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Search Results (615)

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Keywords = enzymatic kinetics

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14 pages, 1039 KiB  
Article
Improvement of Soluble Expression, Stability, and Activity of Acetaldehyde Lyase by Elastin-like Polypeptides Fusion for Acetoin Production from Acetaldehyde
by Hui Lin, Jiming Zhang, Jie Hu, Lu Ma, Kaili Lai, Chaosong Zheng, Qiuhua Yang and Liaoyuan Zhang
Biomolecules 2025, 15(9), 1216; https://doi.org/10.3390/biom15091216 - 22 Aug 2025
Abstract
To achieve the large-scale, low-cost preparation of acetaldehyde lyase (ALS), elastin-like polypeptides (ELPs) as non-chromatographic purification tags were employed to develop an ELP-ALS fusion protein in Escherichia coli. Induction expression results demonstrated that the ELPs tag efficiently improved the soluble expression of [...] Read more.
To achieve the large-scale, low-cost preparation of acetaldehyde lyase (ALS), elastin-like polypeptides (ELPs) as non-chromatographic purification tags were employed to develop an ELP-ALS fusion protein in Escherichia coli. Induction expression results demonstrated that the ELPs tag efficiently improved the soluble expression of the ALS enzyme. Through two rounds of inverse transition cycling (ITC), highly pure ELP-ALS was obtained with an enzyme recovery rate of 85.77%, outperforming Ni2+-affinity chromatography (66.80%). The comparative analysis of enzymatic properties revealed that ELP fusion markedly improved the stability and substrate tolerance of the ALS enzyme. Kinetic parameter analysis under identical conditions showed that ELP-ALS possessed a Vmax of 15.25 U/mg and a kcat/Km of 73.05 s−1·M−1, representing 1.86-fold and 2.97-fold improvements over His-ALS, respectively. Fed-batch reaction using ELP-ALS and acetaldehyde as biocatalyst and substrate, respectively, yielded 95.92 g/L acetoin with 49.32% increase compared to His-ALS (64.24 g/L). These results demonstrated the application potential of ELP-ALS as a promising biocatalyst for acetoin production from acetaldehyde due to its lower preparation cost, higher biocatalytic efficiency, better stability, and substrate tolerance. Full article
(This article belongs to the Special Issue Industrial Microorganisms and Enzyme Technologies)
11 pages, 852 KiB  
Article
Furthering the Application of a Low-Moisture Anhydrous Ammonia Pretreatment of Corn Stover
by Ming-Hsun Cheng and Kurt A. Rosentrater
Processes 2025, 13(8), 2643; https://doi.org/10.3390/pr13082643 - 20 Aug 2025
Viewed by 133
Abstract
The use of an ammonia fiber expansion pretreatment using low-moisture anhydrous ammonia (LMAA) is a promising strategy for biomass deconstruction, with significant effects on depolymerizing lignin and hemicellulose. An LMAA pretreatment provides several advantages, including compatibility with the high-biomass loading of solids, efficient [...] Read more.
The use of an ammonia fiber expansion pretreatment using low-moisture anhydrous ammonia (LMAA) is a promising strategy for biomass deconstruction, with significant effects on depolymerizing lignin and hemicellulose. An LMAA pretreatment provides several advantages, including compatibility with the high-biomass loading of solids, efficient ammonia recovery, and scalability for industrial operations. In this study, the reactor was revisited and optimized to improve glucan digestibility from corn stover through enzymatic hydrolysis, building on our previous findings that identified limitations in ammonia distribution. The effects of the biomass particle size, the reaction time, and their interaction on glucose yields were investigated to determine their influence on the subsequent enzymatic hydrolysis kinetics. The best glucose yield of 83% was achieved using an LMAA pretreatment of biomass with a 0.5 mm particle size, representing an improvement of approximately 5% compared to biomass with a 1 mm particle size. Additionally, reactor optimization led to a 22% improvement in the glucose yield compared to the previous reactor configuration. According to the results of the reaction kinetics fitting, the enzymatic hydrolysis data indicated that the reaction followed a pseudo-first-order model. Full article
(This article belongs to the Special Issue Advances in Solid Waste Treatment and Design (2nd Edition))
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18 pages, 422 KiB  
Article
Effects of Replacing Soybean Meal with Enzymatically Fermented Citric Waste Pellets on In Vitro Rumen Fermentation, Degradability, and Gas Production Kinetics
by Gamonmas Dagaew, Seangla Cheas, Chanon Suntara, Chanadol Supapong and Anusorn Cherdthong
Animals 2025, 15(16), 2351; https://doi.org/10.3390/ani15162351 - 11 Aug 2025
Viewed by 277
Abstract
This study evaluated the effects of replacing SBM with CWYWEP on in vitro rumen fermentation, nutrient degradability, and gas production kinetics. Citric waste was co-fermented with yeast waste and a multi-enzyme complex for 14 days, then sun-dried and pelleted. The final CWYWEP product [...] Read more.
This study evaluated the effects of replacing SBM with CWYWEP on in vitro rumen fermentation, nutrient degradability, and gas production kinetics. Citric waste was co-fermented with yeast waste and a multi-enzyme complex for 14 days, then sun-dried and pelleted. The final CWYWEP product contained 50.4% crude protein (DM basis). A completely randomized design tested seven diets in which SBM was replaced by CWYWEP or non-enzymatic citric waste–yeast waste pellets (CWYWP) at 0%, 33%, 66%, or 100% inclusion. Replacing SBM with CWYWEP significantly increased cumulative gas production at 96 h, with the 100% CWYWEP group achieving 93.7 mL/0.5 g DM—a 14% increase over the control (p < 0.01). Microbial lag time was reduced to 0.17 h vs. 0.28 h in the control (p < 0.05), suggesting faster microbial colonization. The highest in vitro DM degradability (IVDMD) at 48 h was observed in the 100% CWYWEP group (64.5%), outperforming both the SBM control and all CWYWP treatments (p < 0.01). Notably, CWYWEP increased total volatile fatty acids by 5% at 4 h and propionate by 9% at 2 h, while reducing methane production by 5% (p < 0.05). Other parameters, including pH, ammonia nitrogen, organic matter digestibility, and protozoal counts, were unaffected (p > 0.05). In contrast, CWYWP without enzymes showed minimal improvement. These findings indicate that CWYWEP is a promising high-protein alternative to SBM, enhancing fermentation efficiency and reducing methane under in vitro conditions. Further in vivo studies are warranted to validate these effects. Full article
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20 pages, 2213 KiB  
Article
ANCUT1, a Fungal Cutinase MgCl2-Activated by a Non-Essential Activation Mechanism for Poly(ethylene terephthalate) Hydrolysis
by José Augusto Castro-Rodríguez, Karla Fernanda Ramírez-González, Francisco Franco-Guerrero, Andrea Sabido-Ramos, Ilce Fernanda Abundio-Sánchez, Rogelio Rodríguez-Sotres, Adela Rodríguez-Romero and Amelia Farrés
Catalysts 2025, 15(8), 757; https://doi.org/10.3390/catal15080757 - 7 Aug 2025
Cited by 1 | Viewed by 651
Abstract
Plastic waste, particularly poly(ethylene terephthalate) (PET), negatively impacts the environment and human health. Biotechnology could become an alternative to managing PET waste if enzymes ensure the recovery of terephthalic acid with efficiencies comparable to those of chemical treatments. Recent research has highlighted the [...] Read more.
Plastic waste, particularly poly(ethylene terephthalate) (PET), negatively impacts the environment and human health. Biotechnology could become an alternative to managing PET waste if enzymes ensure the recovery of terephthalic acid with efficiencies comparable to those of chemical treatments. Recent research has highlighted the potential of fungal cutinases, such as wild-type ANCUT1 (ANCUT1wt) from Aspergillus nidulans, in achieving PET depolymerization. Fungal cutinases’ structures differ from those of bacterial cutinases, while their PET depolymerization mechanism has not been well studied. Here, a reliable model of the ANCUT1wt was obtained using AlphaFold 2.0. Computational chemistry revealed potential cation-binding sites, which had not been described regarding enzymatic activation in fungal cutinases. Moreover, it allowed the prediction of residues with the ability to interact with a PET trimer that were mutation candidates to engineer the substrate binding cleft, seeking enhancements of PET hydrolysis. Enzyme kinetics revealed that both ANCUT1wt and ANCUT1N73V/L171Q (DM) were activated by MgCl2, increasing the dissociation constant of the substrate and maximal reaction rate. We found that in the presence of MgCl2, DM hydrolyzed different PET samples and released 9.1-fold more products than ANCUT1wt. Scanning Electron Microscopy revealed a different hydrolysis mode of these enzymes, influenced by the polymer’s crystallinity and structure. Full article
(This article belongs to the Special Issue Catalysis Accelerating Energy and Environmental Sustainability)
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13 pages, 1537 KiB  
Article
Correlation of SERPINA-1 Gene Over-Expression with Inhibition of Cell Proliferation and Modulation of the Expression of IL-6, Furin, and NSD2 Genes
by Nassim Tassou, Hajar Anibat, Ahmed Tissent and Norddine Habti
Biologics 2025, 5(3), 22; https://doi.org/10.3390/biologics5030022 - 6 Aug 2025
Viewed by 332
Abstract
Background and Objectives: The cytokine IL-6, methyltransferase NSD2, pro-protein convertase Furin, and growth factor receptor IGF-1R are essential factors in the proliferation of cancer cells. These proteins are involved in the tumor process by generating several cell-signaling pathways. However, the interactions of these [...] Read more.
Background and Objectives: The cytokine IL-6, methyltransferase NSD2, pro-protein convertase Furin, and growth factor receptor IGF-1R are essential factors in the proliferation of cancer cells. These proteins are involved in the tumor process by generating several cell-signaling pathways. However, the interactions of these oncogenic biomarkers, Furin, IL-6, and NSD2, and their links with the inhibitor SERPINA-1 remain largely unknown. Materials and Methods: Cell proliferation is measured by colorimetric and enzymatic methods. The genetic expressions of SERPINA-1, Furin, IL-6, and NSD2 are measured by qRT-PCR, while the expression of IGF-1R on the cell surface is measured by flow cytometry. Results: The proliferation of cells overexpressing SERPINA-1 (JP7pSer+) is decreased by more than 90% compared to control cells (JP7pSer-). The kinetics of the gene expression ratios of Furin, IL-6, and NSD2 show an increase for 48 h, followed by a decrease after 72 h for the three biomarkers in JP7pSer+ cells compared to JP7pSer- cells. The expression of IGF-1R on the cell surface in both cell lines is low, with JP7pSer- cells expressing 1.33 times more IGF-1R than JP7pSer+ cells. Conclusions: These results suggest gene correlations of SERPINA-1 overexpression with decreased cell proliferation and modulation of gene expression of Furin, IL-6, and NSD2. This study should be complemented by molecular transcriptomic and proteomic experiments to better understand the interaction of SERPINA-1 with IL-6, Furin, and NSD2, and their effect on tumor progression. Full article
(This article belongs to the Topic Advances in Anti-Cancer Drugs: 2nd Edition)
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35 pages, 2193 KiB  
Review
How Mechanistic Enzymology Helps Industrial Biocatalysis: The Case for Kinetic Solvent Viscosity Effects
by Gabriel Atampugre Atampugbire, Joanna Afokai Quaye and Giovanni Gadda
Catalysts 2025, 15(8), 736; https://doi.org/10.3390/catal15080736 - 1 Aug 2025
Viewed by 711
Abstract
Biocatalysis is one of the oldest fields that has been used in industrial applications, with one of the earliest purposeful examples being the mass production of acetic acid from an immobilized Acinetobacter strain in the year 1815. Efficiency, specificity, reduced reaction times, lower [...] Read more.
Biocatalysis is one of the oldest fields that has been used in industrial applications, with one of the earliest purposeful examples being the mass production of acetic acid from an immobilized Acinetobacter strain in the year 1815. Efficiency, specificity, reduced reaction times, lower overall costs, and environmental friendliness are some advantages biocatalysis has over conventional chemical synthesis, which has made biocatalysis increasingly used in industry. We highlight three necessary fields that are fundamental to advancing industrial biocatalysis, including biocatalyst engineering, solvent engineering, and mechanistic engineering. However, the fundamental mechanism of enzyme function is often overlooked or given less attention, which can limit the engineering process. In this review, we describe how mechanistic enzymology benefits industrial biocatalysis by elucidating key fundamental principles, including the kcat and kcat/Km parameters. Mechanistic enzymology presents a unique field that provides in-depth insights into the molecular mechanisms of enzyme activity and includes areas such as reaction kinetics, catalytic mechanisms, structural analysis, substrate specificity, and protein dynamics. In line with the objective of protein engineering to optimize enzyme activity, we summarize a range of strategies reported in the literature aimed at improving the product release rate, the chemical step of catalysis, and the overall catalytic efficiency of enzymes. Further into this review, we delineate kinetic solvent viscosity effects (KSVEs) as a very efficient, cost-effective, and easy-to-perform method to probe different aspects of enzyme reaction mechanisms, including diffusion-dependent kinetic steps and rate-limiting steps. KSVEs are cost-effective because simple kinetic enzyme assays, such as the Michaelis–Menten kinetic approach, can be combined with them without the need for specialized and costly equipment. Other techniques in protein engineering and genetic engineering are also covered in this review. Additionally, we provide information on solvent systems in enzymatic reactions, details on immobilized biocatalysts, and common misconceptions that misguide enzyme design and optimization processes. Full article
(This article belongs to the Special Issue Enzyme Engineering—the Core of Biocatalysis)
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24 pages, 3523 KiB  
Article
Mechanistic Elucidation and Establishment of Drying Kinetic Models of Differential Metabolite Regulation in Rheum palmatum During Natural Sun Drying: An Integrated Physiology, Untargeted Metabolomics, and Enzymology Study
by Wen Luo, Jinrong Guo, Jia Zhou, Mingjun Yang and Yonggang Wang
Biology 2025, 14(8), 963; https://doi.org/10.3390/biology14080963 - 1 Aug 2025
Viewed by 303
Abstract
Rhubarb, a medicinal herb in Gansu Province, China, undergoes significant quality changes during sun-drying. This study investigated color changes, drying kinetics, anthraquinone (AQ) content, metabolic profiles, and enzyme activity during the process. Results showed that drying induced enzymatic browning, with the browning index [...] Read more.
Rhubarb, a medicinal herb in Gansu Province, China, undergoes significant quality changes during sun-drying. This study investigated color changes, drying kinetics, anthraquinone (AQ) content, metabolic profiles, and enzyme activity during the process. Results showed that drying induced enzymatic browning, with the browning index (BI) progressively increasing over extended drying periods (4–16 h) and with greater slice thickness (2–8 mm). Catalase (CAT) activity first decreased and then increased, while polyphenol oxidase (PPO) activity decreased throughout drying. Slice thickness significantly affected AQ content, with the highest in 2 mm slices and the lowest in 4 mm slices. The drying process followed a logarithmic model (R2 = 0.99418, RMSE = 0.02310, and χ2 = 0.0005). Metabolomics analysis identified 631 differential metabolites, with 8 key metabolites linked to flavonoid biosynthesis, phenylalanine biosynthesis, and tyrosine metabolism. Fifteen enzymes were involved in metabolite synthesis and decomposition, though some enzyme activity trends contradicted metabolite changes. This study provides insight into rhubarb drying mechanisms and a basis for optimizing the drying process. Full article
(This article belongs to the Section Plant Science)
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11 pages, 471 KiB  
Article
The Intestinal Mechanisms in the Excretion of Pepsinogen, Amylase and Lipase in Coprofiltrate in Women During Pregnancy and the Postpartum Period
by Elena Kolodkina and Sergey Lytaev
Biomolecules 2025, 15(8), 1099; https://doi.org/10.3390/biom15081099 - 29 Jul 2025
Viewed by 307
Abstract
Background: Enzymes secreted by the digestive glands are excreted from the body with urine, sweat and feces, and they are also removed from the blood due to their participation in the enzymatic provision of the secretion entering the gastrointestinal tract. Objective: [...] Read more.
Background: Enzymes secreted by the digestive glands are excreted from the body with urine, sweat and feces, and they are also removed from the blood due to their participation in the enzymatic provision of the secretion entering the gastrointestinal tract. Objective: The aim of this work was to analyze the activity of pepsinogen, amylase and lipase in the coprofiltrate of pregnant women in each trimester of pregnancy and in the postpartum period, taking into account the timing and type of delivery (term, premature, late delivery or cesarean section). Methods: Data from studies of non-pregnant (n = 45) and pregnant (n = 193) women were analyzed. The materials for preparation coprofiltrate were collected during delivery. Pepsinogen activity was determined by proteolytic activity at pH = 1.5–2.0 using the tyrosine spectrophotometric method, while amylase activity was determined by the amyloclastic method of Karavey, and lipolytic activity was determined by a unified kinetic method using olive oil as a substrate. Outcomes: A small amount of pepsinogen was excreted in the coprofiltrate, and while the level of its excretion increased after childbirth, it remained below the control values. At the same time, an increase in the amylolytic activity of the coprofiltrate was observed in all groups of pregnant women examined from the first to the third trimester of pregnancy. In pregnant women, multidirectional changes in lipase activity were observed depending on the timing and type of delivery. Conclusions: At the end of pregnancy, amylolytic activity increased in all women, and pepsinase activity decreased compared to the indicators of non-pregnant women. No reliable differences were found in the lipolytic activity of the coprofiltrate in pregnant women at the end of pregnancy and the indicators of non-pregnant women. Full article
(This article belongs to the Special Issue Digestive Enzymes in Health and Disease)
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18 pages, 3972 KiB  
Article
Naphthalimide-Based Fluorescent Probe for Portable and Rapid Response to γ-Glutamyl Transpeptidase
by Jinhu Wang, Xianchao Jia, Yihao Zhang, Ye Gao, Lei Zhang, Changgong Meng, Zhaohui Wang and Yang Jiao
Molecules 2025, 30(15), 3174; https://doi.org/10.3390/molecules30153174 - 29 Jul 2025
Viewed by 316
Abstract
γ-Glutamyl transpeptidase (GGT) is overexpressed in a variety of diseases, making it an important diagnostic criterion for diseases. Herein, a new fluorescence probe based on naphthalimide (Glu-MDA) was developed and employed for the rapid detection of GGT in tumor cells or samples. Alkynylated [...] Read more.
γ-Glutamyl transpeptidase (GGT) is overexpressed in a variety of diseases, making it an important diagnostic criterion for diseases. Herein, a new fluorescence probe based on naphthalimide (Glu-MDA) was developed and employed for the rapid detection of GGT in tumor cells or samples. Alkynylated naphthalimide is the fluorescent core for excellent fluorescence response. The covalent bridging of self-immolative short linkers reduces the steric hindrance between probes and enzyme cleavage sites, which leads to improved enzymatic reaction kinetics. Glu-MDA shows a rapid response and excellent selectivity with a detection limit of 0.044 U/L. This allows the efficient detection of GGT levels in solution and cells. Simultaneously, the construction of Glu-MDA pre-stained test strips provided an innovative strategy for the qualitative detection of GGT activity, helping to detect GGT faster, more portably, and cost-effectively in various scenarios. Full article
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16 pages, 3152 KiB  
Article
Enzymatic Modification of Walnut Shell for High-Efficiency Adsorptive Methylene Blue Removal
by Xifeng Lv, Xuejian Zhou, Ruiqi Yang, Di Cai and Wenqiang Ren
Materials 2025, 18(15), 3434; https://doi.org/10.3390/ma18153434 - 22 Jul 2025
Viewed by 281
Abstract
Developing energy-efficient and environmentally benign synthesis protocols is crucial to agricultural waste-based adsorbent preparation. This study prepared novel walnut shell-derived adsorbents by enzymatic modification using a green process, and the as-prepared material was used for methylene blue (MB) removal from wastewater. The results [...] Read more.
Developing energy-efficient and environmentally benign synthesis protocols is crucial to agricultural waste-based adsorbent preparation. This study prepared novel walnut shell-derived adsorbents by enzymatic modification using a green process, and the as-prepared material was used for methylene blue (MB) removal from wastewater. The results showed that under the optimized conditions (100 mg L−1 methylene blue (MB) solution, pH 7, 30 °C, 120 min adsorption time, and 0.14 g adsorbent dosage), WS-1 exhibited an MB removal efficiency of 93.67%, which was only slightly lower than that of WS-2 that was prepared by further carbonization of WS-1 using the low-temperature hydrothermal method (99.01%). Kinetic analysis confirmed WS-1 exhibited pseudo-second-order adsorption kinetics, which were generally similar to those of WS-2. However, the results obtained by the isotherm model followed by the Langmuir model of WS-1 indicated monolayer adsorption involving combined weak chemisorption and physisorption, which was different from the WS-2 (followed the Freundlich model that inferred multilayer chemisorption). In conclusion, this study successfully converted walnut shells, a type of agricultural waste, into functional adsorbents by a novel, simple, and greener enzymatic modification method, thereby achieving dual benefits of waste valorization and wastewater treatment. Full article
(This article belongs to the Section Green Materials)
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34 pages, 1773 KiB  
Review
Participants in Transcription–Replication Conflict and Their Role in Formation and Resolution of R-Loops
by Anastasiia T. Davletgildeeva and Nikita A. Kuznetsov
Int. J. Mol. Sci. 2025, 26(14), 6951; https://doi.org/10.3390/ijms26146951 - 19 Jul 2025
Viewed by 892
Abstract
The DNA of all living organisms is a common matrix for both replication and transcription processes. This sometimes leads to inevitable collisions between DNA replication and transcription machinery. There is plethora of evidence demonstrating that such collisions (or TRCs) are one of the [...] Read more.
The DNA of all living organisms is a common matrix for both replication and transcription processes. This sometimes leads to inevitable collisions between DNA replication and transcription machinery. There is plethora of evidence demonstrating that such collisions (or TRCs) are one of the most common and significant reasons for genomic instability. One of the key outcomes of TRCs is the accumulation of non-canonical DNA secondary structures, including R-loops. R-loops are three-stranded DNA–RNA hybrids with a displaced third single-stranded DNA fragment. Although R-loops are thought to play several functional roles in biological processes, an imbalance in their metabolism has been proven to have severe consequences. In this review, we attempt to summarize the current knowledge of the participants in the process of R-loop regulation in cells, with an emphasis on eukaryotic systems. We also touch upon the conditions favoring TRCs and the possible ways of dealing with these conflicts. Full article
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24 pages, 2613 KiB  
Article
Hierarchical Sensing Framework for Polymer Degradation Monitoring: A Physics-Constrained Reinforcement Learning Framework for Programmable Material Discovery
by Xiaoyu Hu, Xiuyuan Zhao and Wenhe Liu
Sensors 2025, 25(14), 4479; https://doi.org/10.3390/s25144479 - 18 Jul 2025
Viewed by 363
Abstract
The design of materials with programmable degradation profiles presents a fundamental challenge in pattern recognition across molecular space, requiring the identification of complex structure–property relationships within an exponentially large chemical domain. This paper introduces a novel physics-informed deep learning framework that integrates multi-scale [...] Read more.
The design of materials with programmable degradation profiles presents a fundamental challenge in pattern recognition across molecular space, requiring the identification of complex structure–property relationships within an exponentially large chemical domain. This paper introduces a novel physics-informed deep learning framework that integrates multi-scale molecular sensing data with reinforcement learning algorithms to enable intelligent characterization and prediction of polymer degradation dynamics. Our method combines three key innovations: (1) a dual-channel sensing architecture that fuses spectroscopic signatures from Graph Isomorphism Networks with temporal degradation patterns captured by transformer-based models, enabling comprehensive molecular state detection across multiple scales; (2) a physics-constrained policy network that ensures sensor measurements adhere to thermodynamic principles while optimizing the exploration of degradation pathways; and (3) a hierarchical signal processing system that balances multiple sensing modalities through adaptive weighting schemes learned from experimental feedback. The framework employs curriculum-based training that progressively increases molecular complexity, enabling robust detection of degradation markers linking polymer architectures to enzymatic breakdown kinetics. Experimental validation through automated synthesis and in situ characterization of 847 novel polymers demonstrates the framework’s sensing capabilities, achieving a 73.2% synthesis success rate and identifying 42 structures with precisely monitored degradation profiles spanning 6 to 24 months. Learned molecular patterns reveal previously undetected correlations between specific spectroscopic signatures and degradation susceptibility, validated through accelerated aging studies with continuous sensor monitoring. Our results establish that physics-informed constraints significantly improve both the validity (94.7%) and diversity (0.82 Tanimoto distance) of generated molecular structures compared with unconstrained baselines. This work advances the convergence of intelligent sensing technologies and materials science, demonstrating how physics-informed machine learning can enhance real-time monitoring capabilities for next-generation sustainable materials. Full article
(This article belongs to the Special Issue Functional Polymers and Fibers: Sensing Materials and Applications)
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23 pages, 8380 KiB  
Article
Characterizing the Fermentation of Oat Grass (Avena sativa L.) in the Rumen: Integrating Degradation Kinetics, Ultrastructural Examination with Scanning Electron Microscopy, Surface Enzymatic Activity, and Microbial Community Analysis
by Liepeng Zhong, Yujun Qiu, Mingrui Zhang, Shanchuan Wei, Shuiling Qiu, Zhiyi Ma, Mingming Gu, Benzhi Wang, Xinyue Zhang, Mingke Gu, Nanqi Shen and Qianfu Gan
Animals 2025, 15(14), 2049; https://doi.org/10.3390/ani15142049 - 11 Jul 2025
Viewed by 324
Abstract
The objective of this study is to investigate the degradation characteristics of oat grass in the rumen of Mindong goats and changes in microbial community attached to the grass surface. Four healthy male goats, aged 14 months, with permanent rumen fistula, in eastern [...] Read more.
The objective of this study is to investigate the degradation characteristics of oat grass in the rumen of Mindong goats and changes in microbial community attached to the grass surface. Four healthy male goats, aged 14 months, with permanent rumen fistula, in eastern Fujian, were selected as experimental animals. The rumen degradation rate of oat grass was measured at 4, 12, 24, 36, 48, and 72 h using the nylon bag method. Surface physical structure changes in oat grass were observed using scanning electron microscopy (SEM), cellulase activity was measured, and bacterial composition was analyzed using high-throughput 16S rRNA gene sequencing technology. The findings of this study indicate that oat grass had effective degradation rates (ED) of 47.94%, 48.69%, 38.41%, and 30.24% for dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), and acidic detergent fiber (ADF), respectively. The SEM was used to investigate the degradation process of oat grass in the rumen. After 24 h, extensive degradation of non-lignified tissue was observed, resulting in the formation of cavities. At 36 h, significant shedding was observed, and by 72 h, only the epidermis and thick-walled tissue, which exhibited resistance to degradation, remained intact. Surface-attached microorganisms produced β-GC, EG, CBH, and NEX enzymes. The activity of these enzymes exhibited a significant increase between 4 and 12 h and showed a positive correlation with the degradation rate of nutrients. However, the extent of correlation varied. Prevotella and Treponema were identified as key genera involved in the degradation of roughage, with their abundance decreasing over time. Principle Coordinate Analysis (PCOA) revealed no significant differences in the rumen microbial structure across different time points. However, Non-Metric Multidimensional Scaling (NMDS) indicated a discernible diversity order among the samples. According to the Spearman correlation coefficient test, Ruminococcus, Fibrobacter, and Saccharoferments exhibited the closest relationship with nutrient degradation rate and surface enzyme activity, displaying a significant positive correlation. In summary, this study delineates a time-resolved correlative framework linking microbial succession to structural and enzymatic dynamics during oat grass degradation. Full article
(This article belongs to the Section Animal Nutrition)
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24 pages, 5097 KiB  
Article
Non-Monotonic Effect of Substrate Inhibition in Conjunction with Diffusion Limitation on the Response of Amperometric Biosensors
by Romas Baronas
Biosensors 2025, 15(7), 441; https://doi.org/10.3390/bios15070441 - 9 Jul 2025
Viewed by 315
Abstract
The non-monotonic behavior of amperometric enzyme-based biosensors under uncompetitive and noncompetitive (mixed) substrate inhibition is investigated computationally using a two-compartment model consisting of an enzyme layer and an outer diffusion layer. The model is based on a system of reaction–diffusion equations that includes [...] Read more.
The non-monotonic behavior of amperometric enzyme-based biosensors under uncompetitive and noncompetitive (mixed) substrate inhibition is investigated computationally using a two-compartment model consisting of an enzyme layer and an outer diffusion layer. The model is based on a system of reaction–diffusion equations that includes a nonlinear term associated with non-Michaelis–Menten kinetics of the enzymatic reaction and accounts for the partitioning between layers. In addition to the known effect of substrate inhibition, where the maximum biosensor current differs from the steady-state output, it has been determined that external diffusion limitations can also cause the appearance of a local minimum in the current. At substrate concentrations greater than both the Michaelis–Menten constant and the uncompetitive substrate inhibition constant, and in the presence of external diffusion limitation, the transient response of the biosensor, after immersion in the substrate solution, may follow a five-phase pattern depending on the model parameter values: it starts from zero, reaches a global or local maximum, decreases to a local minimum, increases again, and finally decreases to a steady intermediate value. The biosensor performance is analyzed numerically using the finite difference method. Full article
(This article belongs to the Special Issue Novel Designs and Applications for Electrochemical Biosensors)
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21 pages, 3177 KiB  
Article
The Physiological and Biochemical Mechanisms Bioprimed by Spermosphere Microorganisms on Ormosia henryi Seeds
by Meng Ge, Xiaoli Wei, Yongming Fan, Yan Wu, Mei Fan and Xueqing Tian
Microorganisms 2025, 13(7), 1598; https://doi.org/10.3390/microorganisms13071598 - 7 Jul 2025
Viewed by 406
Abstract
The hard-seed coat of Ormosia henryi significantly impedes germination efficiency in massive propagation, while conventional physical dormancy-breaking methods often result in compromised seed vigor, asynchronous seedling emergence, and diminished stress tolerance. Seed biopriming, an innovative technique involving the inoculation of beneficial microorganisms onto [...] Read more.
The hard-seed coat of Ormosia henryi significantly impedes germination efficiency in massive propagation, while conventional physical dormancy-breaking methods often result in compromised seed vigor, asynchronous seedling emergence, and diminished stress tolerance. Seed biopriming, an innovative technique involving the inoculation of beneficial microorganisms onto seed surfaces or into germination substrates, enhances germination kinetics and emergence uniformity through microbial metabolic functions and synergistic interactions with seed exudates. Notably, spermosphere-derived functional bacteria isolated from native spermosphere soil demonstrate superior colonization capacity and sustained bioactivity. This investigation employed selective inoculation of these indigenous functional strains to systematically analyze dynamic changes in endogenous phytohormones, enzymatic activities, and storage substances during critical germination phases, thereby elucidating the physiological mechanisms underlying biopriming-enhanced germination. The experimental results demonstrated significant improvements in germination parameters through biopriming. Inoculation with the Bacillus sp. strain achieved a peak germination rate (76.19%), representing a 16.19% increase over the control (p < 0.05). The biopriming treatment effectively improved the seed vigor, broke the impermeability of the seed coat, accelerated the germination speed, and positively regulated physiological indicators, especially amylase activity and the ratio of gibberellic acid to abscisic acid. This study establishes a theoretical framework for microbial chemotaxis and rhizocompetence in seed priming applications while providing an eco-technological solution for overcoming germination constraints in O. henryi cultivation. The optimized biopriming protocol addresses both low germination rates and post-germination growth limitations, providing technical support for the seedling cultivation of O. henryi. Full article
(This article belongs to the Section Plant Microbe Interactions)
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