of 28 phenolic components, including 20 procyanidins aglycones (A- and B-type), four procyanidin glycosides, catechin and its glycoside, and two (epi)catechin derivatives, were detected using LC-MS. The contents of total polyphenols (6.22 %), total tannins (3.04 %), condensed tannins (0.70 %), and total flavonoids (0.24 %) were determined spectrophotometrically, highlighting the considerable phenolic richness of the examined plant material. The concentration-dependent ability to inhibit α-amylase (IC 50 =130 μg /mL), α-glucosidase (IC 50 =312.13 μg /mL), acetylcholinesterase (IC 50 =156.46 μg /mL), butyrylcholinesterase (IC 50 =217.68 μg /mL), and angiotensin-converting enzyme (IC 50 =36.77 μg /mL) was demonstrated in vitro. The in silico approach showed that catechin, procyanidin B2 and C1, S. torminalis bark constituents, can form stable complexes with the target enzymes but with different binding affinity. The results supported the medicinal potential of S. torminalis bark and significantly expanded our knowledge of its chemistry, justifying further research., (© 2024 Wiley-VHCA AG, Zurich, Switzerland.)

The aim of the presented study was to evaluate the release of the enzymatically initiated production of hexanal from double emulsion electrospun bio-active membranes at a temperature of fruit storage. Among different formulations of water-in-oil (W1/O) primary emulsions, the emulsion composed of 12% w/v Tween20 and 0.1 M NaCl in water (W1) and 6% of poly(glycerol) poly(ricinoleate) dissolved in sunflower oil (O) using W1/O ratio of 80/20 (w/w) (Tween20-NaCl/6% PGPR) was selected, for further incorporation of enzymes, based on the lowest average droplet size (391.0 ± 15.6 nm), low polydispersity index (0.255 ± 0.07), and good gravitational stability also after 14 days. Both enzymes, lipase and lipoxygenase are needed to produce hexanal (up to 58 mg/L). Additionally, double emulsions were prepared with sufficient conductivity and viscosity using different W1/O to W2 ratios for electrospinning. From the selected electrospun membrane, up to 4.5 mg/L of hexanal was released even after 92 days., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Current views of O 2 accumulation in Earth history depict three phases: The onset of O 2 production by ∼2.4 billion years ago; 2 billion years of stasis at ∼1 % of modern atmospheric levels; and a rising phase, starting about 500 million years ago, in which oxygen eventually reached modern values. Purely geochemical mechanisms have been proposed to account for this tripartite time course of Earth oxygenation. In particular the second phase, the long period of stasis between the advent of O 2 and the late rise to modern levels, has posed a puzzle. Proposed solutions involve Earth processes (geochemical, ecosystem, day length). Here we suggest that Earth oxygenation was not determined by geochemical processes. Rather it resulted from emergent biological innovations associated with photosynthesis and the activity of only three enzymes: 1) The oxygen evolving complex of cyanobacteria that makes O 2 ; 2) Nitrogenase, with its inhibition by O 2 causing two billion years of oxygen level stasis; 3) Cellulose synthase of land plants, which caused mass deposition and burial of carbon, thus removing an oxygen sink and therefore increasing atmospheric O 2 . These three enzymes are endogenously produced by, and contained within, cells that have the capacity for exponential growth. The catalytic properties of these three enzymes paved the path of Earth's atmospheric oxygenation, requiring no help from Earth other than the provision of water, CO 2 , salts, colonizable habitats, and sunlight., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Biofuel cells have recently gained popularity as a green and renewable energy source. Biofuel cells are unique devices of energy and are capable of converting the stored chemical energy from waste materials such as pollutants, organics and wastewater into reliable, renewable, pollution-free energy sources through the action of biocatalysts such as various microorganisms and enzymes. It is a promising technological device to treat waste to compensate for global warming and the energy crisis through the green energy production process. Due to their unique properties, various potential biocatalysts are attracting researchers to apply them to various microbial biofuel cells for improving electricity and power. Recent research in biofuel cells is focusing on the exploitation of different biocatalysts and how they are enhancing power generation for various applications in the field of environmental technology, and biomedical fields such as implantable devices, testing kits, and biosensors. This review focusing the importance of microbial fuel cells (MFCs) and enzymatic fuel cells (ECFs) and role of different types of biocatalysts and their mechanisms for improving biofuel cell efficiency gathered from recent reports. Finally, its multifaceted applications with special emphasis on environmental technology and biomedical field will be described, along with future perspectives.

Three glycolytic enzymes phosphofructokinase (PFK), glyceraldehyde-3-phosphate dehydrogenase (GADPH) and pyruvate kinase (PK) that belong to Staphylococcus aureus were used as targets for screening a dataset composed of 7229 compounds of which 1416 were FDA-approved. Instead of catalytic sites, evolutionarily less conserved allosteric sites were targeted to identify compounds that would selectively bind the bacteria's glycolytic enzymes instead of the human host. Seven different allosteric sites provided by three enzymes were used in independent screening experiments via docking. For each of the seven sites, a total of 723 compounds were selected as the top 10 % which displayed the highest binding affinities. All compounds were then united to yield the top 54 drug candidates shared by all seven sites. Next, 17 out of 54 were selected and subjected to in vitro experiments for testing their inhibition capability for antibacterial growth and enzymatic activity. Accordingly, four compounds displaying antibacterial growth inhibition above 40 % were determined as Candesartan cilexetil, Montelukast (sodium), Dronedarone (hydrochloride) and Thonzonium (bromide). In a second round of experiment, Candesartan cilexetil and Thonzonium displayed exceptionally high killing efficiencies on two bacterial strains of S.aureus (methicillin-sensitive and methicillin-resistant) with concentrations as low as 4 μg/mL and 0.5 μg/mL. Yet, their enzymatic assays were not in accordance with their killing effectiveness. Different inhibitory effects was observed for each compound in each enzymatic assay. A more effective target strategy would be to screen for drug compounds that woud inhibit a combination of glycolytic enzymes observed in the glycolytic pathway., (Copyright © 2024. Published by Elsevier Inc.)

Metals are vital in human physiology, which not only act as enzyme catalysts in the processes of superoxide dismutase and glucose phosphorylation, but also affect the redox process, osmotic adjustment, metabolism and neural signals. However, metal imbalances can lead to diseases such as diabetes, which is marked by chronic hyperglycemia and affects wound healing. The hyperglycemic milieu of diabetes impairs wound healing, posing significant challenges to patient quality of life. Wound healing encompasses a complex cascade of hemostasis, inflammation, proliferation, and remodeling phases, which are susceptible to disruption in hyperglycemic conditions. In recent decades, metals have emerged as critical facilitators of wound repair by enhancing antimicrobial properties (e.g., iron and silver), providing angiogenic stimulation (copper), promoting antioxidant activity and growth factor synthesis (zinc), and supporting wound closure (calcium and magnesium). Consequently, research has pivoted towards the development of metal ion-based therapeutics, including innovative formulations such as nano-hydrogels, nano-microneedle dressings, and microneedle patches. Prepared by combining macromolecular materials such as chitosan, hyaluronic acid and sodium alginate with metals, aiming at improving the management of diabetic wounds. This review delineates the roles of key metals in human physiology and evaluates the application of metal ions in diabetic wound management strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A growing number of hydrolase enzymes show promiscuous acyltransferase activity, even under aqueous conditions. Here we report, for the first time, the ability of Pyrobaculum calidifontis VA1 esterase (PestE) to catalyse the formation of a wide range of amides in buffer, where the acyl donor forms a significant structural component in the amide product. The reactions occur under mild conditions and can achieve conversions up to 97% in 6 h for formation of N-benzylfuranamide as the model reaction. We demonstrate PestE's potential in enzyme cascades to make amides from waste PET plastic and the conversion of the terephthalic acid product to tamibarotene, a drug with activity against acute leukemia. Rational mutagenesis led to identification of PestE variants F33L_F289A and F33L. F33L_F289A increased conversion of N-benzylfuranamide by 1.2-fold, and F33L gave a 4-fold increase in conversion to tamibarotene., (© 2024 Wiley‐VCH GmbH.)

Maintaining adequate levels of antiretroviral (ARV) medications is crucial for the efficacy of HIV treatment and prevention regimens. Monitoring ARV levels can predict or prevent adverse health outcomes like treatment failure or drug resistance. However, conventional ARV measurement using liquid chromatography-tandem mass spectrometry (LC-MS/MS) is slow, expensive, and centralized delaying clinical and behavioral interventions. We previously developed a rapid enzymatic assay for measuring nucleotide reverse transcriptase inhibitors (NRTIs) - the backbone of HIV treatment and prevention regimens - based on the drugs' termination of DNA synthesis by HIV reverse transcriptase (RT) enzyme. Here, we expand our work to include non-nucleoside reverse transcriptase inhibitors (NNRTIs) - an ARV class used in established and emerging HIV treatment and prevention regimens. We demonstrate that the REverse Transcriptase ACTivity (REACT) assay can detect NNRTIs including medications used in oral and long-acting/extended-release HIV treatment and prevention. We demonstrate that REACT can measure NNRTIs spiked in either buffer or diluted plasma and that fluorescence can be measured on both a traditional plate reader and an inexpensive portable reader that can be deployed in point-of-care (POC) settings. REACT measured clinically relevant concentrations of five NNRTIs spiked in aqueous buffer. REACT measurements showed excellent agreement between the plate reader and the portable reader, with a high correlation in both aqueous buffer (Pearson's r = 0.9807, P < 0.0001) and diluted plasma (Pearson's r = 0.9681, P < 0.0001). REACT has the potential to provide rapid measurement of NNRTIs in POC settings and may help to improve HIV treatment and prevention outcomes., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

Data on microbial fouling of various synthetic polymer materials, including those used in space technology, are summarized. It has been established that the dominant groups of microbiota of polymer fouling are the genera of mitosporous fungi Aspergillus, Penicillium, Alternaria, Trichoderma. The enzymatic properties of fungal strains from the collection of microbial cultures of the Microbial Depository Center of the National Academy of Sciences of Armenia were studied. It has been shown that Aspergillus fumigatus, Penicillium chrysogenum, P. steckii, Juxtiphoma eupyrena and a number of other fungi have biofouling activity towards polyethylene, polyethylene terephthalate and some other synthetic polymers. New fungal kits have been developed and proposed to evaluate the fungal resistance of polymeric materials. They include fungi isolated from bio-damaged polymers used in space technology and contain 2 to 5 fungal strains instead of 7 to 9 strains in previously used kits. Taking into account the obtained data, a comparative assessment of the fungal resistance of samples of synthetic polymeric materials of various classes that passed accelerated climatic tests has been carried out. It has been established that the kits of biodegradant fungi, composed of cultures of bio-damaged space technology, generally exceeded the activity of the previously used kits, based on which one can judge the obvious advantages of strains isolated from bio-damaged space technology. In the future, these kits could find application not only for biodegradation of polymers, but also for testing the biostability of various polymers, to use for the construction of aviation and space techniques. Moreover, new optimized kits may be developed based on the strains involved in this study., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

The asymmetric synthesis of tetrahydroisoquinolines (THIQs) has gained importance in recent years due to their significant potential in drug development studies. In this study, the conversion of 1-methyl-3,4-dihydroisoquinoline substrate to a chiral amine, 1-methyl-1,2,3,4-tetrahydroisoquinoline, under the catalysis of the stereoselective imine reductase enzyme from Stackebrandtia nassauensis (SnIR) was investigated in detail to elucidate the mechanism and explain the experimental enantioselectivity. The results were found to be in agreement with the experimental data. To elucidate the reaction mechanism, quantum mechanical calculations were performed by considering a large cluster of the active site of the enzyme. In this regard, possible reaction pathways leading to both R- and S-products with the corresponding intermediates and the transition states for the hydride transfer from the cofactor to the substrate were considered by density functional theory (DFT) calculations, and the factors contributing to the observed stereoselectivity were sought. The calculations supported a stepwise mechanism rather than the concerted protonation and the hydride transfer steps. The stereoselectivity in the hydride transfer was found to be due not only to the stability of the enzyme-subtrate complex but also to the corresponding reaction barriers. The calculations were performed at the wB97XD/6-311+G(2df,2p)//B3LYP/6-31G(d,p) level of theory using the PCM approach., (© 2024 The Author(s). ChemPlusChem published by Wiley-VCH GmbH.)

The spatial structure of proteins, largely determined by their amino acid sequences, is also dependent on the environmental conditions under which the folding process takes place. In aqueous environments, exposure of polar amino acids is the driving factor, whereas protein stabilization in amphipathic membranes requires exposure to hydrophobic residues. This observation can be extended to all other environmental conditions under which proteins exhibit biological activity and, most importantly, to the folding process. The fuzzy oil drop (FOD) model assumes a centric location of hydrophobic residues (hydrophobic core) with exposure of polar residues towards the aqueous environment, as the influence of the aqueous environment is extended to include the contribution of other non-aqueous factors, enabling the assessment of their influence on protein structuring. The application of the modified FOD model (FOD-M) we have developed allows the environment to be represented as an external force field in the form of a continuum. The role of environmental conditions allows modification of the funnel model expressing the localization of the energy minimum as dependent on external conditions expressed by the K scale, where K measures the degree of other than polar water factors participating in folding process., Competing Interests: I declare that: This The paper entitled: Protein folding: funnel model revised prepared by Irena Roterman, Mateusz Slupina, and Leszek Konieczny has not been published or presented elsewhere in part or in entirety and is not under consideration by another journal. We have read and understood your journal’s policies, and we believe that neither the manuscript nor the study violates any of these. There are no conflicts of interest to declare., (© 2024 The Authors.)

Background: To remain competitive, brewers must innovate by incorporating novel elements beyond traditional styles. Thus, exploring triticale as a modern substitute for barley malt is promising, especially given its higher amylolytic activity compared to barley. This study aimed to assess the impact of substituting up to 50% of barley malt with unmalted triticale on green beer quality, encompassing multiple stages from wort production to primary fermentation at a laboratory scale., Results: Triticale-based worts (ratios 10-50%) had lower extract content than 100% barley malt. However, incorporating 10% of triticale led to only a 1% decrease in extract content compared to the all-malt wort. Shearzyme® 500L, an endo-1,4-β-xylanase with β-glucanase side activity, effectively addressed wort viscosity by breaking down arabinoxylans and β-glucans in triticale cell walls. All triticale-based beers exhibited lower ethanol content compared to reference beer, as is typical when using adjuncts. In green beer, a 50% triticale ratio lowered ethanol content by 16% (without enzyme) and 19% (with enzyme) compared to 100% malt beer. However, green beer with 10% triticale had satisfactory levels of total polyphenol and vicinal diketone content, among other parameters., Conclusion: Commercial enzyme application significantly enhanced proteolytic activity within the grain. Fermentations of enzyme-treated worts showed higher amino acid levels, further confirming the increased proteolytic activity facilitated by the chosen enzyme. Overall, this study provides a comprehensive analysis of the brewing process using native triticale. Building on this foundation, future studies will focus on optimizing mashing conditions to enhance the fermentation profile of the wort. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Approximately one-third of the entire world's food resources are deemed to be wasted. Palm kernel meal (PKM), a product that is extensively generated by the palm oil industry, exhibits a unique nutrient-rich composition. However, its recycling is seldom prioritized due to numerous factors. To evaluate the impact of enzymatic pretreatment and Lactobacillus plantarum and Lactobacillus reuteri fermentation upon the antioxidant activity of PKM, we implemented integrated metagenomics and metabolomics approaches. The substantially enhanced ( p < 0.05) property of free radicals scavenging, as well as total flavonoids and polyphenols, demonstrated that the biotreated PKM exhibited superior antioxidant capacity. Non-targeted metabolomics disclosed that the Lactobacillus fermentation resulted in substantial ( p < 0.05) biosynthesis of 25 unique antioxidant biopeptides, along with the increased ( p < 0.05) enrichment ratio of the isoflavonoids and secondary metabolites biosynthesis pathways. The 16sRNA sequencing and correlation analysis revealed that Limosilactobacillus reuteri , Pediococcus acidilactici , Lacticaseibacillus paracasei , Pediococcus pentosaceus , Lactiplantibacillus plantarum , Limosilactobacillus fermentum , and polysaccharide lyases had significantly dominated ( p < 0.05) proportions in PMEL, and these bacterial species were strongly ( p < 0.05) positively interrelated with antioxidants peptides. Fermented PKM improves nutritional value by enhancing beneficial probiotics, enzymes, and antioxidants and minimizing anti-nutritional factors, rendering it an invaluable feed ingredient and gut health promoter for animals, multifunctional food elements, or as an ingredient in sustainable plant-based diets for human utilization, and functioning as a culture substrate in the food sector.

Background: Cooling towers (CTs) are crucial to myriad industrial processes, supporting thermal exchange between fluids in heat exchangers using water from lakes and rivers as coolant. However, CT water can sometimes introduce microbial contaminants that adhere to and colonize various surfaces within the CT system. These microorganisms can form biofilms, significantly hindering the system's thermal exchange efficiency. Current treatment strategies employ oxidizing biocides to prevent microbial growth. However, despite their affordability, they do not eliminate biofilms effectively and can lead to corrosive damage within the system. Herein, we aim to devise an anti-biofilm strategy utilizing hydrolytic enzymes (such as α-amylase, glucoamylase, pectin-lyase, cellulase, protease, and DNase) alongside antibiotics (including meropenem, ciprofloxacin, gentamicin, erythromycin, chloramphenicol, and ceftriaxone) to combat microbial growth and biofilm formation in cooling systems., Results: All enzymes reduced the development of the biofilms significantly compared to controls (p < 0.05). The polysaccharidases exhibited biomass reduction of 90%, except for pectin-lyase (80%), followed by DNAse and protease at 43% and 49%, respectively. The antibiotics reduced the biofilms of 70% of isolates in concentration of > 2 mg/mL. The minimal biofilm eradication concentration (MBEC) lower than 1 mg/mL was detected for some 7-day-old sessile isolates. The enzymes and antibiotics were also used in combination against biofilms using the modified Chequerboard method. We found six synergistic combinations, with Fractional inhibitory concentrations (FIC) < 0.5, out of the ten tested. In the presence of the enzymatic mixture, MBECs presented a significant decrease (p < 0.05), at least 4-fold for antibiotics and 32-fold for enzymes. Moreover, we characterized high molecular weight (> 12 kDa) exopolysaccharides (EPS) from biofilms of ten isolates, and glycosyl composition analysis indicated a high frequency of glucose, mannose, erythrose, arabinose, and idose across isolates EPS contrasting with rhamnose, allose, and those carbohydrates, which were detected in only one isolate., Conclusion: The synergistic approach of combining enzymes with antibiotics emerges as a highly effective and innovative strategy for anti-biofilm intervention, highlighting its potential to enhance biofilm management practices., (© 2024. The Author(s).)

Phenazine natural products play various roles such as signal molecules, antibiotics, or electron carriers in their producer strains. Among these products, phenazinomycin and lavanducyanin, which are produced by Streptomyces species, are characterized by an N-alkyl modification. Herein, we established the biosynthetic pathways for these two phenazine natural products. Gene-disruption experiments and in vitro reconstitution of the phenazine-tailoring pathway revealed the late steps of the biosynthetic pathway of the phenazines. The class II terpene cyclase homolog Pzm1 catalyzes the cyclization reaction of farnesyl diphosphate to form monocyclic farnesyl diphosphate. Additionally, the prenyltransferase homolog PzmP functions as the N-prenyltransferase of 5,10-dihydrophenazine-1-carboxylic acid. The flavin monooxygenase homolog PzmS catalyzes the oxidative decarboxylation of prenylated 5,10-dihydrophenazine-1-carboxylic acid to yield phenazinomycin. This study highlights unprecedented modification enzymes for phenazine natural products., (© 2024 The Author(s). ChemBioChem published by Wiley-VCH GmbH.)

Two-component signal transduction systems (TCSs) are regulatory systems widely distributed in eubacteria, archaea, and a few eukaryotic organisms, but not in mammalian cells. A typical TCS consists of a histidine kinase and a response regulator protein. Functional and mechanistic studies on different TCSs have greatly advanced the understanding of cellular phosphotransfer signal transduction mechanisms. In this concept paper, we focus on the His-Asp phosphotransfer mechanism, the ATP synthesis function, antimicrobial drug design, cellular biosensors design, and protein allostery mechanisms based on recent TCS investigations to inspire new applications and future research perspectives., (© 2024 Wiley-VCH GmbH.)

Background and Objectives: Phase I and phase II drug-metabolizing enzymes are crucial for the metabolism and elimination of various endogenous and exogenous compounds, such as small-molecule hormones, drugs, and xenobiotic carcinogens. While in vitro and animal studies have suggested a link between genetic mutations in these enzymes and an increased risk of cancer, human in vivo studies have provided limited supportive evidence., Methods: Genome-wide association studies (GWASs) are a powerful tool for identifying genes associated with specific diseases by comparing two large groups of individuals. In the present study, we analyzed a GWAS database to identify key diseases genetically associated with drug-metabolizing enzymes, focusing on UDP-glucuronosyltransferases (UGTs)., Results: Our analysis confirmed a strong association between the UGT1 gene and hyperbilirubinemia. Additionally, over ten studies reported a link between the UGT1 gene and increased low-density lipoprotein (LDL) cholesterol levels. UGT2B7 was found to be associated with testosterone levels, total cholesterol levels, and vitamin D levels., Conclusions: Despite the in vitro capability of UGT1 and UGT2 family enzymes to metabolize small-molecule carcinogens, the GWAS data did not indicate their genetic association with cancer, except for one study that linked UGT2B4 to ovarian cancer. Further investigations are necessary to fill the gap between in vitro, animal, and human in vivo data.

Microbial attachment and biofilm formation on microplastics (MPs <5 mm in size) in the environment have received growing attention. However, there is limited knowledge of microbial function and their effect on the properties and behavior of MPs in the environment. In this study, microbial communities in the plastisphere were explored to understand microbial ecology as well as their impact on aquatic ecosystems. Using the amplicon sequencing of 16S and internal transcribed spacer (ITS) genes, we uncovered the composition and diversity of bacterial and fungal communities in samples of MPs (fiber, film, foam, and fragment), surface water, bottom sediment, and coastal sand in two contrasting coastal areas of Japan. Differences in microbial diversity and taxonomic composition were detected depending on sample type (MPs, water, sediment, and sand) and the research site. Although relatively higher bacterial and fungal gene counts were determined in MP fragments and foams from the research sites, there were no significant differences in microbial community composition depending on the morphotypes of MPs. Given the colonization by hydrocarbon-degrading communities and the presence of pathogens on MPs, the complex processes of microbial taxa influence the characteristics of MP-associated biofilms, and thus, the properties of MPs. This study highlights the metabolic functions of microbes in MP-associated biofilms, which could be key to uncovering the true impact of plastic debris on the global ecosystem., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Activities of key enzymes of energy and carbohydrate metabolism (cytochrome c oxidase (COX), lactate dehydrogenase (LDH), aldolase, glucose 6-phosphate dehydrogenase (G6PDH), and 1-glycerophosphate dehydrogenase (1-GPDH)) were studied in pink salmon Oncorhynchus gorbuscha (Walb.) smolts from the White Sea in a cage experiment simulating the transition from a freshwater to a marine environment. A decrease in COX, G6PDH, and 1-GPDH activities and an increase in LDH and aldolase activities were observed in juveniles with an increase in water salinity. Based on the findings, a redistribution of energy substrates between the reactions of aerobic and anaerobic metabolism towards higher anaerobic ATP synthesis was assumed for pink salmon. This may indicate that adaptive mechanisms rearrange metabolism to provide energy for osmoregulation in pink salmon juveniles when the salinity changes in their habitat., (© 2024. Pleiades Publishing, Ltd.)

Site-selective chemical modifications of proteins have emerged as a potent technology in chemical biology, materials science, and medicine, facilitating precise manipulation of proteins with tailored functionalities for basic biology research and developing innovative therapeutics. Compared to traditional recombinant expression methods, one of the prominent advantages of chemical protein modification lies in its capacity to decorate proteins with a wide range of functional moieties, including non-genetically encoded ones, enabling the generation of novel protein conjugates with enhanced or previously unexplored properties. Among these, approaches for dual or multiple modifications of proteins are increasingly garnering attention, as it has been found that single modification of proteins is inadequate to meet current demands. Therefore, in light of the rapid developments in this field, this review provides a timely and comprehensive overview of the latest advancements in chemical and biological approaches for dual functionalization of proteins. It further discusses their advantages, limitations, and potential future directions in this relatively nascent area., (© 2024 The Author(s). Chemistry - A European Journal published by Wiley-VCH GmbH.)

Bacillus species have attracted significant attention in recent years due to their potential for producing various bioactive compounds with diverse functional properties. This review highlights bioactive substances from food-grade Bacillus strains and their applications in functional foods and nutraceuticals. The metabolic capacities of Bacillus species have allowed them to generate a wide range of bioactive substances, including vitamins, enzymes, anti-microbial peptides, and other non-ribosomal peptides. These substances have a variety of positive effects, including potential cholesterol-lowering and immune-modulatory qualities in addition to anti-oxidant and anti-bacterial actions. The uses and mechanisms of action of these bioactive chemicals can be used to improve the functional qualities and nutritional profile of food products. Examples include the use of anti-microbial peptides to increase safety and shelf life, as well as the use of Bacillus-derived enzymes in food processing to improve digestibility and sensory qualities. The exploitation of bioactive compounds from food-grade Bacillus strains presents a promising frontier in the development of functional foods and nutraceuticals with enhanced health benefits. Due to their wide range of activity and applications, they are considered as important resources for the development of novel medications, agricultural biocontrol agents, and industrial enzymes. Ongoing research into the biosynthetic pathways, functional properties, and applications of these compounds is essential to fully realize their potential in the food industry. This review underscores the significance of various bioactive compounds generated from Bacillus in tackling global issues like environmental sustainability, sustainable agriculture, and antibiotic resistance. Future developments in microbiology and biotechnology will enable us to fully utilize the potential of these amazing microbes, resulting in novel approaches to industry, agriculture, and health. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

In recent years, there has been a growing interest in understanding the enzymatic machinery responsible for the modifications of prenyl side chains and elucidating their roles in natural product biosynthesis. This interest stems from the pivotal role such modifications play in shaping the structural and functional diversity of natural products, as well as from their potential applications to synthetic biology and drug discovery. In addition to contributing to the diversity and complexity of natural products, unique modifications of prenyl side chains are represented by several novel biosynthetic mechanisms. Representative unique examples of epoxidation, dehydrogenation, oxidation of methyl groups to carboxyl groups, unusual C-C bond cleavage and oxidative cyclization are summarized and discussed. By revealing the intriguing chemistry and enzymology behind these transformations, this comprehensive and comparative review will guide future efforts in the discovery, characterization and application of modifications of prenyl side chains in natural product biosynthesis., (© 2024 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Herbivorous insects depend on the host plant to optimize their overall reproductive success, and balanced fertilization may alter the plant's quality against herbivory. Life history traits of the Liriomyza trifolii (Burgess) were determined under laboratory conditions using either unfertilized and fertilized plants of bean [Phaseolus vulgaris L. (Fabaceae)], chrysanthemum [Chrysanthemum × morifolium (Asteraceae)], potato [Solanum tuberosum (Solanaceae)], bell pepper [Capsicum annuum (Solanaceae)], and tomato [Solanum lycopersicum (Solanaceae)]. Results indicated that L. trifolii completed development on all studied unfertilized and fertilized plants. Nevertheless, a higher performance of the leaf miner was observed on bean and bell pepper plants compared to the other plants. Furthermore, there was an interaction of the host plant and fertilization with Calcium Aria or Sitam negatively affecting the fitness-related traits of the leaf miner. Application of these fertilizers resulted in delayed immature development of L. trifolii, decreased survival rate, and reduced adult longevity and fecundity. The activity of cinnamyl alcohol dehydrogenase (CAD), peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) enzymes, as well as phenolic, flavonoid, and lignin content were higher in Calcium Aria + Sitam fertilized plants, intermediate in Calcium Aria and Sitam treated plants, and the lower in unfertilized plants. The development and survival of L. trifolii on different host plants, considering fertilization options, become important for deploying cultural control practices against this important pest species., (© 2024. Sociedade Entomológica do Brasil.)

Introduction and Aim: Vitamin D supplementation in aging subjects manifests a positive effect on various health-related parameters. We performed a functionally-histological analysis of the adrenal cortex regarding the factors of vitamin D activity and corticosterone output after vitamin D3 application in a rat model of the andropause., Material and Methods: Middle-aged Wistar rats were divided into sham operated (SO; n=8), orchidectomized (Orx; n=8) and vitamin D3-treated orchidectomized (Orx+vit. D; n=8) groups. Vitamin D3 (5 μg/kg b.m.) was administered subcutaneously for three weeks, while the SO and Orx groups received the vehicle alone. Set objectives were achieved using histochemistry/immunohistochemistry, stereology, ultrastructural and biochemical analyses., Results: Orchidectomy (Orx) decreased the adrenal cortex-related volume densities of vascular (p<0,0001), vitamin D receptor (VDR; p<0,0166), cytochrome P450 oxidase 2R1 (CYP 2R1; p<0,0001) and cytochrome P450 oxidase 24 (CYP 24; p<0,0001) depots, but increased the volume density of cytochrome P450 27B1 (CYP 27B1; p<0,0001) depots. In Orx+vit. D rats, increase of the adrenal cortex-related volume densities of collagen (p<0,0001), VDR (p<0,0001) and CYP 2R1 (p<0,0001) depots as well as the lipid-droplet diameter (p<0,0001) in adrenocortical outer zona fasciculata cells was observed, while a decrease of volume densities of the vascular (p<0,0001), CYP 27B1 (p<0,0001) and CYP 24 (p<0,0001) depots was registered, all versus Orx group. Plasma level of ACTH was decreased (p=0,0155) and serum concentrations of 25-hydroxyvitamin D3 and corticosterone were increased (p<0,0001 and p=0,0187, respectively), all after the same treatment., Conclusions: Increased corticosterone output after vitamin D3 application to andropausal rats appears not to be related to increased availability of 25-hydroxyvitamin D3 and decreased degradation of 1,25-dihydroxyvitamin D3 in adrenal tissue, but rather involves the central regulatory mechanisms., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Live food in the initial life stages of neotropical fish is essential for their development and health; however, it can significantly increase production costs. This study uses complete diets with varying moisture contents (47%, 35%, 24%, and 8%) as a cost-effective alternative in the co-feeding phase of surubim larvae, assessing their influence on development, digestive enzymes, and liver metabolism. In a completely randomized design, 3200 three-day-old Pseudoplatystoma sp. larvae (0.001 g) were distributed evenly among 16 aquariums (20 L), with 200 individuals per aquarium. For the first five days, all larvae were fed Artemia exclusively, after which they were fed experimental diets with varying levels of humidity (47%, 35%, 24%, and 8%) six times a day across four treatments and four replicates. The 21-day feeding trial demonstrated that larvae fed with 24% and 8% moisture diets exhibited increased (p < 0.05) weight gain, final length, and protein efficiency rate. The remaining growth parameters (i.e., specific growth rate and condition factor) did not show significant differences (p > 0.05) among the dietary treatments. Enzymatic analysis revealed that the 47% moisture diet enhanced the amylase and alkaline phosphatase activities, whereas the 24% and 35% moisture diets elevated the lipase and protease activities. The 47% moisture diet also resulted in increased alanine aminotransferase, aspartate aminotransferase, and albumin levels, along with visible hepatic histopathologies in samples, such as visible lipid vacuoles, displacement of the nucleus of the hepatocyte, and increased sinusoid spaces. No significant liver changes were observed in fish fed with other diets. Principal component analysis showed that diets with 8-24% moisture content were the most beneficial during the co-feeding phase of surubim larviculture., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Recent work from Nguyen et al. unveils massively parallel measurements of epistatic interactions between two enzymes, dihydrofolate reductase and thymidylate synthase, in their natural cellular context. Almost 3000 mutations of DHFR in three TYMS backgrounds reveal a complex interaction network. The authors capture much of this complexity using a simple model., Competing Interests: Declaration of interests The author declares no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background Gram-negative Pseudomonas aeruginosa is a common bacteria that is well-known for its capacity to build biofilms, which are organized cell communities encased in a self-produced polymeric matrix. Treating infections becomes more challenging due to biofilms' capacity to provide immunity and resistance to antibiotics. The search for novel anti-biofilm agents has gained significant momentum, and the diverse range of bioactive compounds found in natural products offers a promising avenue. Rich in vitamins, antioxidants, and various phytochemicals, Actinidia deliciosa (kiwi fruit) has demonstrated potential as an antibacterial agent. Aim of the study This study aims to assess the efficacy of A. deliciosa extract in inhibiting biofilm formation by P. aeruginosa in vitro, providing valuable insights into its potential as a natural therapeutic agent for preventing recurrent bacterial infections. Materials and methodology The antibacterial and antibiofilm properties of A. deliciosa (kiwi fruit) methanolic extract were assessed in this study against P. aeruginosa (PAO1). The fruit was gathered, examined by a botanist for authenticity, and then cold macerated in methanol to create an extract. A two-fold broth dilution procedure was used to calculate the minimum inhibitory concentration (MIC), and agar well-diffusion was used to evaluate the antimicrobial activity. Pyocyanin pigment quantification was carried out after the extract was applied, and the antibiofilm impact was evaluated using a crystal violet assay. GraphPad Prism (GraphPad Software, San Diego, CA) was used for statistical analysis. Results Based on our findings, A. deliciosa was shown to have significant antibacterial and antibiofilm properties against P. aeruginosa (PAO1). At 5 mg/mL, the methanolic extract of A. deliciosa exhibited antibacterial activity with an 8 mm zone of inhibition and suppressed PAO1 growth. At 2.5 mg/mL and 1.25 mg/mL doses, PAO1 significantly decreased the production of biofilms by 60% and 29%, respectively. In addition, pyocyanin pigment synthesis was decreased by 30% and 9.25%, respectively, at sub-MIC doses of 2.5 mg/mL and 1.25 mg/mL. When evaluated at 2.5 mg/mL, the extract did not, however, appreciably affect bacterial growth. Conclusion This study enhances the understanding of antibiotic resistance, alternative treatments, and pathogenic microbes. The in vitro findings suggest that A. deliciosa fruit extract may inhibit pyocyanin production in PAO1. Further research with different formulations is recommended to explore its anti-biofilm properties and potential pharmacological applications., Competing Interests: Human subjects: All authors have confirmed that this study did not involve human participants or tissue. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Mathew et al.)

Osteoarthritis (OA), a disease with a multifactorial aetiology and an enigmatic root cause, affects the quality of life of many elderly patients. Even though there are certain medications utilised to reduce the symptomatic effects, a reliable treatment method to reverse the disease is yet to be discovered. Zinc is a cofactor of over 3000 proteins and is the only metal found in all six classes of enzymes. We explored zinc’s effect on the immune system and the bones as OA affects both. We also discussed zinc-dependent enzymes, highlighting their significant role in the disease’s pathogenesis. It is important to note that both excessive and deficient zinc levels can negatively affect bone health and immune function, thereby exacerbating OA. The purpose of this review is to offer a better understanding of zinc’s impact on OA pathogenesis and to provide clarity regarding its beneficial and detrimental outcomes. We searched thoroughly systematic reviews, meta-analysis, review articles, research articles and randomised controlled trials to ensure a comprehensive review. In brief, using zinc supplementation in the treatment of OA may act as a doubled-edged sword, offering potential benefits but also posing risks.

Textile waste is mostly incinerated because few recycling processes are available to recover valuable materials. In this work, a feasible chemo-enzymatic recycling process of wool/polyethylene terephthalate (PET)/elastane blends to recover pure PET is for the first time successfully demonstrated. Two novel enzyme formulations were selected for wool hydrolysis, whereas the recovered amino acids were quantified using high-performance liquid chromatography and two assays (Ninhydrin and Folin-Ciocalteu). Kinetic studies on the amino acid formation alongside reaction observations by scanning electron microscopy proved sufficient removal of wool within 8 hours with the new enzyme formulation, marking an acceleration compared to previous studies. Finally, elastane was separated with a non-hazardous solvent to obtain pure PET. Tensile tests on the recovered PET fibres reveal only slight changes through the enzymatic treatment and no changes induced by the applied solvent. The enzyme formulation was successfully tested on five different post-consumer wool/PET textile waste samples. This valorization approach enhances the circular economy concept for textile waste recycling., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

The aim of this study is to determine the effect of the digestibility of cassava starch by the enzymes extracted from corn malt, which will constitute one of the answers to the problem of integrating local products into the process in a modern brewery. Cassava starch solutions of different concentrations (E0: 0 g/L; E1: 1 g/L; E2: 1.1 g/L; E3: 1.2 g/L; E4: 1.3 g/L; E5: 1.4 g/L and E6: 1.5 g/L) were prepared and subjected to two treatments (gelatinized and non-gelatinized) and 5 mL of each were placed in a test tube. Three millilitres (3 mL) of the solution containing amylases extracted from malt corn was then added to each of the test tubes containing the cassava flour solutions. All the treatments were subjected to three temperature stages (50 °C for 15 min, 90 °C for 20 min, and 100 °C for 75 min). Twenty-eight (28) objects (two duplicates) were experimented in a complete factorial design (2 treatments × 2 temperature levels). The results obtained showed that gelatinization had no effect, which could be due to the high optimum temperatures of corn enzyme activity. The concentrations also did not have significant differences which shows that these concentrations can well be used on an industrial scale to digest cassava starch by corn malt enzymes., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In this work, a horseradish peroxidase (HRP)-encapsulated metal organic framework (MOF)@MOF nanocomposite is developed for detecting H 2 O 2 converted by dismutation of superoxide anions released from live HeLa mitochondria. Initially, an HRP-polyacrylic acid cluster is incorporated on a mesoporous, peroxidase-like Cu/Co-1,4-benzenedicarboxylate (BDC) MOF platform to avoid structural change and deactivation of HRP through its interactions with MOF metal ions. Additionally, a Cu/Co-BDC(HRP)@1,3,5-benzenetricarboxyate (BTC) core-shell MOF/MOF structure, also with peroxide-like properties, serves as a protective matrix for HRP. Then, ultrathin porous carbon shells (UPCS) are adopted to improve the electrical conductivity of the MOF@MOF. The Cu/Co-BDC(HRP)@BTC|UPCS sensing platform exhibits two linear ranges of 0.05-1 µM and 1-1000 µM with a sensitivity of 172 mA mM -1 cm -2 and 1.63 mA mM -1 cm -2 , respectively. A limit of detection of 0.057 µM, good selectivity and stability over 35 days for H 2 O 2 detection are also achieved. After treating the mitochondrial complex with specific inhibitors, amperometric results at the sensing platform confirmed complex I and III within mitochondria as the main electron leakage sites in the electron transfer chains. Therefore, this sensing platform provides a tool that may aid in predicting and even developing treatments for some oxidative stress diseases caused by mitochondrial abnormalities., (© 2024 The Author(s). Small Methods published by Wiley‐VCH GmbH.)

The application of biocatalysis has become essential in both academic and industrial domains for the asymmetric synthesis of chiral amines, and it serves as an alternative tool to transition-metal catalysis and complements traditional chemical methods. It relies on the swift expansion of available processes, primarily as a result of advanced tools for enzyme discovery, combined with high-throughput laboratory evolution techniques for optimizing biocatalysts. This concept paper explores the utilization of non-conventional media such as ether-type solvents, deep eutectic solvents, and micellar catalysis to enhance biocatalytic reactions for chiral amine synthesis. Each section focuses on the unique properties of these media, including their ability to stabilize enzymes, alter substrate solubility, and modulate enzyme selectivity. The paper aims to provide insights into how these innovative media can overcome traditional limitations, offering new avenues for sustainable and efficient chiral amine production through biocatalytic processes., (© 2024 The Author(s). Chemistry - A European Journal published by Wiley-VCH GmbH.)

Introduction: Bacterial surface display is a valuable biotechnology technique for presenting proteins and molecules on the outer surface of bacterial cells. However, it has limitations, including potential toxicity to host bacteria and variability in display efficiency. To address these issues, we investigated the removal of abundant non-essential outer membrane proteins (OMPs) in E. coli as a new strategy to improve the surface display of recombinant proteins., Methods: We targeted OmpA, a highly prevalent OMP in E. coli, using the lambda red method. We successfully knocked out ompA in two E. coli strains, K-12 MG1655 and E. coli BL-21, which have broad research and therapeutic applications. We then combined ompA knockout strains and two OMPs with three therapeutic proteins including an anti-toxin enzyme (ClbS), interleukin 18 (IL-18) for activating cytotoxic T cells and an anti- CTLA4 nanobody (αCTLA4) for immune checkpoint blockade., Results: A total of six different display constructs were tested for their display levels by flow cytometry, showing that the ompA knockout strains increased the percentage as well as the levels of display in bacteria compared to those of isogenic wild-type strains., Conclusions: By removing non-essential, highly abundant surface proteins, we develop an efficient platform for displaying enzymes and antibodies, with potential industrial and therapeutic applications. Additionally, the enhanced therapeutic efficacy opens possibilities for live bacteria-based therapeutics, expanding the technology's relevance in the field., Supplementary Information: The online version contains supplementary material available at 10.1007/s12195-024-00819-w., Competing Interests: Competing interestsThe authors declared that they have no conflict of interest., (© The Author(s), under exclusive licence to Biomedical Engineering Society 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Cell membranes are crucial elements in living organisms, serving as protective barriers and providing structural support for cells. They regulate numerous exchange and communication processes between cells and their environment, including interactions with other cells, tissues, ions, xenobiotics, and drugs. However, the complexity and heterogeneity of cell membranes-comprising two asymmetric layers with varying compositions across different cell types and states (e.g., healthy vs. diseased)-along with the challenges of manipulating real cell membranes represent significant obstacles for in vivo studies. To address these challenges, researchers have developed various methodologies to create model cell membranes or membrane fragments, including mono- or bilayers organized in planar systems. These models facilitate fundamental studies on membrane component interactions as well as the interactions of membrane components with external agents, such as drugs, nanoparticles (NPs), or biomarkers. The applications of model cell membranes have extended beyond basic research, encompassing areas such as biosensing and nanoparticle camouflage to evade immune detection. In this review, we highlight advancements in the engineering of planar model cell membranes, focusing on the nanoarchitectonic tools used for their fabrication. We also discuss approaches for incorporating challenging materials, such as proteins and enzymes, into these models. Finally, we present our view on future perspectives in the field of planar model cell membranes.

The xylanase and glucose oxidase (GOD) are easily inactivated, restricting their applicaiton in food and agriculture fields. In this work, xylanase and glucose oxidase (GOD) were encapsulated into cellulose nanocrystals (CNC) stabilized three-layer microcapsules via ionic gelation technique to improve their bioavailability and targeted delivery. Encapsulation efficiency (EE), physicochemical properties, and in vitro releasing of xylanase and GOD encapsulated in microcapsules were investigated. EE of xylanase and GOD reached the highest values (73.34 % and 67.16 %, respectively) at an enzyme concentration of 35 mg/mL. In vitro experiments revealed that cumulative release of both enzymes encapsulated in microcapsules was greater than that of controls in simulated gastric tract (SGT) and simulated intestinal tract (SIT). The release of xylanase increased from 41.62 % (gastric tract) to 77.13 % (intestine tract), and release of GOD increased from 42.63 % to 72.11 %, respectively. Novel hydrogel carriers as enzymes encapsulation system could effectively improve the survival rate of enzymes in harsh environments and could be widely employed in food, feed and other industries., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Enzymes are molecules that play a crucial role in maintaining homeostasis and balance in all living organisms by catalyzing metabolic and cellular processes. If an enzyme's mechanism of action is inhibited, the progression of certain diseases can be slowed or halted, making enzymes a key therapeutic target. Therefore, identifying or developing enzyme inhibitors is essential for treating significant diseases and ensuring plant defense against pathogens. This review aims to compile information on various types of enzyme inhibitors, particularly those that are well studied and beneficial in both human and plant contexts, by analyzing their mechanisms of action and the resulting benefits. Specifically, this review focuses on three different types of enzyme inhibitors that are most studied, recognized, and cited, each with distinct areas of action and potential benefits. For instance, serine enzyme inhibitors in plants help defend against pathogens, while the other two classes-alpha-glucosidase inhibitors and carbonic anhydrase inhibitors-have significant effects on human health. Furthermore, this review is also intended to assist other researchers by providing valuable insights into the biological effects of specific natural or synthetic inhibitors. Based on the current understanding of these enzyme inhibitors, which are among the most extensively studied in the scientific community, future research could explore their use in additional applications or the development of synthetic inhibitors derived from natural ones. Such inhibitors could aid in defending against pathogenic organisms, preventing the onset of diseases in humans, or even slowing the growth of certain pathogenic microorganisms. Notably, carbonic anhydrase inhibitors have shown promising results in potentially replacing antibiotics, thereby addressing the growing issue of antibiotic resistance.

Food quality is adversely affected by physical, chemical, enzymatic, and microbiological reactions, leading to it becoming inedible. Thus, finding alternative methods to preserve foods effectively and extend their shelf life is important. While chemical preservatives have been effective in preventing the growth of harmful pathogens in foods and extending their shelf life, they can also adversely affect consumers' health. For example, nitrites commonly used as preservatives in processed meats have been linked to the development of cancer. This is why researchers, and the food industry are exploring various options to find nontoxic and safe biopreservatives that can be used to preserve food. One such promising option is biopreservatives because they are derived from natural sources, such as plants and insects. This review explores the antimicrobial properties of various biopreservatives, including bacteriocins, polymers, bacteriophages, enzymes, and natural oils, and how they work together to create a synergistic effect in food preservation.