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Keywords = enzymatic fingerprinting

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20 pages, 3517 KiB  
Article
Diversity and Functional Potential of Gut Bacteria Associated with the Insect Arsenura armida (Lepidoptera: Saturniidae)
by María Griselda López-Hernández, Reiner Rincón-Rosales, Clara Ivette Rincón-Molina, Luis Alberto Manzano-Gómez, Adriana Gen-Jiménez, Julio Cesar Maldonado-Gómez and Francisco Alexander Rincón-Molina
Insects 2025, 16(7), 711; https://doi.org/10.3390/insects16070711 - 10 Jul 2025
Viewed by 716
Abstract
Insects are often associated with diverse microorganisms that enhance their metabolism and nutrient assimilation. These microorganisms, residing in the insect’s gut, play a crucial role in breaking down complex molecules into simpler compounds essential for the host’s growth. This study investigates the diversity [...] Read more.
Insects are often associated with diverse microorganisms that enhance their metabolism and nutrient assimilation. These microorganisms, residing in the insect’s gut, play a crucial role in breaking down complex molecules into simpler compounds essential for the host’s growth. This study investigates the diversity and functional potential of symbiotic bacteria in the gut of Arsenura armida (Lepidoptera: Saturniidae) larvae, an edible insect from southeastern Mexico, using culture-dependent and metagenomic approaches. Bacterial strains were isolated from different gut sections (foregut, midgut, and hindgut) and cultured on general-purpose media. Isolates were identified through 16S rRNA gene sequencing and genomic fingerprinting. Metagenomics revealed the bacterial community structure and diversity, along with their functional potential. A total of 96 bacterial strains were isolated, predominantly Gram-negative bacilli. Rapidly growing colonies exhibited enzymatic activity, cellulose degradation, and sugar production. Phylogenetic analysis identified eight genera, including Acinetobacter, Bacillus, Enterobacter, Pseudomonas, and others, with significant cellulose-degrading capabilities. Metagenomics confirmed Bacillota as the most abundant phylum. These complementary methods revealed abundant symbiotic bacteria with key metabolic roles in A. armida, offering promising biotechnological applications in enzymatic bioconversion and cellulose degradation. Full article
(This article belongs to the Section Insect Behavior and Pathology)
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21 pages, 1894 KiB  
Article
1H HRMAS NMR Metabolomics for the Characterization and Monitoring of Ripening in Pressed-Curd Ewe’s Milk Cheeses Produced Through Enzymatic Coagulation
by David Castejón, José Segura, Karen P. Cruz-Díaz, María Dolores Romero-de-Ávila, María Encarnación Fernández-Valle, Víctor Remiro, Palmira Villa-Valverde and María Isabel Cambero
Foods 2025, 14(13), 2355; https://doi.org/10.3390/foods14132355 - 2 Jul 2025
Viewed by 376
Abstract
A comprehensive characterization of two pressed-curd cheeses produced from ewe’s milk using enzymatic coagulation—Manchego cheese (with Protected Designation of Origin, PDO) and Castellano cheese (with Protected Geographical Indication, PGI)—was performed throughout the manufacturing process (industrial or traditional) and ripening stages (2, 9, [...] Read more.
A comprehensive characterization of two pressed-curd cheeses produced from ewe’s milk using enzymatic coagulation—Manchego cheese (with Protected Designation of Origin, PDO) and Castellano cheese (with Protected Geographical Indication, PGI)—was performed throughout the manufacturing process (industrial or traditional) and ripening stages (2, 9, 30, 90, and 180 days). Proton high-resolution magic angle spinning nuclear magnetic resonance (1H HRMAS NMR) spectroscopy, combined with Principal Component Analysis (PCA) and cluster analysis, was applied to intact cheese samples. The combination of this spectroscopic technique with chemometric methods allows for the characterization of each type of sheep milk cheese according to its geographical origin and production method (artisanal or industrial), as well as the estimation of ripening time. The results demonstrate that HRMAS NMR spectroscopy enables the rapid and direct analysis of cheese samples, providing a comprehensive profile of their metabolites—a metabolic ‘fingerprint’. Full article
(This article belongs to the Section Dairy)
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20 pages, 3312 KiB  
Article
A Deep Learning-Based Approach for Predicting Michaelis Constants from Enzymatic Reactions
by Yulong Li and Kai Wang
Appl. Sci. 2025, 15(7), 4017; https://doi.org/10.3390/app15074017 - 5 Apr 2025
Viewed by 799
Abstract
The Michaelis constant (Km) is defined as the substrate concentration at which an enzymatic reaction reaches half of its maximum reaction velocity. The determination of Km can be applied to the construction and optimization of metabolic networks. Conventional determinations of Km values based [...] Read more.
The Michaelis constant (Km) is defined as the substrate concentration at which an enzymatic reaction reaches half of its maximum reaction velocity. The determination of Km can be applied to the construction and optimization of metabolic networks. Conventional determinations of Km values based on in vitro experiments are time-consuming and expensive. Although there are a series of computational approaches of determining Km values based on deep learning, the complex biological information in enzymatic reactions still makes it challenging to achieve accurate predictions. In this study, we develop a novel deep learning approach called DLERKm for predicting Km by combining the features of enzymatic reactions including products. We constructed a new enzymatic reaction dataset from the Sabio-RK and UniProt databases for the training and testing of DLERKm, which include the information on substrates, products, enzyme sequences, and Km values. DLERKm utilizes pre-trained language models (ESM-2 and RXNFP), molecular fingerprints, and attention mechanisms to extract enzymatic reaction features for the prediction of Km values. To evaluate the performance of DLERKm, we compared it with a state-of-the-art model (UniKP) on the constructed enzymatic reaction datasets. The model prediction results demonstrate that DLERKm achieved superior prediction performances in terms of the evaluated metrics on the benchmark datasets, where the relative improvements of four metrics (RMSE, MAE, PCC, and R2) were 16.3%, 16.5%, 27.7%, and 14.9%, respectively. Ablation experiments and interpretability analysis demonstrate the importance of considering product information when predicting Km values. In addition, DLERKm exhibits reliable predictive performances for different types of enzymatic reactions. Full article
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32 pages, 8118 KiB  
Article
Impact of Coal Waste Rock on Biological and Physicochemical Properties of Soils with Different Agricultural Uses
by Aleksandra Garbacz, Artur Nowak, Anna Marzec-Grządziel, Marcin Przybyś, Anna Gałązka, Jolanta Jaroszuk-Ściseł and Grzegorz Grzywaczewski
Sustainability 2025, 17(6), 2603; https://doi.org/10.3390/su17062603 - 15 Mar 2025
Viewed by 841
Abstract
During the mining process in mines, a problem arises with the formation of coal post-mining waste, which is waste rock. It is often stored by mines on various types of land to manage the resulting spoil. However, this is not without its impact [...] Read more.
During the mining process in mines, a problem arises with the formation of coal post-mining waste, which is waste rock. It is often stored by mines on various types of land to manage the resulting spoil. However, this is not without its impact on the soil. In this study, we determined the biological and physicochemical properties of rhizosphere soils of the podzolic type, subjected to waste rock reclamation and without the influence of waste rock (control), differing in the type of agricultural use and type of plant cover: field-monocotyledonous (oat cultivation), field-dicotyledonous (buckwheat cultivation), and wasteland covered with very species-poor vegetation. Research has shown that long-term cultivation (buckwheat) contributed to the elimination (leveling out) of the microbial and biochemical differences. The addition of waste rock significantly reduced the number of microorganisms synthesizing siderophore, especially on wasteland (decreased by 1.5 log10/gDW). The abundant presence of the genera Acidocella and Acidphilum, absent in wasteland without waste rock, in the unused soil under the influence of waste rock was strongly associated with the effect of lowering the pH by waste rock in soil not used for agriculture. Increased levels of 77 types of bacteria were observed in samples from buckwheat cultivation compared to wasteland. The number of microorganisms resistant to heavy metals as well as microorganisms capable of producing specific Fe-binding ligands—siderophores—decreased under the influence of waste rock. Moreover, the dehydrogenase activity in long-term cultivation both under the influence of waste rock and without its influence was at a similar level. In contrast, an almost 100-fold decrease in dehydrogenase activity was observed in soils with oat cultivation and a more than 4-fold decrease in acid phosphatase (ACP) and alkaline phosphatase (ALP) activity. These parameters provide an effective system for monitoring soil health, from inexpensive and fast methods to advanced and precise techniques. The results can be applied to solve the problems associated with coal mining wastes by developing methods for their use in soils with long-term agricultural use. Full article
(This article belongs to the Section Hazards and Sustainability)
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10 pages, 590 KiB  
Article
Fingerprint Profile Analysis of Eupolyphaga steleophaga Polypeptide Based on UHPLC-MS and Its Application
by Xin Lai, Hongwei Song, Guangli Yan, Junling Ren and Xijun Wang
Pharmaceuticals 2025, 18(2), 166; https://doi.org/10.3390/ph18020166 - 26 Jan 2025
Viewed by 1259
Abstract
Background and Objectives: As a medicinal and food homologous substance, Eupolyphaga steleophaga is renowned for its potential health benefits, including anti-tumor effects, immune system support, and anti-inflammatory properties. Eupolyphaga steleophaga polypeptides have demonstrated significant biological activity, including the regulation of coagulation and lipid [...] Read more.
Background and Objectives: As a medicinal and food homologous substance, Eupolyphaga steleophaga is renowned for its potential health benefits, including anti-tumor effects, immune system support, and anti-inflammatory properties. Eupolyphaga steleophaga polypeptides have demonstrated significant biological activity, including the regulation of coagulation and lipid metabolism. However, the peptide composition of Eupolyphaga steleophaga requires further clarification to facilitate quality control improvements and a deeper investigation into its pharmacological effects. Therefore, this study aimed to simulate the digestive absorption process of Eupolyphaga steleophaga following oral administration and identify its enzymatic components to enhance quality control. Methods: The digestive absorption process was simulated using artificial gastric fluid and pepsin. A fingerprinting method based on ultrahigh-performance liquid chromatography-mass spectrometry (UHPLC-MS)(Acquire UPLC-Synapt G2-Si HDMS, Waters Corporation, Milford, MA, USA) was developed to identify 63 enzymatic components. The enzymolysis polypeptide fingerprint detection method was used to analyze 10 batches of Eupolyphaga steleophaga sourced from Harbin No. 4 Traditional Chinese Medicine Factory. Chromatographic collection was performed using an ACQUITY UPLC BHE C18 column. Gradient elution was carried out using a mixture of 0.1% formic acid with acetonitrile and 0.1% formic acid with water, with an average flow rate of 0.3 mL/min, a column temperature of 40 °C, and an injection volume of 2 μL. The mass spectrometry (MS) conditions were set as follows: the ion source was operated in positive electrospray ionization (ESI+) mode, with a capillary voltage of 2.8 kV and a sampling cone voltage of 40 V. The ion-source temperature was maintained at 110 °C, while the desolvation temperature was set to 400 °C. The cone gas flow rate was 50 L/h, and the desolvation gas flow rate was 800 L/h. The range for the collection of mass-to-charge ratios (m/z) was between 50 and 1200. Results: The UHPLC-MS method demonstrated high accuracy, repeatability, and stability, successfully identifying 63 enzymatic components of Eupolyphaga steleophaga. Furthermore, polypeptide markers for 63 selected components were identified in all 10 batches of Eupolyphaga steleophaga medicinal materials. This approach was validated by including numerical values such as retention times and peak areas, confirming its reliability for quality control enhancement. Conclusions: This novel UHPLC-MS approach serves as a powerful tool for advancing quality control strategies in veterinary medicine, particularly for animal-derived medicines. It lays a solid foundation for subsequent pharmacological studies of Eupolyphaga steleophaga polypeptides, offering a more reliable means to explore their biological activities and therapeutic potential. Full article
(This article belongs to the Section Natural Products)
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20 pages, 15648 KiB  
Article
Unveiling the Anti-Obesity Potential of Thunder God Vine: Network Pharmacology and Computational Insights into Celastrol-like Molecules
by Siyun Zheng, Hengzheng Yang, Jingxian Zheng, Yidan Wang, Bo Jia and Wannan Li
Int. J. Mol. Sci. 2024, 25(23), 12501; https://doi.org/10.3390/ijms252312501 - 21 Nov 2024
Cited by 1 | Viewed by 1476
Abstract
Obesity, characterized by abnormal or excessive fat accumulation, has become a chronic degenerative health condition that poses significant threats to overall well-being. Pharmacological intervention stands at the forefront of strategies to combat this issue. Recent studies, notably by Umut Ozcan’s team, have uncovered [...] Read more.
Obesity, characterized by abnormal or excessive fat accumulation, has become a chronic degenerative health condition that poses significant threats to overall well-being. Pharmacological intervention stands at the forefront of strategies to combat this issue. Recent studies, notably by Umut Ozcan’s team, have uncovered the remarkable potential of Celastrol, a small-molecule compound derived from the traditional Chinese herb thunder god vine (Tripterygium wilfordii) as an anti-obesity agent. In this research, computational chemical analysis was employed, incorporating the “TriDimensional Hierarchical Fingerprint Clustering with Tanimoto Representative Selection (3DHFC-TRS)” algorithm to systematically explore 139 active small molecules from thunder god vine. These compounds were classified into six categories, with a particular focus on Category 1 molecules for their exceptional binding affinity to obesity-related targets, offering new avenues for therapeutic development. Using advanced molecular docking techniques and Cytoscape prediction models, six representative Celastrol-like molecules were identified, namely 3-Epikatonic Acid, Hederagenin, Triptonide, Triptotriterpenic Acid B, Triptotriterpenic Acid C, and Ursolic Acid. These compounds demonstrated superior binding affinity and specificity toward two key obesity targets, PPARG and PTGS2, suggesting their potential to regulate fat metabolism and mitigate inflammatory responses. To further substantiate these findings, molecular dynamics simulations and MM-PBSA free-energy calculations were applied to analyze the dynamic interactions between these small molecules and the enzymatic active sites of their targets. The results provide robust theoretical evidence that support the feasibility of these molecules as promising candidates for anti-obesity therapies. This study underscores the power of the 3DHFC-TRS algorithm in uncovering bioactive compounds from natural sources, such as thunder god vine, and highlights the therapeutic promise of PPARG and PTGS2 as novel obesity-related targets. Furthermore, it emphasizes the essential role of computational science in expediting drug discovery, paving the way for personalized and precision-based treatments for obesity and heralding a future of more effective healthcare solutions. Full article
(This article belongs to the Special Issue Network Pharmacology: An Emerging Field in Drug Discovery)
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11 pages, 1352 KiB  
Article
Changes in the Main Physicochemical Properties and Electrochemical Fingerprints in the Production of Sea Buckthorn Juice by Pectinase Treatment
by Kaihua Guo
Molecules 2024, 29(5), 1035; https://doi.org/10.3390/molecules29051035 - 28 Feb 2024
Cited by 4 | Viewed by 1680
Abstract
Enzymatic hydrolysis using pectinase is critical for producing high-yield and quality sea buckthorn juice. This study determined the optimal temperature, time, and enzyme dosage combinations to guide manufacturers. A temperature of 60 °C, hydrolysis time of 3 h, and 0.3% enzyme dosage gave [...] Read more.
Enzymatic hydrolysis using pectinase is critical for producing high-yield and quality sea buckthorn juice. This study determined the optimal temperature, time, and enzyme dosage combinations to guide manufacturers. A temperature of 60 °C, hydrolysis time of 3 h, and 0.3% enzyme dosage gave 64.1% juice yield—25% higher than without enzymes. Furthermore, monitoring physicochemical properties reveals enzyme impacts on composition. Higher dosages increase soluble solids up to 15% and soluble fiber content by 35% through cell wall breakdown. However, excessive amounts over 0.3% decrease yields. Pectin concentration also declines dose-dependently, falling by 91% at 0.4%, improving juice stability but needing modulation to retain viscosity. Electrochemical fingerprinting successfully differentiates process conditions, offering a rapid quality control tool. Its potential for commercial inline use during enzymatic treatment requires exploration. Overall, connecting optimized parameters to measured effects provides actionable insights for manufacturers to boost yields, determine enzyme impacts on nutrition/functionality, and introduce novel process analytical technology. Further investigations of health properties using these conditions could expand sea buckthorn juice functionality. Full article
(This article belongs to the Special Issue Nano-Functional Materials for Sensor Applications)
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16 pages, 966 KiB  
Article
Simultaneous Analysis of Organic Acids, Glycerol and Phenolic Acids in Wines Using Gas Chromatography-Mass Spectrometry
by Violeta Garcia-Viñola, Candela Ruiz-de-Villa, Jordi Gombau, Montse Poblet, Albert Bordons, Cristina Reguant and Nicolas Rozès
Foods 2024, 13(2), 186; https://doi.org/10.3390/foods13020186 - 5 Jan 2024
Cited by 13 | Viewed by 3982
Abstract
Fermented beverages, particularly wines, exhibit variable concentrations of organic and phenolic acids, posing challenges in their accurate determination. Traditionally, enzymatic methods or chromatographic analyses, mainly high-performance liquid chromatography (HPLC), have been employed to quantify these compounds individually in the grape must or wine. [...] Read more.
Fermented beverages, particularly wines, exhibit variable concentrations of organic and phenolic acids, posing challenges in their accurate determination. Traditionally, enzymatic methods or chromatographic analyses, mainly high-performance liquid chromatography (HPLC), have been employed to quantify these compounds individually in the grape must or wine. However, chromatographic analyses face limitations due to the high sugar content in the grape must. Meanwhile, phenolic acids, found in higher quantities in red wines than in white wines, are typically analyzed using HPLC. This study presents a novel method for the quantification of organic acids (OAs), glycerol, and phenolic acids in grape musts and wines. The approach involves liquid-liquid extraction with ethyl acetate, followed by sample derivatization and analysis using gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) detection mode. The results indicated successful detection and quantification of all analyzed compounds without the need for sample dilution. However, our results showed that the method of adding external standards was more suitable for quantifying wine compounds, owing to the matrix effect. Furthermore, this method is promising for quantifying other metabolites present in wines, depending on their extractability with ethyl acetate. Fermented beverages, particularly wines, exhibit variable concentrations of organic and phenolic acids, posing challenges in their accurate determination. Traditionally, enzymatic methods or chromatographic analyses, mainly high-performance liquid chromatography (HPLC), have been employed to quantify these compounds individually in the grape must or wine. The approach of this proposed method involves (i) methoximation of wine compounds in a basic medium, (ii) acidification with HCl, (iii) liquid-liquid extraction with ethyl acetate, and (iv) silyl derivatization to analyze samples with gas chromatography-mass spectrometry (GC-MS) in ion monitoring detection mode (SIM). The results indicated successful detection and quantification of all analyzed compounds without the need for sample dilution. However, our results showed that the method of adding external standards was more suitable for quantifying wine compounds, owing to the matrix effect. Furthermore, this method is promising for quantifying other metabolites present in wines, depending on their extractability with ethyl acetate. In other words, the proposed method may be suitable for profiling (targeted) or fingerprinting (untargeted) strategies to quantify wine metabolites or to classify wines according to the type of winemaking process, grape, or fermentation. Full article
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16 pages, 3048 KiB  
Article
Exploring the Release of Elastin Peptides Generated from Enzymatic Hydrolysis of Bovine Elastin via Peptide Mapping
by Jianan Zhang, Yang Liu, Liwen Jiang, Tiantian Zhao, Guowan Su and Mouming Zhao
Molecules 2023, 28(22), 7534; https://doi.org/10.3390/molecules28227534 - 10 Nov 2023
Cited by 2 | Viewed by 2268
Abstract
To enhance the understanding of enzymatic hydrolysis and to accelerate the discovery of key bioactive peptides within enzymatic products, this research focused on elastin as the substrate and investigated the variations in peptide profiles and the production of key bioactive peptides (those exceeding [...] Read more.
To enhance the understanding of enzymatic hydrolysis and to accelerate the discovery of key bioactive peptides within enzymatic products, this research focused on elastin as the substrate and investigated the variations in peptide profiles and the production of key bioactive peptides (those exceeding 5% of the total) and their impacts on the biological activity of the hydrolysates. Through the application of advanced analytical techniques, such as stop-flow two-dimensional liquid chromatography and ultra-high-performance liquid chromatography-tandem mass spectrometry, the research tracks the release and profiles of peptides within elastin hydrolysates (EHs). Despite uniform peptide compositions, significant disparities in peptide concentrations were detected across the hydrolysates, hinting at varying levels of bioactive efficacy. A comprehensive identification process pinpointed 403 peptides within the EHs, with 18 peptides surpassing 5% in theoretical maximum content, signaling their crucial role in the hydrolysate’s bioactivity. Of particular interest, certain peptides containing sequences of alanine, valine, and glycine were released in higher quantities, suggesting Alcalase® 2.4L’s preference for these residues. The analysis not only confirms the peptides’ dose-responsive elastase inhibitory potential but also underscores the nuanced interplay between peptide content, biological function, and their collective synergy. The study sets the stage for future research aimed at refining enzymatic treatments to fully exploit the bioactive properties of elastin. Full article
(This article belongs to the Special Issue Study on Physicochemical Properties of Food Protein, 2nd Edition)
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17 pages, 3452 KiB  
Article
In Silico RNAseq and Biochemical Analyses of Glucose-6-Phosphate Dehydrogenase (G6PDH) from Sweet Pepper Fruits: Involvement of Nitric Oxide (NO) in Ripening and Modulation
by María A. Muñoz-Vargas, Salvador González-Gordo, Jorge Taboada, José M. Palma and Francisco J. Corpas
Plants 2023, 12(19), 3408; https://doi.org/10.3390/plants12193408 - 27 Sep 2023
Cited by 8 | Viewed by 1804
Abstract
Pepper (Capsicum annuum L.) fruit is a horticultural product consumed worldwide which has great nutritional and economic relevance. Besides the phenotypical changes that pepper fruit undergo during ripening, there are many associated modifications at transcriptomic, proteomic, biochemical, and metabolic levels. Nitric oxide [...] Read more.
Pepper (Capsicum annuum L.) fruit is a horticultural product consumed worldwide which has great nutritional and economic relevance. Besides the phenotypical changes that pepper fruit undergo during ripening, there are many associated modifications at transcriptomic, proteomic, biochemical, and metabolic levels. Nitric oxide (NO) is a recognized signal molecule that can exert regulatory functions in diverse plant processes including fruit ripening, but the relevance of NADPH as a fingerprinting of the crop physiology including ripening has also been proposed. Glucose-6-phosphate dehydrogenase (G6PDH) is the first and rate-limiting enzyme of the oxidative phase of the pentose phosphate pathway (oxiPPP) with the capacity to generate NADPH. Thus far, the available information on G6PDH and other NADPH-generating enzymatic systems in pepper plants, and their expression during the ripening of sweet pepper fruit, is very scarce. Therefore, an analysis at the transcriptomic, molecular and functional levels of the G6PDH system has been accomplished in this work for the first time. Based on a data-mining approach to the pepper genome and fruit transcriptome (RNA-seq), four G6PDH genes were identified in pepper plants and designated CaG6PDH1 to CaG6PDH4, with all of them also being expressed in fruits. While CaG6PDH1 encodes a cytosolic isozyme, the other genes code for plastid isozymes. The time-course expression analysis of these CaG6PDH genes during different fruit ripening stages, including green immature (G), breaking point (BP), and red ripe (R), showed that they were differentially modulated. Thus, while CaG6PDH2 and CaG6PDH4 were upregulated at ripening, CaG6PDH1 was downregulated, and CaG6PDH3 was slightly affected. Exogenous treatment of fruits with NO gas triggered the downregulation of CaG6PDH2, whereas the other genes were positively regulated. In-gel analysis using non-denaturing PAGE of a 50–75% ammonium-sulfate-enriched protein fraction from pepper fruits allowed for identifying two isozymes designated CaG6PDH I and CaG6PDH II, according to their electrophoretic mobility. In order to test the potential modulation of such pepper G6PDH isozymes, in vitro analyses of green pepper fruit samples in the presence of different compounds including NO donors (S-nitrosoglutathione and nitrosocysteine), peroxynitrite (ONOO), a hydrogen sulfide (H2S) donor (NaHS, sodium hydrosulfide), and reducing agents such as reduced glutathione (GSH) and L-cysteine (L-Cys) were assayed. While peroxynitrite and the reducing compounds provoked a partial inhibition of one or both isoenzymes, NaHS exerted 100% inhibition of the two CaG6PDHs. Taken together these data provide the first data on the modulation of CaG6PDHs at gene and activity levels which occur in pepper fruit during ripening and after NO post-harvest treatment. As a consequence, this phenomenon may influence the NADPH availability for the redox homeostasis of the fruit and balance its active nitro-oxidative metabolism throughout the ripening process. Full article
(This article belongs to the Special Issue Nitric Oxide in Plant Stress and Physiology)
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19 pages, 6107 KiB  
Article
Chemical Fingerprinting, Aorta Endothelium Relaxation Effect, and Enzymatic Inhibition of Canelo (Drimys winteri J. R. Forst. & G. Forst, (D.C) A. Gray, Family Winteraceae) Fruits
by Ruth E. Barrientos, Javier Romero-Parra, Fredi Cifuentes, Javier Palacios, Néstor Jaime Romero-Jola, Adrián Paredes, Gabriel Vargas-Arana and Mario J. Simirgiotis
Foods 2023, 12(13), 2580; https://doi.org/10.3390/foods12132580 - 1 Jul 2023
Cited by 4 | Viewed by 2569
Abstract
Drimys winteri J.R. Forst. & G. Forst (D.C) G. Gray, var. chilensis (canelo) is an endemic tree from Chile. Since pre-Columbian times, it has produced a fruit known as the canelo pepper, (pimienta de canelo) or Foye pepper, which can be used as [...] Read more.
Drimys winteri J.R. Forst. & G. Forst (D.C) G. Gray, var. chilensis (canelo) is an endemic tree from Chile. Since pre-Columbian times, it has produced a fruit known as the canelo pepper, (pimienta de canelo) or Foye pepper, which can be used as a spice. The chemical and biological analysis of canelo fruits is reported for the first time in this study, that is, its phenolic fingerprinting by UHPLC-PDA- Q-orbitrap MS, the antioxidant activity, the enzymatic inhibitory activity, and its relaxation effects on rat aorta. The proximal composition and the mineral content (Ca: 1.45 ± 0.03 mg/100 g; Mg: 7.72 ± 0.03 mg/100 g; Fe: 4.54 ± 0.21 mg/100 g; Zn: 2.99 ± 0.02 mg/100 g; Mn: 1.08 ± 0.03 mg/100 g; Cu: 0.82 ± 0.02 mg/100 g; K: 53.03 ± 0.20 mg/100 g; Na: 0.087 ± 0.00 mg/100 g) are also reported. The canelo fruits showed a total phenolic content of 57.33 ± 0.82 mg GAE/g dry weight. In addition, the total flavonoid content was 38.42 ± 1.32 mg equivalent of QE/g dry weight. The antioxidant activity was evaluated by employing DPPH and ABTS methods (IC50 of 6.65 ± 0.5 and 9.5 ± 0.05 μg/mL, respectively), ORAC (25.33 ± 1.2 μmol Trolox/g dry plant) and FRAP (45.56 ± 1.32 μmol Trolox/g dry plant). The enzymatic inhibition of acetylcholinesterase, butyrylcholinesterase, and tyrosinase (IC50: 1.94 ± 0.07, 2.73 ± 0.05, and 9.92 ± 0.05 µg extract/mL, respectively) is also reported. Canelo extract led to an 89% relaxation of rat aorta. Our results confirm that D. winteri fruits are a rich source of secondary metabolites and can inhibit enzymes associated with neurodegenerative diseases; the results also suggest that canelo may induce a potentially hypotensive effect in rat aorta. The study demonstrates the medicinal properties of canelo fruit and spice. Full article
(This article belongs to the Section Plant Foods)
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27 pages, 9934 KiB  
Article
Physico-Chemical Characterization, DFT Modeling and Biological Activities of a New Zn (II) Complex Containing Melamine as a Template
by Thouraya Salah, Noureddine Mhadhbi, Ali Ben Ahmed, Besma Hamdi, Najeh Krayem, Mohamed Loukil, Ahlem Guesmi, Lotfi Khezami, Ammar Houas, Naoufel Ben Hamadi, Houcine Naïli and Ferdinando Costantino
Crystals 2023, 13(5), 746; https://doi.org/10.3390/cryst13050746 - 29 Apr 2023
Cited by 15 | Viewed by 2612
Abstract
Single crystals of a new organic–inorganic hybrid compound (C3H7N6)2[ZnCl4]·H2O was synthesized and characterized by X-ray diffraction at room temperature, FT-IR and FT-Raman spectroscopies, optical absorption and photoluminescence behavior. The title compound [...] Read more.
Single crystals of a new organic–inorganic hybrid compound (C3H7N6)2[ZnCl4]·H2O was synthesized and characterized by X-ray diffraction at room temperature, FT-IR and FT-Raman spectroscopies, optical absorption and photoluminescence behavior. The title compound belongs to the triclinic space group P1¯, and in the crystal structure, the inorganic layers are built from tetrachloridozincate anions [ZnCl4]2− and free water molecules, linked together by O–H···Cl hydrogen bonds and halogen···halogen interactions. In addition, Hirshfeld surfaces and 2D fingerprint plots estimate the weak intermolecular interactions accountable for the generation of crystal packing. The optimized geometry, vibrational frequencies and various thermodynamic parameters of the title compound calculated using density functional theory (DFT) methods are in agreement with the experimental values. The theoretical calculations were performed using the DFT method at WB97XD/Lanl2dz basis set levels and we discussed topological analysis of atoms in molecules (AIM) at the BCP point. A detailed interpretation of the IR and Raman spectra were reported. Additionally, the simulated spectrum satisfactorily coincided with the experimental UV-Visible spectrum. A wide band gap exceeding 4 eV of the synthesized compound was recorded. The photoluminescence (PL) was characterized through two bands successively at 453 and 477 nm. Ultimately, antimicrobial activity and enzymatic inhibition assays of the complex were also investigated through microbial strains, agar diffusion method, minimum inhibitory concentration (MIC) determination, lipase and phospholipase A2 inhibition. Full article
(This article belongs to the Topic Advanced Structural Crystals)
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18 pages, 3679 KiB  
Article
UHPLC-MS Phenolic Fingerprinting, Aorta Endothelium Relaxation Effect, Antioxidant, and Enzyme Inhibition Activities of Azara dentata Ruiz & Pav Berries
by Lucia Cuesta Ramos, Javier Palacios, Ruth E. Barrientos, Jessica Gómez, Juan Manuel Castagnini, Francisco J. Barba, Alejandro Tapia, Adrián Paredes, Fredi Cifuentes and Mario J. Simirgiotis
Foods 2023, 12(3), 643; https://doi.org/10.3390/foods12030643 - 2 Feb 2023
Cited by 6 | Viewed by 3199
Abstract
Azara dentata Ruiz & Pav. is a small Chilean native plant from Patagonia, a producer of small white reddish berries. For the first time, the proximal analysis of the fruits, phenolic fingerprinting, the antioxidant activity, and the enzymatic inhibition and relaxation effects in [...] Read more.
Azara dentata Ruiz & Pav. is a small Chilean native plant from Patagonia, a producer of small white reddish berries. For the first time, the proximal analysis of the fruits, phenolic fingerprinting, the antioxidant activity, and the enzymatic inhibition and relaxation effects in rat aorta induced by the ethanolic extract of these fruits were investigated. The proximal composition and the mineral (Ca: 2434 ± 40 mg/kg; Mg: 702 ± 13 mg/kg; Fe: 117.1 ± 1.6 mg/kg; Zn: 16.1 ± 0.4 mg/kg) and heavy metal (As: 121 ± 11 µg/kg; Cd: 152 ± 5 µg/kg; Hg: 7.7 ± 1.3 µg/kg; Pb 294 ± 4 µg/kg) contents were analyzed. Anthocyanins, flavonoids, phenolic acids, and coumarins were identified using UHPLC-PDA-QTOF-MS. The ethanolic extracts showed a total phenolic content of 23.50 ± 0.93 mg GAE/g extract. In addition, the antioxidant activity was assessed using both DPPH and TEAC (28.64 ± 1.87 and 34.72 ± 2.33 mg Trolox/g of dry fruit, respectively), FRAP (25.32 ± 0.23 mg Trolox equivalent/g dry fruit), and ORAC (64.95 ± 1.23 mg Trolox equivalents/g dry fruit). The inhibition of enzymatic activities (acetylcholinesterase IC50: 2.87 + 0.23 µg extract/mL, butyrylcholinesterase IC50: 6.73 + 0.07 µg extract/mL, amylase IC50: 5.6 ± 0.0 µg extract/mL, lipase IC50: 30.8 ± 0.0 µg extract/mL, and tyrosinase IC50: 9.25 ± 0.15 µg extract/mL) was also assessed. The extract showed 50–60% relaxation in rat aorta (intact), mediated thorough the release of endothelial nitric oxide. Our results suggest that A. dentata is a good source of compounds with the capacity to inhibit important enzymes, can be hypotensive, and can thus have good potentiality as supplements in the amelioration of neurodegenerative diseases and could also have potential to be used to develop new functional foods. The study highlights the benefits of these neglected small fruits and could boost their consumption. Full article
(This article belongs to the Section Food Nutrition)
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15 pages, 3006 KiB  
Article
In Vitro Animal Model for Estimating the Time since Death with Attention to Early Postmortem Stage
by Michal Szeremeta, Paulina Samczuk, Karolina Pietrowska, Tomasz Kowalczyk, Katarzyna Przeslaw, Julia Sieminska, Adam Kretowski, Anna Niemcunowicz-Janica and Michal Ciborowski
Metabolites 2023, 13(1), 26; https://doi.org/10.3390/metabo13010026 - 23 Dec 2022
Cited by 10 | Viewed by 2716
Abstract
Estimating the postmortem interval (PMI) has remained the subject of investigations in forensic medicine for many years. Every kind of death results in changes in metabolites in body tissues and fluids due to lack of oxygen, altered circulation, enzymatic reactions, cellular degradation, and [...] Read more.
Estimating the postmortem interval (PMI) has remained the subject of investigations in forensic medicine for many years. Every kind of death results in changes in metabolites in body tissues and fluids due to lack of oxygen, altered circulation, enzymatic reactions, cellular degradation, and cessation of anabolic production of metabolites. Metabolic changes may provide markers determining the time since death, which is challenging in current analytical and observation-based methods. The study includes metabolomics analysis of blood with the use of an animal model to determine the biochemical changes following death. LC-MS is used to fingerprint postmortem porcine blood. Metabolites, significantly changing in blood after death, are selected and identified using univariate statistics. Fifty-one significant metabolites are found to help estimate the time since death in the early postmortem stage. Hypoxanthine, lactic acid, histidine, and lysophosphatidic acids are found as the most promising markers in estimating an early postmortem stage. Selected lysophosphatidylcholines are also found as significantly increased in blood with postmortal time, but their practical utility as PMI indicators can be limited due to a relatively low increasing rate. The findings demonstrate the great potential of LC-MS-based metabolomics in determining the PMI due to sudden death and provide an experimental basis for applying this attitude in investigating various mechanisms of death. As we assume, our study is also one of the first in which the porcine animal model is used to establish PMI metabolomics biomarkers. Full article
(This article belongs to the Special Issue Animal and Cellular Models in Metabolomics Research Volume 2)
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24 pages, 4050 KiB  
Article
Near-Infrared Metabolomic Fingerprinting Study of Lichen Thalli and Phycobionts in Culture: Aquaphotomics of Trebouxia lynnae Dehydration
by Irene Bruñas Gómez, Monica Casale, Eva Barreno and Myriam Catalá
Microorganisms 2022, 10(12), 2444; https://doi.org/10.3390/microorganisms10122444 - 10 Dec 2022
Cited by 7 | Viewed by 2970
Abstract
Near-infrared spectroscopy (NIRS) is an accurate, fast and safe technique whose full potential remains to be exploited. Lichens are a paradigm of symbiotic association, with extraordinary properties, such as abiotic stress tolerance and adaptation to anhydrobiosis, but subjacent mechanisms await elucidation. Our aim [...] Read more.
Near-infrared spectroscopy (NIRS) is an accurate, fast and safe technique whose full potential remains to be exploited. Lichens are a paradigm of symbiotic association, with extraordinary properties, such as abiotic stress tolerance and adaptation to anhydrobiosis, but subjacent mechanisms await elucidation. Our aim is characterizing the metabolomic NIRS fingerprints of Ramalina farinacea and Lobarina scrobiculata thalli, and of the cultured phycobionts Trebouxia lynnae and Trebouxia jamesii. Thalli collected in an air-dry state and fresh cultivated phycobionts were directly used for spectra acquisition in reflectance mode. Thalli water peaks were associated to the solvation shell (1354 nm) and sugar–water interactions (1438 nm). While northern–southern orientation related with two hydrogen bonded (S2) water, the site was related to one hydrogen bonded (S1). Water, lipids (saturated and unsaturated), and polyols/glucides contributed to the profiles of lichen thalli and microalgae. R. farinacea, with higher desiccation tolerance, shows higher S2 water than L. scrobiculata. In contrast, fresh phycobionts are dominated by free water. Whereas T. jamesii shows higher solvation water content, T. lynnae possesses more unsaturated lipids. Aquaphotomics demonstrates the involvement of strongly hydrogen bonded water conformations, polyols/glucides, and unsaturated/saturated fatty acids in the dehydration process, and supports a “rubbery” state allowing enzymatic activity during anhydrobiosis. Full article
(This article belongs to the Special Issue Feature Collection in Environmental Microbiology Section 2021-2022)
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