Search Results (1,653)

We present the cloning and heterologous expression of the α-amylase gene amyBL159 from a thermophilic strain Bacillus licheniformis MGMM159, which was isolated from wastewater sediments self-heated to 70 °C. The gene was successfully cloned into the pET22b and pHT01 vectors, expressed and AmyBL159_Ec and AmyBL159_Bs recombinant α-amylases were purified from Escherichia coli BL21(DE3)pLys and Bacillus subtilis 168 strains, respectively. The AmyBL159_Ec enzyme was most active in the range of 75–95 °C, while AmyBL159_Bs showed maximum activity at temperatures from 45 to 75 °C. AmyBL159_Bs was shown to be more thermostable. Both enzymes were active over a broad pH range of 4.0–12.0. It was shown that Mn²⁺ ions enhanced the activity of both enzymes (up to 163% for AmyBL159_Ec and 142% for AmyBL159_Bs). These results highlight the importance of choosing an expression system for modulating the functional characteristics of recombinant α-amylase. The obtained AmyBL159_Ec and AmyBL159_Bs enzymes are promising for biotechnological applications under extreme conditions. The structure of the α-amylase was generated using the AlphaFold 3 web service. A structure–function analysis of this enzyme and previously studied α-amylases from B. licheniformis identified significant amino acid substitutions at positions 134(133) and 210(209) of the amino acid chain which may contribute to enhanced enzyme thermostability. Full article

This study aimed to investigate the chemical composition, antioxidant activity, and in vitro antidiabetic properties of extracts obtained from ripe, unripe, and fermented (unripe) cornelian cherry (Cornus mas L.) fruits. Polyphenols were identified using UPLC-ESI-qTOF-MS/MS and quantified by HPLC-PDA. Antioxidant activity was evaluated using ABTS, DPPH, and FRAP assays, while enzyme inhibitory activity was determined for α-glucosidase and α-amylase. Additionally, the effects of C. mas extracts on insulin sensitivity in adipocytes were investigated. The study’s results showed that each of the extracts tested contained varying proportions of substances with proven health-promoting properties. The extract from ripe fruits was characterized by the highest loganic acid content, whereas the extract from fermented unripe fruits contained a high amount of gallic acid, released through the hydrolysis of tannins during fermentation. The extract from unripe fruits exhibited the highest tannin content and the strongest antioxidant activity. All extracts inhibited α-glucosidase and α-amylase to a similar extent and improved insulin-stimulated glucose uptake in 3T3-L1 adipocytes without affecting INSR or SLC2A4 expression. In conclusion, extracts from unripe and fermented C. mas fruits may represent promising agents for alleviating insulin resistance and preventing type 2 diabetes. Full article

This study aimed to obtain extracts concentrated in polyphenolic compounds from Sorbus aucuparia fruits and evaluate their antioxidant, antidiabetic, anti-inflammatory, and cytotoxic potential. Two modern extraction methods were used, ultrasound-assisted extraction (UAE) and accelerated solvent extraction (ASE), to obtain hydroalcoholic extracts (50% EtOH v/v, 15% mass), then the extracts were purified and concentrated by membrane technologies and analyzed spectrophotometrically and chromatographically. HPLC analysis revealed the predominant polyphenolic compounds as chlorogenic acid (526.08 ± 23.35 µg/mL), rutin (36.07 ± 1.23 µg/mL), and caffeic acid (34.41 ± 1.21 µg/mL). The antidiabetic and anti-inflammatory potential of the extracts was analyzed spectrophotometrically by testing their capacity to inhibit α-amylase and α-glucosidase, and, respectively, hyaluronidase (HYA) and lipoxygenase (LOX). The cytotoxic potential of the extracts was tested on the mouse fibroblast NCTC clone L929 cell line. The concentrated ASE extracts showed a pronounced inhibitory activity on the tested enzymes: IC₅₀α-glucosidase was 13.50 ± 0.96 µg/mL, (IC₅₀acarbose was 20.19 ± 1.67 µg/mL), IC₅₀α-amylase was 23.74 ± 1.32 µg/mL (IC₅₀acarbose was 22.65 ± 1.27 µg/mL), and IC_50LOX was 24.30 ± 1.54 µg/mL (IC₅₀ibuprofen was 26.91 ± 1.27 µg/mL), IC₅₀HYA was 43.04 ± 2.19 µg/mL (IC₅₀ibuprofen was 51.54 ± 3.67 µg/mL). Also, the concentrated UAE extracts presented inhibitory activity superior to or close to that of the standard used, as follows: IC₅₀HYA was 48.49 ± 3.15 µg/mL (IC₅₀ibuprofen was 51.54 ± 3.67 µg/mL) and IC₅₀α-glucosidase was 21.53 ± 1.25 µg/mL (IC₅₀acarbose was 20.19 ± 1.67 µg/mL). The results obtained showed that Sorbus aucuparia fruits could be used in products for diabetes and inflammatory diseases. Full article

Antarctic algae have evolved in extreme environmental conditions, developing unique metabolic adaptations with significant biotechnological potential. In this study, we explored the bioactivity of the sea-ice microalga Microglena antarctica by preparing acetone and methanol extracts from biomass cultivated at 4, 8, and 16 °C. These extracts were screened for their in vitro antioxidant properties and inhibitory activities on enzymes related to Alzheimer’s disease (acetylcholinesterase: AChE, butyrylcholinesterase: BChE), type 2 diabetes mellitus (T2DM, α-glucosidase, α-amylase), obesity (lipase), and hyperpigmentation (tyrosinase). Our screening revealed a high capacity of acetone extracts to scavenge the ABTS•⁺ radical (EC₅₀ ranging from 3.57 to 4.18 mg mL⁻¹), along with strong copper chelating activity in both acetone and methanol extracts (EC₅₀ values of 6.31 and 6.41 mg mL⁻¹). Relevant inhibition towards α-amylase (IC₅₀ values of 3.34 and 4.53 mg mL⁻¹) and tyrosinase (with IC₅₀ ranging from 3.82 to 5.47 mg mL⁻¹) was reported for acetone and methanol extracts, respectively. UHPLC-HRMS-based profiling revealed the presence of lipidic molecules, such as glycolipids, phospholipids, and betaine lipids with polyunsaturated carbon chains, together with carotenoids, including canthaxanthin and adonixanthin, which are likely responsible for the observed bioactivities. Full article

Background: Tribulus terrestris is a medicinal plant used in traditional medicine to treat certain illnesses. Though past efforts mostly focused on the fruits and roots, current research examined the phytochemical composition and bioactivity of leaf extract (LE) and seed extract (SE). Methods: GC-MS compared phytochemical profiles, and total phenolic and flavonoid content were determined. The extracts were tested for antibacterial activity (disc diffusion, MIC/MBC), antioxidant potential (DPPH, ABTS⁺), cytotoxicity (MTT assay in MCF-7 and HepG2 cells), and anti-diabetic activity (α-amylase and α-glucosidase inhibition). Expression of apoptotic genes was also investigated. Results: The LE had a superior phytochemical composition, with greater phenolic and flavonoid levels. Compared to SE, it exhibited considerably higher antibacterial activity (MIC = 6.25–25 μg/mL), antioxidant potential (IC₅₀ = 90.71–113.41 μg/mL), cytotoxicity (IC₅₀ = 105.12–126.14 μg/mL), and enzyme inhibition (IC₅₀ = 84–96.62 μg/mL). The LE also drastically reduced the expression of anti-apoptotic genes Bcl-2 and Bcl-xL in cancer cells. T. terrestris LE has significantly higher bioactive potential than SE in a range of pharmacological arenas due to its superior phytochemically complete profile. Conclusions: The findings indicate the LE as a promising candidate for the development of standardized phytotherapeutically active compounds. Full article

Foxtail millet (Setaria italica L.) is an important crop in northern China’s arid and semi-arid regions. Frequent spring droughts and limited irrigation facilities often cause poor seed germination due to insufficient soil moisture, threatening food security. The irrigation-sowing technique, which creates a localized moist microenvironment around seeds, effectively addresses this issue. However, this technique has been poorly studied, and its effects on foxtail millet seed germination remain unclear. To address this, field experiments were conducted using a two-factor split-plot design, with three drought levels and five irrigation gradients. The results showed that irrigation-sowing increased soil moisture, promoted root–shoot growth coordination, and improved germination characteristics. Transcriptome analysis of seeds under moderate drought compared the optimal irrigation treatment (13.5 m³·hm⁻²) with the non-irrigated control (0 m³·hm⁻²), identifying 2169 differentially expressed genes. Seeds receiving irrigation exhibited higher transcript abundance in pathways related to carbohydrate metabolism, energy production, secondary metabolism, and hormone signaling. Physiological measurements further showed increased α/β-amylase activity, while starch, sucrose, and cellulose content decreased. Glycolytic enzyme activity was enhanced, and ATP content increased by 125%. Additionally, phenylpropanoid metabolism was promoted, and proanthocyanidin accumulation increased by 11.5%. Hormone analysis showed that the contents of IAA and GA increased as germination progressed by 29.09% and 54.70%, respectively, while ABA content decreased. Overall, irrigation-sowing serves as an upstream moisture signal that reshapes metabolic and hormonal states associated with improved germination performance. Full article

Taraxacum mongolicum Hand.-Mazz (TMHM), a primary source of dandelion, is a globally recognized edible and medicinal plant with significant potential in food, medicine, daily chemical products, and animal husbandry. Although hypoglycemic effects have been reported in other Taraxacum species, the specific hypoglycemic constituents and mechanisms of TMHM are not well understood. The absence of comprehensive multi-target screening methodologies has hindered the elucidation of TMHM’s dual inhibitory effects on α-amylase and α-glucosidase, as well as its associated molecular mechanisms. In this study, a multi-target screening strategy was developed to concurrently evaluate α-amylase and α-glucosidase inhibition, integrating multi-target affinity ultrafiltration coupled with ultra-performance liquid chromatography-tandem mass spectrometry (MTAUF-UPLC-MS/MS), molecular docking, and molecular dynamics (MD) simulations. Using this approach, 13 dual-target inhibitors were identified from TMHM. Moreover, at least 5 of these compounds exhibited anti-diabetic activities comparable to the positive control drug acarbose, suggesting that they are principal bioactive constituents responsible for its hypoglycemic effects. Subsequent investigation of the antioxidant capacities of 7 out of the 13 bioactive compounds revealed that most exhibited more potent antioxidant activities than vitamin C (Vc). Based on these findings, molecular docking and MD simulations further validated that quercetin (8) and kaempferol (15), which demonstrated significant hypoglycemic and antioxidant activities, exhibited particularly strong affinities and stable interactions with α-amylase and α-glucosidase, respectively. In conclusion, these findings underscored the considerable potential of TMHM as a natural source of multifunctional bioactive compounds for nutraceutical, functional, and pharmaceutical applications. This study provided a critical foundation for elucidating the mechanisms underlying TMHM’s anti-diabetic effects and its therapeutic potential in mitigating diabetes-related complications, thereby facilitating future development and utilization. Full article

The Malaysian Apis cerana honey (ACH) was analysed for its physicochemical characteristics, including moisture, Baume, Brix analysis (total soluble solids and total soluble sugars), sugar profiling (fructose, glucose, sucrose, maltose, and lactose), total ash, pH, free acidity, electrical conductivity, colour analysis, and choline content. The inhibitory effects of pancreatic lipase, α-amylase, and α-glucosidase activities were also assessed. Results indicated that the sum of fructose and glucose, sucrose, and electrical conductivity were all within the recommended range following the requirements of international standards. The maximum levels were, nevertheless, exceeded by moisture, free acidity and total ash. The ACH showed potential as an anti-obesity and anti-diabetic agent by inhibiting pancreatic lipase by up to 43.4% at 0.063 mg/mL, α-amylase by up to 70% at 7.0 mg/mL and α-glucosidase by up to 67.6% at 100 mg/mL, respectively. The percentage inhibition of α-glucosidase by undiluted ACH and deionised water extract at different temperatures (4.8 ± 0.5 °C, 27 ± 0.5 °C, and 40 ± 0.5 °C) was comparable, suggesting that temperatures had little effect on the degree of inhibition. Full article

Background/Objectives: In this study, three antimicrobial peptides (1–3) were conjugated onto bare aluminum nanoparticles (NP) to produce peptide-conjugated nanoparticles (NP1–NP3) in order to evaluate their biological effects. Methods: The peptide-functionalized Al₂O₃ nanoparticles were characterized and subsequently analyzed for their antimicrobial activity against selected bacterial strains. The findings were compared with those of bare Al₂O₃ nanoparticles and free antimicrobial peptides. Through this comparison, the enhanced impact of combining nanoparticles with peptides in addressing antimicrobial resistance was demonstrated. Additionally, biofilm inhibition, microbial cell viability inhibition, DNA cleavage, antioxidant, and amylolytic activity assays were performed to comprehensively evaluate the biological functionality of the synthesized nanoparticles. Results: Although all tested samples exhibited significant antimicrobial activity, peptide-conjugated nanoparticles NP1, NP2, and NP3 provided superior activity with an MIC value of 16 mg/L. The highest biofilm inhibition activities were observed for NP2 as 53% and 70% against S. aureus and P. aeruginosa, respectively. Additionally, NP1–NP3 inhibited microbial cell viability by 100% at a concentration of 6.25 mg/L and free peptide 3 displayed E. coli inhibition as 100% at a concentration of 12.5 mg/L. Furthermore, we evaluated the biological potential of antimicrobial peptide-functionalized Al₂O₃ nanoparticles through antibiofilm, antioxidant, antidiabetic activities, and DNA cleavage assays. Peptide-conjugated nanoparticles NP1, NP2, and NP3 exhibited the highest antioxidant activities as 43.70%, 45.22%, and 59.57%, respectively. Except for NP3, the compounds were observed to act as α-amylase enzyme activators. NP and NP1–NP3 completely degraded the supercoiled circular form into small pieces. Conclusions: Our findings suggest that peptide–aluminum nanoparticle conjugation may be a promising formulation for enhancing biological activity. Further in vitro and in vivo tests may help clarify the therapeutic potential of this novel nanoformulation. Full article

The Corsican pine (Pinus nigra subsp. laricio (Poir.) Maire), a subspecies of black pine endemic to southern Italy, is widely known for the quality of its valuable timber, and the parts of the plant that are not used for this purpose are considered unusable production waste. In this study, we investigated the phytochemical profile and a series of biological activities of extracts from the female and male pine cones. The extracts were prepared by maceration with ethanol and subsequently fractionated using liquid-liquid separation. The total phenolic and flavonoid content, antioxidant potential (DPPH and β-carotene bleaching tests), anti-inflammatory activity (nitric oxide inhibition in RAW 264.7 cells), and enzymatic inhibition against pancreatic lipase and α-amylase were determined. The female cones showed a higher crude extract yield and total phenolic content (76.4 mg GAE/g) than the male cones, while the latter were richer in flavonoids. The extracts from the female cones showed higher antioxidant and pancreatic lipase inhibitory activities. On the contrary, extracts from male cones showed greater activity against α-amylase, with the dichloromethane fraction proving to be the most potent (IC₅₀ = 35.28 ± 3.08 µg/mL). The hexane fraction of female cones also showed significant anti-inflammatory activity (IC₅₀ = 107.50 ± 15.22 µg/mL). Our results reveal that the pine cones of Pinus nigra subsp. laricio (Poir.) Maire are a rich source of bioactive compounds. These results provide the first scientific evidence of the potential of extracts from this still poorly studied part of the plant for further investigation of their antioxidant and anti-inflammatory capabilities. Full article

The escalating global prevalence of obesity underscores the need for effective and sustainable nutritional interventions. Functional foods, especially white kidney beans (Phaseolus vulgaris), show a promising avenue to link fundamental biochemical insights with clinically feasible interventions, supporting their potential as an adjunct dietary strategy for managing and preventing obesity. This review critically examines the mechanistic roles of white kidney bean in weight regulation, which includes suppression of starch digestion, attenuation of postprandial glycemia, modulation of appetite and satiety, and hypolipidemic effects. Clinical and preclinical evidence supports the potential of white kidney bean as a nutraceutical for metabolic health, demonstrating consistent reductions in body fat mass, glycemic excursion, and overall weight. Nevertheless, significant limitations persist, including heterogeneity in trial designs, absence of dose standardization, and inadequate long-term safety assessments. Furthermore, this review addresses food fortification, advancements in supplement formulation, and cooking techniques that enhance both consumer acceptability and the bioactivity of white kidney bean (WKB), along with the significance of regulatory standards to ensure safety and quality. Future research should integrate clinical, molecular and food technology methods to improve the translation of experimental findings into precision nutritional strategies for obesity management. Full article

Digestive enzymes play a crucial role in carbohydrate hydrolysis and subsequent glucose absorption, and their inhibition can contribute to improved glycemic regulation. Legumes, with their inherent enzyme-inhibitory properties, offer a natural approach for achieving this. In this study, accessions of M. uniflorum (Lam.) Verdc, an underutilized legume, were evaluated in vitro for their α-amylase and α-glucosidase inhibitory activities, as well as their protein, amylose, and resistant starch contents. The results revealed significant variation among the accessions. PI 174827 01 SD (IC₅₀ = 23.29 ± 0.01 µg/mL) and PI 173901 01 SD (IC₅₀ = 24.60 ± 0.01 µg/mL) demonstrated strong inhibition of α-amylase and α-glucosidase, respectively. Protein content ranged from 13.81 to 27.08%w/w d.w., with PI 180437 01 SD showing the highest percentage. Total starch content ranged from 27.48 to 54.70%w/w d.w., amylose from 27.05 to 48.13%w/w d.w., and resistant starch from 5.89% to 7.09%w/w d.w., with PI 174827 01 SD exhibiting both higher amylose and resistant starch contents. These findings suggest that M. uniflorum accessions possess enzyme-inhibitory and nutritional components that could be harnessed to develop functional foods, nutraceuticals, and pharmaceuticals for the management of diabetes and obesity. Full article

To clarify the effects of hydrogen peroxide (H₂O₂) seed soaking on the germination and stress resistance of different edible bean seeds, seeds of mung bean (Vigna radiata L. ‘Keda Green No. 2’), cowpea (Vigna unguiculata L. ‘Keda Cowpea No. 1’), and red bean (Vigna umbellata Thunb. ‘Jihong 352’) were soaked in a 50 mmol/L H₂O₂ solution. The study examined the germination and growth-related physiological indices of seeds after soaking. The results showed that hydrogen-peroxide-primed seeds of mung bean (GBH), cowpea (CBH), and red bean (RBH) exhibited significant improvements in germination performance and physiological activity compared with their respective controls (GBCK, CBCK, and RBCK). The results indicated that H₂O₂ soaking significantly improved the germination ability of the seeds, with the germination rate of mung beans, cowpeas, and red beans increasing by 48.89%, 21.11%, and 18.89%, respectively, and the germination percentage increasing by 31.11%, 24.45%, and 17.77%. Additionally, H₂O₂ soaking enhanced the activity of α-amylase, protease, and the antioxidant enzymes peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT); increased the soluble sugar and soluble protein content in the seeds; and reduced the malondialdehyde (MDA) content. The experiment demonstrated that H₂O₂ promotes the germination of mung bean, cowpea, and red bean seeds by influencing antioxidant enzyme activity, the breakdown of storage substances, and the regulation of germination-related substances, thereby improving seedling adaptation to environmental stress. This study aims to improve the germination rate of legume seeds using H₂O₂ treatment, providing a theoretical basis for techniques to enhance seed vigor, especially for seeds that perform poorly in germination under normal conditions. Full article

Background: Alginate is a naturally occurring anionic polysaccharide extracted from brown marine algae and widely explored for biomedical applications due to its biocompatibility and functional versatility. This study aims to extract and compare alginates from two Red Sea brown algae, Turbinaria ornata (TA) and Hormophysa cuneiformis (HA), and to evaluate how structural differences influence their therapeutic properties. Methods: Alginate was isolated by sequential acid–alkaline extraction and characterized using FTIR, XRD, TGA, elemental analysis, and HPLC. Biological activities were assessed through antioxidant, anti-inflammatory, antidiabetic, neuroprotective, and hepatoprotective assays, supported by molecular docking and gene ontology interaction analysis. Results: Distinct physicochemical variations were observed between HA and TA. TA exhibited stronger antioxidant (IC₅₀ = 25.89 µg/mL), anti-inflammatory (COX-1 IC₅₀ = 69.61 µg/mL), antidiabetic (α-amylase IC₅₀ = 45.14 µg/mL), and hepatoprotective activities (IC₅₀ = 118.21 µg/mL), whereas HA displayed superior neuroprotective potential through butyrylcholinesterase inhibition (IC₅₀ = 39.01 µg/mL). Molecular docking supported the in vitro findings by confirming interactions with key protein targets associated with oxidative stress and metabolic pathways. Conclusions: Structural variation between species-derived alginates directly impacts their biological activities. TA represents a promising candidate for metabolic and anti-inflammatory therapies, while HA may be more suitable for neuroprotective interventions. These results emphasize the importance of source-specific alginate selection for developing targeted pharmaceutical applications. Full article

This study characterized leaf extracts of Cymbopogon flexuosus (Ryukyu Lemongrass Corporation, Okinawa, Japan) and evaluated the bioaccessibility and bioactivities of phenolic compounds following a simulated in vitro gastrointestinal model of digestion (in vitro GID) of plant material. Undigested (controls, AqC, EtC) and digested aqueous (AqD) and ethanolic (EtD) extracts were analyzed. Control extracts contained higher total phenolics and flavonoids than digested ones, with EtC showing the highest values. UHPLC-QToF-MS (ultra-high-performance liquid chromatography system coupled to a quadrupole time-of-flight mass spectrometer) identified 32 compounds, including phenolic acids, flavone aglycones, C-glycosides, and derivatives. Hydroxybenzoic acids, coumaric acid, caffeic esters, flavones, tricin derivatives, vitexin, and isoorientin exhibited reduced recovery, while coumaric acid hexoside, ferulic acid hexoside, and isoschaftoside/schaftoside exceeded 100% recovery, suggesting release from the matrix. Some compounds were absent from AqD, and many were found in the pellet, indicating potential colonic metabolism. Antioxidant activity (DPPH, reducing power, β-carotene/linoleic acid) was stronger in controls but always weaker than BHT/ascorbic acid. Extracts mildly inhibited α-amylase but more strongly inhibited α-glucosidase as shown with applied enzyme inhibition assays, especially EtD (76.93% at a concentration of 10 mg/mL), which showed stronger activity than controls but remained below acarbose (87.74% at 1 mg/mL). All extracts promoted HaCaT keratinocyte growth and reduced HCT-116 colon cancer cell viability at 250 µg/mL, with the strongest effects in AqC and AqD. Overall, GID decreased antioxidant activity but enhanced antidiabetic potential, confirming the safety and selective anticancer effects of C. flexuosus extracts. Full article