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Biocatalytic membranes have great potential in various industrial sectors, with the immobilization of enzymes being a crucial stage. Immobilizing enzymes through covalent bonds is a complex and time-consuming process for large-scale applications. Polydopamine (PDA) offers a more sustainable and eco-friendly alternative for enzyme immobilization. Therefore, surface modification with polydopamine as mussel-inspired antifouling coatings has increased resistance to fouling. In this study, α-amylase enzyme was covalently bound to a bioactive PDA-coated polyethersulfone (PES) membrane surface using cyanuric chloride as a linker. The optimal activity of α-amylase enzyme immobilized on PES/PDA membrane was obtained at temperature and pH of 55°C and 6.5, respectively. The immobilized enzyme can be reused up to five reaction cycles with 55 % retention of initial activity. Besides, it maintained 60 % of its activity after being stored for five weeks at 4°C. Additionally, the immobilized enzyme demonstrated increased Michaelis constant and maximum velocity values during starch hydrolysis. The results of the biofouling experiment of various membranes in a dead-end cell demonstrated that the PES membrane's water flux increased from 6722.7 Lmh to 7560.2 Lmh after PDA modification. Although α-amylase immobilization reduced the flux to 7458.5 Lmh due to enhanced hydrophilicity, compared to unmodified membrane. The findings of this study demonstrated that the membrane produced through co-deposition exhibited superior hydrophilicity, enhanced coating stability, and strong antifouling properties, positioning it as a promising candidate for industrial applications. [Display omitted] • α-Amylase was covalently immobilized onto polydopamine-functionalized polyethersulfone membrane. • Surface modification with polydopamine generated a mussel-inspired coating to inhibit fouling. • The immobilization technique effectively preserved the activity of α-amylase enzyme. • The biocatalyst membrane exhibited superior stability and strong antifouling properties. [ABSTRACT FROM AUTHOR]

Diabetes is marked by postprandial hyperglycemia (PHG), an abnormal rise in blood glucose after meals. A key therapeutic goal to reduce PHG is the inhibition of α‐amylase (αAM) and α‐glucosidase (αGL), enzymes that break down carbohydrates into sugars. Cucurbita moschata has been shown to inhibit both enzymes. However, its inhibition mechanism has not been explored. This study investigated the in vitro inhibition mechanisms of αAM and αGL and conducted a metabolomic analysis of C. moschata (edible part) water‐extract (CME), aiming to preliminarily identify its bioactive compounds (BCs). The inhibitory mechanisms were determined using Lineweaver–Burk plots. The BCs were identified and quantified using HPLC‐QTOF‐MS, employing both targeted and untargeted metabolomic approaches. CME had a significant higher effect (p < 0.05) on αAM activity than against αGL with IC50 of 28.99 and 698.42 mg/mL, respectively. The extract showed mixed and uncompetitive type inhibitions on αAM and αGL, respectively. The lowest inhibition constant (Ki) was 47.68 mg/mL on αAM activity at 20 mg/mL. Untargeted metabolic profiling by UPLC‐MS‐ESI‐QTOF putatively identified 30 compounds in CME, such as amino acids, vitamins, phytohormones, fatty acids, cucurbitacins and phenolic acids, and flavonoids. Functional analysis of CME identified significant pathways, including pantothenate and CoA biosynthesis and phenylpropanoids, among others. The targeted analysis by UPLC‐MS‐ESI‐QqQ allowed us to identify 12 compounds, with l‐phenylalanine, p‐hydroxybenzoic, and p‐coumaric acid as majors. This study demonstrated the inhibitory potential of CME on αAM and αGL activities, which may be attributed to its metabolites. Thus, this plant represents a valuable source of BC against PHG.Practical Application: The research highlights that Cucurbita moschata has significant potential in managing postprandial hyperglycemia in diabetic patients by inhibiting enzymes like α‐amylase and α‐glucosidase. In addition, the identification of its compounds emphasizes its importance as a source of bioactive compounds. Therefore, C. moschata could be effectively utilized in the development of nutraceuticals or as an ingredient in functional foods specifically designed for postprandial hyperglycemia management. Thus, integrating C. moschata as part of the daily diet could offer patients with diabetes a natural alternative to control their blood glucose levels after eating. [ABSTRACT FROM AUTHOR]

Simple Summary: Ruminal protein degradation of seven feedstuffs was estimated in sacco and in vitro by using a protease assay. In sacco protein degradation data were used as reference. The accuracy of the protease assay is reduced by two methodological aspects. During in vitro incubation, microbial activity induced by microbial colonization of the feedstuff may increase, requiring antibiotic solutions in protease assays. The protease alone cannot sufficiently hydrolyze protein–carbohydrate complexes. Therefore, a carbohydrase of fiber- or starch-hydrolyzing activity was added to the protease assay as a simultaneous incubation. The antibiotic solution reduced protein degradation by protease. The antibiotic solution is recommended to prevent microbial activity and improve standardization of the protease assay. Differences between in sacco and in vitro degradation data were not essentially reduced by additional carbohydrases. Unfavorable incubation conditions and the inhibitory effect of protease on the carbohydrase activity during simultaneous incubation may be responsible for the insufficient hydrolysis of protein–carbohydrate complexes by the carbohydrases. It does not seem promising to incubate protease and carbohydrase simultaneously. The objectives of the study were to examine the effect of an antibiotic solution applied in the Streptomyces griseus protease method (SGPM) and the effect of carbohydrases in SGPM on the effective crude protein (CP) degradation (ED) with reference to in sacco ED. For this purpose, the ruminal CP degradation of rapeseed meal, dried distillers' grains with solubles, wheat grain, corn grain, corn silage, grass silage and partial crop field pea silage was determined in sacco using three rumen-fistulated dairy cows and in vitro using SGPM. The impact of the antibiotic solution on CP degradation by S. griseus protease was investigated by supplementing SGPM with Penicillin–Streptomycin solution to reduce microbial mass proliferation during incubation. The carbohydrase α-amylase or Viscozym® L (cell wall-degrading enzyme mixture) was added to the SGPM at four different doses simultaneously as a co-incubation to improve feed protein accessibility. For most feedstuffs, ED was lower when the antibiotic solution was used in SGPM (p < 0.05). The use of an antibiotic solution in the SGPM is recommended to standardize the SGPM. The in sacco ED values were significantly underestimated by the SGPM and by the SGPM with co-incubated carbohydrase (p < 0.05). Co-incubation of S. griseus protease and carbohydrase was not successful in reducing the differences to the in sacco CP degradation. [ABSTRACT FROM AUTHOR]

The cloud forest (CF), a hugely biodiverse ecosystem, is a hotspot of unexplored plants with potential for discovering pharmacologically active compounds. Without sufficient ethnopharmacological information, developing strategies for rationally selecting plants for experimental studies is crucial. With this goal, a CF metabolites library was created, and a ligand-based virtual screening was conducted to identify molecules with potential hypoglycemic activity. From the most promising botanical families, plants were collected, methanolic extracts were prepared, and hypoglycemic activity was evaluated through in vitro enzyme inhibition assays on α-amylase, α-glucosidase, and dipeptidyl peptidase IV (DPP-IV). Metabolomic analyses were performed to identify the dominant metabolites in the species with the best inhibitory activity profile, and their affinity for the molecular targets was evaluated using ensemble molecular docking. This strategy led to the identification of twelve plants (in four botanical families) with hypoglycemic activity. Sida rhombifolia (Malvaceae) stood out for its DPP-IV selective inhibition versus S. glabra. A comparison of chemical profiles led to the annotation of twenty-seven metabolites over-accumulated in S. rhombifolia compared to S. glabra, among which acanthoside D and cis-tiliroside were noteworthy for their potential selective inhibition due to their specific intermolecular interactions with relevant amino acids of DPP-IV. The workflow used in this study presents a novel targeting strategy for identifying novel bioactive natural sources, which can complement the conventional selection criteria used in Natural Product Chemistry. [ABSTRACT FROM AUTHOR]

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Type-2 diabetes affects an increasing percentage of the world's population and its control through dietary management, involving the consumption of health-promoting foods or their derived supplements, is a common strategy. Several mushroom species have been demonstrated to be endowed with antidiabetic properties, resulting from their ability in improving insulin sensitivity and production, or inhibiting the carbohydrate-hydrolyzing enzymes α-amylase and α-glucosidase. This study aimed to investigate for the first time the hypoglycemic properties of the edible mushroom Leccinum scabrum (Bull.) Gray. Mushroom extracts were prepared through the microwave-assisted extraction (MAE) technique using green solvents with different polarity degrees. The inhibition activity of all the obtained extracts on both α-glucosidase and α-amylase was evaluated and the highest activity was observed for the EtOAc extract which showed an IC50 value about 60-fold lower than the reference compound 1-deoxynojirimycin (DNJ) on α-glucosidase (0.42 ± 0.02 and 25.4 ± 0.6 µg/mL, respectively). As expected on the basis of the literature data concerning both α-glucosidase and α-amylase inhibition, a milder inhibition activity on pancreatic α-amylase was observed. Preliminary in vivo tests on Drosophila melanogaster carried out on the most active obtained extract (EtOAc) confirmed the in vitro observed hypoglycemic activity. Finally, the EtOAc extract metabolic profile was determined through GC-MS and HRMS analyses. [ABSTRACT FROM AUTHOR]

In this study, we developed a sensitive method for monitoring α‐amylase using a fluorogenic approach based on the host‐guest complexation between an amphiphilic pyrenyl derivative (1) and γ‐cyclodextrins (γ‐CDs). The compound 1 self‐assembles into nanofibrils in aqueous solutions. Upon the introduction of γ‐CD, compound 1 forms an inclusion complex with it. This complex then participates in the formation of a 2 : 2 complex with another complex, leading to strong excimer fluorescence. Upon interaction with α‐amylase, γ‐CD undergoes hydrolysis, leading to the regeneration of nanofibrils, which is accompanied by a decrease in excimer fluorescence and an increase in monomeric fluorescence. This ratiometric fluorescence color change enables the sensitive detection of low levels of α‐amylase in human urine, offering a practical approach for early screening of pancreatic‐related diseases. [ABSTRACT FROM AUTHOR]

The carbazole-3-carbaldehyde 2, produced by N-ethyl carbazole via Vilsmeier-Haack reaction, was subjected to Dakin type oxidation with H2O2 and H2SO4 in methanol to produce the carbazole-3-ol 3. The reaction of 3 with a range of commercially available α-haloketones 4a–f in the presence of Al2O3 as catalyst in xylene led to their regio-selective cyclization to afford the furo[2,3-c]carbazoles 5a–f. Identification of the furo[2,3-c]carbazoles 5a–f were performed through 1H NMR,13C NMR, FT-IR and high resolution mass spectrometry. Single crystal X-ray diffraction analysis was employed to further confirm the structures of the some of the targeted compounds. In vitro antidiabetic activities of the newly synthesized furocarbazoles 5a–e were investigated utilizing α-glucosidase and α-amylase enzymes. The biological evaluation revealed the obvious efficiencies of the targeted molecules toward the α-glucosidase enzyme inhibition with the potent IC50 values compared to the standard acarbose. In the case of α-glucosidase inhibition, the furo[2,3-c]carbazoles chloro substituted 5c and nitro substituted 5f were found to be more potent than acarbose with the values of 215.0 and 162.70 μM, respectively. On the other hand, the compound 5f was found to be only promising candidate for α-amylase enzyme but not as effective as the standard acarbose. [ABSTRACT FROM AUTHOR]

• Mass multiplication of Mundulea sericea's callus. • To investigate the characterisation of bioactive compounds from the callus of M. sericea through GC–MS. • Detection of anti-diabetic agents through in silico. Mundulea sericea , a rare plant of the fabaceae family, has high medicinal value but a low regeneration rate in its natural habitat. The present study aims to conduct in vitro propagation and screen the potential bioactive compounds in the callus through in silico analysis. The explants were collected, sterilized, and inoculated in a standardized MS media with growth regulators in vitro to obtain sterile plants. The experiment resulted in the formation of a greenish-brown callus at a rate of 82.41 % on MS medium supplemented with BAP (6.66 µM L−1). The callus was powdered and subjected to GC–MS analysis. A total of 31 compounds were identified in the methanolic extract, along with their corresponding peak values. The compounds underwent ADME analysis for toxicity predictions. Eight compounds were chosen further based on pharmacokinetic and pharmacodynamic properties. The selected ligands were docked with anti-diabetic targets α-amylase and α-glucosidase. As a result, the compound CID 5283663 exhibited a strong binding affinity of −9.3 with the α-amylase receptor. The ligand CID 33010 exhibits a strong interaction of −7.5 with the α-glucosidase receptor. Glibenclamide and acarbose were used as standards. This study is the first attempt to identify compounds in the callus extract that could inhibit α-amylase and α-glucosidase enzymes. The findings from molecular docking analysis suggest these compounds could become potent treatments for diabetics, showcasing the promise of natural sources in drug discovery. [ABSTRACT FROM AUTHOR]

An accurate characterization of the fine molecular structure of starch in terms of the chain length distribution (CLD) of amylopectin requires complete dispersion of the starch in a solvent, either buffer or water and the non-interference of this on the subsequent enzymatic debranching of starch with isoamylase (pretreatments before determining the CLD). In this study, the HYLON V and HYLON VII starches (commercial high-amylose corn starches) were selected, since high-amylose starches typically present difficulties in their dispersion and, therefore, the study of their fine structure can commonly present errors. The goal of this research was to study two water dispersion treatments (1. boiling water for 1 h and 2. autoclaving at 120 °C and 15 Psi for 15 min) to achieve a fast and reliable method of the fine structure of two high-amylose corn starches using high-performance size exclusion chromatography coupled with a refractive index detector (HPSEC-RID) and high-performance anion-exchange chromatography coupled a pulsed amperometric detector (HPAEC-PAD). The HPSEC-RID chromatograms of both starches showed better resolution using autoclaving than boiling water treatment. The two starches tested had higher amylose and degree of polymerization (DP) values utilizing the autoclaving than the boiling water treatment. However, no appreciable differences were observed in the CLD values employing HPAEC-PAD for either of the two water dispersion treatments. Lastly, the water dispersion treatment by autoclaving before enzymatic debranching is recommended to obtain a more accurate quantification of the amylose content of high-amylose starches. [ABSTRACT FROM AUTHOR]

Two previously undescribed dihydrochalcones, 4,2′-dihydroxy- [2″, 2″-dimethylpyrano (3″, 4″: 4′, 5′]) dihydrochalcone (1) and 4, 2′, 4′, 2″-tetrahydroxy-5′-(3″-methylbut-3″-enyl) dihydrochalcone (2), along with six known compounds (3 - 8) were isolated from chloroform-methanol (1:1) extract of Baphia Massaiensis twigs. Their chemical structures were determined using NMR analysis, mass spectrometry, electronic circular dichroism (ECD), density functional theory (DFT) calculations, and a thorough comparison with reported data from the literature. All the isolated compounds (1−8) were assessed for their antioxidant activities, α-amylase and α-glucosidase inhibition effects. Dihydrochalcone 1 showed significant antioxidant activity with IC 50 value of 118.8 ± 2.95 µM, and a more potent antioxidant activity was exhibited by compounds 3 and 6 with IC 50 values of 84.3 ± 0.07 µM and 62.1 ± 1.59 µM, respectively, compared to the positive control, gallic acid with IC 50 value of 92.9 µM. Dihydrochalcone 2 showed moderate inhibition against α-amylase and α-glucosidase with IC 50 values of 44.0 ± 4.83 µM and 51.7 ± 9.72 µM compared to the standard acarbose with IC 50 values of 22.9 ± 0.92 µM and 31.5 ± 0.63 µM, respectively. Molecular docking studies of the new dihydrochalcones 1 and 2 , revealed the active binding sites which support the observed α-amylase and α-glucosidase inhibitory effects. [Display omitted] • Two undescribed dihydrochalcones were isolated from Baphia massaiensis. • The structures were confirmed by extensive spectroscopic analyses. • Compounds 1 and 2 showed significant inhibitory activities against α-amylase and α-glucosidase. • Compounds 1 , 3 and 6 also showed potent antioxidant activities. [ABSTRACT FROM AUTHOR]

Seeking food-derived antioxidants and inhibitors of α-glucosidase and α-amylase has been recognized as an effective way for managing diabetes. Coffee silver skin (CSS) is rich in phenolic compounds, which may be potential agents as antioxidants and for α-glucosidase and α-amylase inhibition. But whether phenolics in different forms show similar bioactivity remains unknown. In this study, phenolic compounds in CSS were extracted as free phenolics (FPs), esterified phenolics (EPs), and bound phenolics (BPs). The phenolic profiles and antioxidant activities of them were investigated. Their inhibitory effects on α-glucosidase and α-amylase were analyzed, and the inhibitory mechanisms were elucidated by molecular docking and molecular dynamic simulation. Results showed that FPs exhibited the best antioxidant ability and inhibitory effects on α-glucosidase and α-amylase. A total of 17 compounds were identified in FPs with 3-caffeoylquinic acid, 4-feruloylquinic acid, and dicaffeoylquinic acids as the dominant ones. Typical phenolics in FPs could bind to α-glucosidase and α-amylase through hydrogen bonds and form hydrophobic interaction with several key amino acid residues. In addition, 3,4-dicaffeoylquinic acid and 3-caffeoylquinic acid might be the principal components that account for the inhibitory effect of FPs on α-glucosidase. The results of this study may provide some scientific support for CSS utilization as a health-beneficial component in functional food development for type 2 diabetes mellitus management. [ABSTRACT FROM AUTHOR]

We investigated the hypoglycemic activity and pharmacokinetic study of two synthesized benzoyl benzodioxol derivatives, compound I (methyl 2-(6-(2-bromobenzoyl)benzo[d][1,3]dioxol-5-yl)acetate), and compound II, 2-(6-benzoylbenzo[d][1,3]dioxol-5-yl)acetic acid, which showed very strong α-amylase inhibiting activity in our previous study. Then, diabetes was induced by the injection of streptozotocin to mice. The molecular docking simulations and analyses of density functional theory analyses were conducted to study the binding interactions with human pancreatic alpha-amylase, and their pharmacokinetic properties were further evaluated by ADMET profiling. Compound I showed the most important hypoglycemic effect, decreasing the blood glucose by 32.4%, higher than that of compound II by 14.8% and even the positive control acarbose by 22.9%. Histopathological examination revealed that diabetic livers showed portal inflammation with some apoptotic hepatocytes due to streptozotocin treatment, whereas controls without any treatment maintained normal liver architecture. Molecular docking studies gave results for the best binding affinity of the compound I, through its strong water bridges and π–π interactions, and also through analysis with density functional theory, was more stable and reactive when compared to compound II. Further ADMET analysis showed that both compounds shared a promising pharmacokinetic profile, and compound I had the potential for CNS penetration. Thus, compound I was selected as the best candidate for developing new hypoglycemic agents with potent efficacy, good binding interactions, and excellent pharmacokinetic properties. [ABSTRACT FROM AUTHOR]

Diabetes is a major health concern and is approaching epidemic proportions worldwide. In 2021, diabetes mellitus was responsible for 6.7 million deaths across the globe. Mortality due to diabetes is predicted to rise nearly 10‐fold by 2030 and 783 million by 2045. Wheat starch, which constitutes about 70% of the endosperm, is a key component of wheat grain. The rapid hydrolysis of wheat starch can result in elevated postprandial glucose levels, leading to diabetes. The increase in blood glucose levels is primarily due to carbohydrate hydrolysis, catalyzed by the enzymes α‐amylase and α‐glucosidase. Although various medications are available for treating diabetes, most of them are costly and may lead to adverse effects. Natural herbs like fenugreek are recommended in traditional medicine for regulating blood glucose levels. This investigation aimed to study the effect of fenugreek seed extract (FSE) on in vitro starch hydrolysis by pancreatic α‐amylase and the ultrastructure of starch. Wheat cultivars were characterized for their total starch, amylose content, and resistant starch content, and were screened for their predicted glycemic index. Microscopic studies were conducted to analyze the size and shape of starch granules and to compare native starch with starch treated with FSE. Significant inhibition of enzymatic starch hydrolysis was observed with FSE, with the maximum inhibitory effect caused by 0.2% FSE. These findings suggest that fenugreek could play a role in controlling blood glucose levels by reducing wheat starch hydrolysis and could be effective in managing diabetes. [ABSTRACT FROM AUTHOR]

Sikkim in North East India, has a rich ethnomedicinal practice and a large proportion of the local people rely on medicinal plants for the management of diabetes. Anti-hyperglycaemic effect of ten medicinal plants traditionally used by the local population in Sikkim to treat diabetes was evaluated using two in vitro methods - glucose uptake in yeast cells and α-amylase inhibition assay. Most of these medicinal plants demonstrated anti-hyperglycaemic effect, amongst which the leaves of Catharanthus roseus and Nyctanthes arbor-tristis displayed considerably greater activity than the other plant extracts. Both C. roseus and N. arbor-tristis showed greatest increase in glucose uptake at 25 mM glucose concentration (80.20% ± 3.01 and 78.83% ± 2.66 respectively), and maximum α-amylase inhibition of 72.29% ± 0.58 (IC50 = 154.30 μg/ml) and 69.98% ± 1.16 (IC 50 = 150.48 μg/ml) respectively at 250 μg/ml concentration of the extracts. The values were comparable to the standard drugs. Ethnomedicinal practices largely rely on observations, experience and regional beliefs. Support from scientific assessment studies will not only encourage traditional practices in the region, but also contribute in the development of alternative drugs of natural origin for diabetes. [ABSTRACT FROM AUTHOR]

During the domestication of crops, seed dormancy has been reduced or eliminated to encourage faster and more consistent germination. This alteration makes cultivated crops particularly vulnerable to pre-harvest sprouting, which occurs when mature crops are subjected to adverse environmental conditions, such as excessive rainfall or high humidity. Consequently, some seeds may bypass the normal dormancy period and begin to germinate while still attached to the mother plant before harvest. Grains affected by pre-harvest sprouting are characterized by increased levels of α-amylase activity, resulting in poor processing quality and immediate grain downgrading. In the agriculture industry, pre-harvest sprouting causes annual economic losses exceeding USD 1 billion worldwide. This premature germination is influenced by a complex interplay of genetic, biochemical, and molecular factors closely linked to environmental conditions like rainfall. However, the exact mechanism behind this process is still unclear. Unlike pre-harvest sprouting, vivipary refers to the germination process and the activation of α-amylase during the soft dough stage, when the grains are still immature. Mature seeds with reduced levels of ABA or impaired ABA signaling (weak dormancy) are more susceptible to pre-harvest sprouting. While high seed dormancy can enhance resistance to pre-harvest sprouting, it can lead to undesirable outcomes for most crops, such as non-uniform seedling establishment after sowing. Thus, resistance to pre-harvest sprouting is crucial to ensuring productivity and sustainability and is an agronomically important trait affecting yield and grain quality. On the other hand, seed color is linked to sprouting resistance; however, the genetic relationship between both characteristics remains unresolved. The identification of mitogen-activated protein kinase kinase-3 (MKK3) as the gene responsible for pre-harvest sprouting-1 (Phs-1) represents a significant advancement in our understanding of how sprouting in wheat is controlled at the molecular and genetic levels. In seed maturation, Viviparous-1 (Vp-1) plays a crucial role in managing pre-harvest sprouting by regulating seed maturation and inhibiting germination through the suppression of α-amylase and proteases. Vp-1 is a key player in ABA signaling and is essential for the activation of the seed maturation program. Mutants of Vp-1 exhibit an unpigmented aleurone cell layer and exhibit precocious germination due to decreased sensitivity to ABA. Recent research has also revealed that TaSRO-1 interacts with TaVp-1, contributing to the regulation of seed dormancy and resistance to pre-harvest sprouting in wheat. The goal of this review is to emphasize the latest research on pre-harvest sprouting in crops and to suggest possible directions for future studies. [ABSTRACT FROM AUTHOR]

This study involved the synthesis, characterization, and assessment of fourteen pyridazine analogs (designated as 1–14) to investigate their efficacy in inhibiting the α‐amylase enzyme for potential diabetes treatment using an in vitro approach. Additionally, in silico molecular docking and molecular dynamic (MD) simulations were conducted to assess the inhibitory properties of these analogs. Physicochemical and pharmacokinetic properties of the fourteen pyridazine analogs were predicted using the DataWarrior tool. Results indicated that all tested compounds demonstrated significant α‐amylase inhibitory activity, with IC50 values ranging from 81.28±0.00 to 1623.54±2.67 μM compared to the standard drug acarbose (IC50=220.42±36.40 μM). Notably, compounds 8 and 12 exhibited the most potent α‐amylase inhibitory activity, with IC50 values of 81.28±0.00 μM and 200.60±34.65 μM, respectively. Molecular docking analysis revealed binding energies ranging from −7.53 to −5.77 kcal/mol and inhibition constants ranging from 3.00 to 58.96 μM, with compounds 9, 7, 8, 5, and 3 demonstrating the best binding energies. Subsequent MD simulation analyses indicated that all five compound formed stable complexes after 100 ns MD simulation. Consequently, these compounds hold promise as potential α‐amylase inhibitors pending successful clinical validation. [ABSTRACT FROM AUTHOR]

Tungrymbai is an ethnic fermented soy food that is popular amongst the Meghalaya's Khasi and Garo tribes. In our study, we tried to standardise the traditional fermented soy product with well-characterised Lactobacillus cultures and studied its bio-functionalities. Tungrymbai, a fermented soy product, was prepared and studied for up to eight days for its biofunctional properties, total phenolic contents, isoflavone bioconversion, and anti-inflammatory activity on cell culture study during the storage study. Two lactic acid bacteria, namely K4E (Lacticaseibacillus rhamnosus KX950834.1) and K14 (Lactobacillus helveticus KU644578.1), were used for the preparation of the Tungrymbai. The storage study of Tungrymbai was carried out for eight days and was sampled every two days for further analysis. The sensory scores were greater than six up to day 6, whereas beyond that point, the scores began to decline. The pH of the Tungrymbai decreased to 5.34 with an increase in storage time, whereas the acidity increased to 0.054% lactic acid. The LAB counts were found to be highest during the initial time of the study (7.83±0.33 log CFU/mL). The maximum level of ACE inhibitory activity was noted during the initial day of the study to be 56.70±1.35%. Anti-diabetic activities such as α-glucosidase, α-amylase, and anti-lipase activities were performed for Tungrymbai. The greatest level of α-amylase inhibition was 34.07±2.31% observed during the initial time of the study, whereas α-glucosidase inhibitory activity was found highest on the eighth day of the study to be 37.67±1.24%, and lipase inhibitory activity increased with an increase in storage time up to the sixth day of the study (22.23±0.18%), which significantly decreased on the eighth day of the study. Antioxidative activity was found highest on the sixth day of the study, i.e., 73.65±0.54%. The total phenolic content of Tungrymbai decreased with an increase in storage time and was found to be highest on the initial day at 2.08±0.031mg/mL. Aglycone isoflavones, i.e., Daidzein and Genistein were estimated for Tungrymbai, where Diadzein content decreased with an increase in storage time, while Genistein content increased up to the fourth day of the study as 38.16±4.84%, which was significantly decreased on the sixth and eighth days of the study, respectively. MTT tests revealed that the lowered dosage of Tungrymbai exhibited no cytotoxicity in Raw 264.7 macrophages cell line. [ABSTRACT FROM AUTHOR]

In reaction to the expanding predominance of diabetes mellitus, curcumin nanoparticles stacked on carboxymethyl cellulose (CMC) composite were effectively synthesized, characterized, and examined utilizing UV/Vis and FTIR spectroscopy combined with transmission electron microscopy (TEM). The bioactivity of curcumin (Cur), carboxymethyl cellulose (CMC), and curcumin nanoparticles stacked with carboxymethyl cellulose (CUR-CMC) was tried through atomic docking approval as an α-amylase and α-glucosidase inhibitor. The conclusion illustrated that the curcumin-supported CMC is more potent than CUR itself self the validation presented is compared with acarbose as a reference molecule and then CUR-CMC can presented as promising in curing hyperglycemia by decreasing the absorption of glucose. [ABSTRACT FROM AUTHOR]

Pistacia species (P. spp) have been used as a treatment for various diseases, including diabetes and inflammation. This study aimed to identify the main components of flavonoids in Pistacia species and evaluate the effect of aqueous extracts of P. spp on pancreatic enzymes and on cancer cells associated with obesity in colon and rectum. HPLC was used to identify the major components of flavonoids. The potent inhibitory effect of Pistacia species against pancreatic α-amylase, α-glucosidase and lipase was examined. The antiproliferative efficacy of the plant extract against several colorectal cancer cell lines were then measured. The main flavonoids component found in Pistacia species are quercetin-3-β-D-glucoside, rutin, kaempferol and vitexin. The starch blockade IC50 (µg/mL) of Pistacia species in a descending order were: P. lentiscus leaves: 1.09±0.01; P. atlantica leaves: 0.96±0.09 and P. atlantica fruits: 0.48±0.02. Pistacia species exerted promising inhibition effect for pancreatic lipase (PL). Besides the aglycones of P. atlantica leaves, all the tested aqueous extracts exerted appreciably novel antiproliferative activity against the tested colorectal cancer cell lines. This study provides useful indication for the Pistacia species as a potential novel therapeutic agent against diabesity and cancer. [ABSTRACT FROM AUTHOR]

Diabetes mellitus (DM) is the condition represented by persistent hyperglycaemia, owing to the altered glucose metabolism. The current research aims to assess the capabilities of strains of lactic acid bacteria (LAB) that were obtained from fermented Ananas comosus (A. comosus) and Carica papaya (C. papaya) (non-dairy sources) as probiotics for the prevention of DM. The capacity of the strains to endure or survive extreme conditions was evaluated using bile tolerance using oxgall at varying concentrations, hydrophobicity, coaggregation ability, adhesion ability, and hemolytic activity, and their survival rate in gastric and intestinal juices. The LAB strains demonstrated the following results: phenol (>7 Log CFU/mL), acid-bile conditions (6.53–7.11 Log CFU/mL), gastric and intestinal juice tolerance (6.18–7.33 Log CFU/mL), thereby fulfilling the requirements to be a prospective probiotic agent. The LAB strains showed characteristics of cell surface hydrophobicity (≥ 40 %), autoaggregation (≥ 70 %), and coaggregation (≤ 48 %). Furthermore, they demonstrated antioxidant activity (≥ 59 %) as well as the ability to lower blood sugar levels by inhibiting α-glucosidase (≤ 57 %) and α-amylase (≤ 64 %). The development of antidiabetic probiotics from fermented papaya and pineapple has several potential uses. These probiotics may be used as functional foods or supplements to improve glycemic control in individuals with DM. They may also be used as adjunct therapies to conventional diabetes treatments to enhance their efficacy. The results of this research may pave way for creating probiotic products that function as food additives or supplements, with the aim of enhancing health and preventing long-term illnesses like diabetes. Additionally, probiotics have been proven to boost intestinal health, regulate the immune system, and decrease inflammation. [Display omitted] • LAB from fermented pineapple and papaya show antidiabetic potential. • LAB exhibit resilience to gastrointestinal conditions, meeting probiotic norms. • Strains display notable probiotic traits like hydrophobicity and adhesion. • Antioxidant activity and α-glucosidase & α-amylase inhibition suggest blood sugar reduction. • Pineapple and papaya offer promising non-dairy sources for functional foods. [ABSTRACT FROM AUTHOR]

Summary: To address the adverse effects of current hypoglycaemic drugs and the limited effect of individual extracts, this research investigated the inhibitory effect of combined green tea extract (GTE) and coffee (CE) on α‐amylase (α‐AMY) and α‐glucosidase (α‐GLU) and their inhibitory mechanisms. The inhibitory effects of single extracts and the complexes on α‐AMY and α‐GLU activities were compared by the principle of equivalence line. The results showed that the complexes had a good synergistic inhibition effect on both α‐AMY and α‐GLU at GTE: CE = 4:1 (w/w). Fluorescence spectroscopy and circular dichroism spectroscopy indicated that the complexes of GTE: CE = 4:1 (w/w) altered the secondary and tertiary structures of α‐AMY and α‐GLU more than the individual extracts, resulting in a synergistic inhibitory effect. The orthogonal experiments showed that tea polyphenols (TP) and coffee polyphenols (CP) were the main components responsible for the synergistic inhibitory effect of the two extracts. [ABSTRACT FROM AUTHOR]

A more efficient and environmentally friendly approach has been developed for synthesizing phosphonates through the Michaelis-Arbuzov reaction, catalyzed by nano-ZnO in a solvent-free environment, utilizing microwave irradiation. Before synthesis, each compound underwent in silico ADMET analysis and molecular docking to assess drug-like characteristics and their potential to inhibit α-amylase. The structure of the newly synthesized compounds was validated using spectroscopic analysis, and their in vitro inhibitory effects on α-amylase were assessed. Among the compounds, dimethyl 2-(anthracen-10-yl)-4-oxo-4H-chromen-6-yl-6-phosphonate (6j), dimethyl 2-(naphthalen-1-yl)-4-oxo-4H-chromen-6-yl-6-phosphonate (6h), and dimethyl 2-(1-methyl-1H-indol-3-yl)-4-oxo-4H-chromen-6-yl-6-phosphonate (6e) displayed the highest inhibitory activity compared to the reference substance, acarbose. Additionally, compounds dimethyl 2-(benzo[d][1,3]dioxol-4-yl)-4-oxo-4H-chromen-6-yl-6-phosphonate (6i), dimethyl 4-oxo-2-phenyl-4H-chromen-6-yl-6-phosphonate (6a), and dimethyl 2-(1H-indol-5-yl)-4-oxo-4H-chromen-6-yl-6-phosphonate (6g) exhibited nearly equivalent effective inhibitory activity when compared to the standard. The remaining compounds demonstrated moderate to good enzyme inhibition. [ABSTRACT FROM AUTHOR]

Background and Objectives: Malt barley is a specialty crop in which quality characteristics are as important as grain yields. Short stature genes used to increase grain yields in barley have variable effects on malt quality. Effects of sdw1.d semidwarf gene on malting quality were examined using a near isogenic line (NIL) in six field trials. Findings: sdw1.d increased malt extract percentage significantly in five trials by 1.7% and decreased test weight (2.0–3.5 kg) in three trials. Grain weight increased in three trials where lodging was common while decreasing in two where lodging was not severe and not changing in one. Diastatic power of sdw1.d NIL was significantly higher in four trials and similar in two trials compared to recurrent parent Tokak. No change was observed for α‐amylase activity. Conclusions: The lack of reports indicating a direct association of Sdw1 mutants with diastatic power and the presence of quantitative trait locus (QTL) around the Sdw1 gene in numerous studies implied that higher diastatic power of the NIL could be due to linkage rather than the pleiotropic effect of sdw1.d. Significance and Novelty: Improved malting quality of the NIL appeared to be the result of indirect effects of sdw1.d due to reduced lodging and linkage with diastatic power QTL. [ABSTRACT FROM AUTHOR]

Objective/Background: Acronychia pedunculata is commonly found in mountainous regions across Vietnam. The species has been widely used in traditional medicine to treat various ailments. Methods: The essential oils (EOs) of A. pedunculata roots, leaves, and flowers were extracted by hydrodistillation process and their chemical compositions were determined by gas chromatography-mass spectrometry (GC/MS). Besides, the EOs were assayed for in vitro α-amylase and tyrosinase inhibitory activities. In silico molecular docking simulations on the major components of the EOs were conducted. Results: The EOs from A. pedunculata roots and leaves consisted of 41 compounds (97.6 and 95.1%, respectively) while the flower EO comprised 37 compounds (98.7%). α-Pinene was found to be the most abundant component in the root and flower EOs, with the percentages of 63.9% and 52.6%, respectively. (E)-β-Caryophyllene was detected at the highest level (22.8%) in the leaf EO. The root EO had the most potent anti-α-amylase and antityrosinase activities, with IC50 values of 53.73 ± 1.94 and 815.53 ± 3.24 μg/mL, respectively. Its inhibitory activity against α-amylase was stronger than that of acarbose (88.80 ± 1.87 μg/mL). In molecular docking results, limonene was the potential compound in the EOs with the best binding affinity against tyrosinase. Moreover, the evaluation of potential interactions between the main components of the EOs and the selected proteins was elucidated. Conclusions: The findings unveiled that the EOs of A. pedunculata possessed α-amylase and tyrosinase inhibitory effects and potential applications in nutraceutical and cosmeceutical industries. [ABSTRACT FROM AUTHOR]

The digestive properties of starch are crucial in determining postprandial glycaemic excursions. Genistein, an active phytoestrogen, has the potential to influence starch digestion rates. We investigated the way genistein affected the digestive properties of starch in vitro. We performed enzyme kinetics, fluorescence spectroscopy, molecular docking, and molecular dynamics (MD) simulations for analysing the inhibitory properties of genistein on starch digestive enzymes as well as clarifying relevant mechanism of action. Our findings demonstrated that, following the addition of 10% genistein, the contents of slowly digestible and resistant starches increased by 30.34% and 7.18%, respectively. Genistein inhibited α-amylase and α-glucosidase, with half maximal inhibitory concentrations of 0.69 ± 0.06 and 0.11 ± 0.04 mg/mL, respectively. Genistein exhibits a reversible and non-competitive inhibiting effect on α-amylase, while its inhibition on α-glucosidase is a reversible mixed manner type. Fluorescence spectroscopy indicated that the presence of genistein caused declining fluorescence intensity of the two digestive enzymes. Molecular docking and MD simulations showed that genistein binds spontaneously to α-amylase via hydrogen bonds, hydrophobic interactions, and π-stacking, whereas it binds with α-glucosidase via hydrogen bonds and hydrophobic interactions. These findings suggest the potential for developing genistein as a pharmacologic agent for regulating glycaemic excursions. [ABSTRACT FROM AUTHOR]

This study investigated the activity and thermostability of α-amylase in green malt across a diverse panel of 54 barley genotypes, comprising 20 mutants, 19 hulled, 4 hulless, and 11 wild types, using starch as a substrate. The primary objective was to assess the variability in α-amylase activity among these genotypes and identify those with superior enzymatic activity and thermostability. Given that α-amylase is the most thermostable enzyme among the diastatic power enzymes yet exhibits significant activity reduction above 72.5 °C, a threshold frequently exceeded in industrial kilning and mashing. This research is therefore crucial for identifying genotypes that could enhance starch hydrolysis efficiency during mashing, a process critically dependent on sufficient enzymatic activity. We reported α-amylase activity and thermostability across a temperature range of 37 to 85 °C. The findings indicated that optimal temperature for α-amylase activity in barley malting lies between 65 and 75 °C. Interestingly, wild barley genotypes demonstrated the highest mean α-amylase activity, while hulless varieties exhibited the lowest. These results were validated by a significant negative correlation between α-amylase activity and the content of starch. Among the 54 genotypes, 11 displayed high α-amylase activity at 65 °C. Furthermore, one mutant (BL2105) and one wild genotype (WS230) exhibited high activity and thermostability at 75 °C, and another wild genotype (WS236) retained 30% of its original activity after heat treatment at 85 °C. These genotypes with enhanced α-amylase activity and thermostability could be strategically exploited in breeding programs to develop superior malt varieties. Such advancements could significantly enhance both malt quality and efficiency in beer production industry. [ABSTRACT FROM AUTHOR]

In this study, we investigated the polyphenolic composition of the n-butanol fraction of Linum trigynum L. (BELTr), a medicinal plant from the Linaceae family that grows in Algeria, using RP–UHPLC–ESI–QTOF–MS technique and evaluated of its antimicrobial, larvicidal and inhibition of α-amylase and butyrylcholinesterase (BChE) potentials. Fifty-six polyphenols were identified in the BELTr with isomers of vicenin-2 and orientin, and isovitexin as the major compounds. The extract showed a significant inhibition of BChE (IC50: 112.45 ± 3.93 µg/mL) and a good inhibition of α-amylase (IC50: 2.25 ± 4.05 mg/mL). In addition, the BFLTr exhibited antimicrobial activity against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 6538P, Salmonella enterica ATCC 13076, Bacillus subtilis ATCC 6633, Klebsiella pneumoniae ATCC 13883, Enterococcus faecalis ATCC 19433 and Pseudomonas aeruginosa ATCC 27853, as well as a yeast strain, Candida albicans ATCC 1031, with MICs values ranging between 250 and 500 µg/mL and a weak larvicidal effect. The extract's activities may be attributed to its richness in flavonoids and phenolic acids, either through a direct mechanism or in a synergistic one. [ABSTRACT FROM AUTHOR]

Abstract Diabetes is a metabolic condition defined by abnormal blood sugar levels. Targeting starch-hydrolyzing enzymes and Dipeptidyl Peptidase 4 (DPP-4) expressed on the surface of numerous cells is one of the key strategies to lower the risk of Type-2 diabetes mellitus (T2DM). Dalbergia sissoo Roxb. bark (DSB) extracts have been reported to have anti-diabetic properties. This study intended to scientifically validate use of alcoholic and hydro-alcoholic extracts of DSB for T2DM by conducting preliminary phytochemical investigations, characterising potential phytochemicals using Fourier transform infrared (FT-IR) spectroscopy and Gas chromatography-mass spectrometry (GC–MS) analysis followed by comprehensive in-silico analysis. A qualitative phytochemical evaluation indicated the presence of alkaloids, phenolics, glycosides, conjugated acids and flavonoids. Ethanolic extracts showed highest total phenolic content (TPC) (127.072 ± 14.08031 μg GAE/g dry extract) and total flavonoid content (106.911 ± 5.84516 μg QE /g dry extract). Further FT-IR spectroscopy also revealed typical band values associated with phenol, alcohol, alkene, alkane and conjugated acid functional groups. The GC–MS analysis identified 139 compounds, 18 of which had anti-diabetic potential. In-silico ADMET analysis of potential compounds revealed 15 compounds that followed Lipinski’s rule and demonstrated drug-like properties, as well as good oral bioavailability. Molecular docking was utilised to analyse their potential to interact with three targets: α-amylase, α-glucosidase, and DPP-4, which are crucial in managing diabetes-related problems. Molecular Docking analysis and membrane permeability test utilising the PerMM platform revealed that compounds in the extracts, such as Soyasapogenol B and Corydine, had better interactions and permeability across the plasma membrane than standard drugs in use. Molecular dynamics simulations also showed that selected compounds remained stable upon interaction with α-amylase. Overall, using the in-silico approaches it was predicted that DSB extracts contain potential phytochemicals with diverse anti-diabetic properties. It further needs to be investigated for possible development as formulation or drug of choice for treating T2DM.

Starch is the main source of energy for the human body. Natural products such as procyanidin (PC) and protocatechuic acid (PCA) can reduce postp'ran'dial blood sugar by controlling the digestion of starch. The inhibition effects of PC and PCA on α-amylase and α-glucosidase were studied using inhibition rate determination, Lineweaver-Burk double reciprocal diagram, ultraviolet spectroscopy, fluorescence quenching, equivalent mapping, and Fourier transform infrared spectroscopy. The results showed that the IC50 values of PC for α-amylase and α-glucosidase inhibition were 0.0524 and 0.0106 mg/mL, respectively, and those of PCA were 1.9426 and 1.0667 mg/mL respectively, which showed that PCA had better inhibition ability. The Lineweaver-Burk double reciprocal diagram showed that the inhibition of PC and PCA on α-amylase was a mixed inhibition, and the inhibition on α-glucosidase was non-competitive inhibition and competitive inhibition, respectively. The Binding of PC and PCA with α-amylase and α-glucosidase mainly depended on hydrophobic and hydrogen bonds, and further altered the enzyme structure by changing the microenvironment of amino acid residues. The inhibitory effects on α-glucosidase were the best when the ratio of PC and PCA was 1:21, and the inhibitory effects on α-amylase were optimal at 1:37. When the ratio of PC and PCA was 1:21, CI was less than 1, which showed a synergistic inhibitory effect on α-glucosidase. When the compounding ratio was 1:37, CI was less than 1, which showed a synergistic inhibitory effect on α-amylase. This study revealed the inhibitory mechanism of the combined use of PC and PCA on starch-digesting enzymes and would provide a theoretical basis for the development of natural hypoglycemic dietary supplements as health food excipients or drugs.

The purpose of this study is to explore the soil of the Gujarat, Kachchh region to identify amylase-producing bacteria and characterize them using molecular methods. The unique ecological characteristics of the Kachchh region may facilitate the isolation of these bacteria. Samples were collected from multiple locations within the Kachchh District, including Gandhidham, Rapar, Bhuj, Nakhatrana, Mandvi, and Mundra Talukas. These samples were then screened to isolate amylase-producing bacteria. A total of 27 different types of colonies were identified, out of which 16 exhibited amylase production (M1-M16). Out of 27 colonies identified, 16 showed amylase production. Strains M2, M7, and M13 exhibited high amylase activity, with M2 showing a consistent increase over 72 hours, making it a strong candidate for amylase production. Further identification of M2 stain identified M2 as a Gram-positive, spore-forming, capsulated, and motile bacillus, specifically Bacillus licheniformis. This was confirmed through DNA sequencing and analysis in the NCBI database, which showed a 99.15% similarity with Bacillus licheniformis. The study concludes that Soil in Kachchh is rich with microorganisms that produce amylase, an enzyme with diverse industrial applications. These organisms are valuable for sectors like food, textiles, paper, detergents, pharmaceuticals, and biofuel production.

Vetiveria zizanioides, known as vetiver grass, is renowned for its ecological significance in soil erosion control and the restoration of lands polluted by heavy metals. Additionally, this plant is extensively harvested for vetiver oil, widely utilized in both medicine and perfumery. The study was designed to investigate the phytochemicals, antilipase, hemoglobin antiglycation, antihyperglycemic, antifungal, and antibacterial properties of V. zizanioides. The GC-MS analysis revealed the presence of various volatile compounds, among which the most predominant ones, which are, 4,6,6-Trimethyl-2-(3-methylbuta-1,3-dienyl) −3-oxatricyclo [5.1.0.0(2,4)] octane, Cycloisolongifolene (8,9-dehydro-), and 9 H-Cycloisolongifolene (8-oxo-). It is noteworthy that this essential oil has shown significant inhibitory activity against α-amylase, α-glucosidase, pancreatic lipase, and glycation, with respective IC50 values of 0.47 ± 0.01, 0.42 ± 0.004, 0.58 ± 0.008, and 0.49 ± 0.005 mg/mL. Additionally, the antimicrobial activity of EoVZ was assessed against a diverse spectrum of Gram+ and Gram- bacteria, as well as fungi, using both disc-diffusion and broth-microdilution techniques. The results showed significant antimicrobial effects against all tested microorganisms, with inhibition zones ranging from 28.33 ± 1.66 mm to 9.0 ± 0.0 mm for bacteria and from 22.5 ± 1.7 mm to 14.0 ± 0.3 mm for fungi. Moreover, minimum inhibitory concentrations (MIC), minimum bactericidal concentrations (MBC), and minimum fungicidal concentrations (MFC) were determined, with values ranging from 0.0625% to 2.0% (v/v) for bacteria and from 2.0% to 8.0% (v/v) for fungi. The MBC/MIC and MFC/MIC ratios indicated bactericidal and fungicidal mechanisms. This study underscores the prominent attributes of EoVZ, particularly its antimicrobial and antidiabetic properties, suggesting its potential as a natural and effective agent ensuring high safety standards for public health.

The thermostable α-amylase derived from Bacillus licheniformis (BLA) has multiple advantages, including enhancing the mass transfer rate and by reducing microbial contamination in starch hydrolysis. Nonetheless, the application of BLA is constrained by the accessibility and stability of enzymes capable of achieving high conversion rates at elevated temperatures. Moreover, the thermotolerance of BLA requires further enhancement. Here, we developed a computational strategy for constructing small and smart mutant libraries to identify variants with enhanced thermostability. Initially, molecular dynamics (MD) simulations were employed to identify the regions with high flexibility. Subsequently, FoldX, a computational design predictor, was used to design mutants by rigidifying highly flexible residues, whereas the simultaneous decrease in folding free energy assisted in improving thermostability. Through the utilization of MD and FoldX, residues K251, T277, N278, K319, and E336, situated at a distance of 5 Å from the catalytic triad, were chosen for mutation. Seventeen mutants were identified and characterized by evaluating enzymatic characteristics and kinetic parameters. The catalytic efficiency of the E271L/N278K mutant reached 184.1 g L−1 s−1, which is 1.88-fold larger than the corresponding value determined for the WT. Furthermore, the most thermostable mutant, E336S, exhibited a 1.43-fold improvement in half-life at 95 ℃, compared with that of the WT. This study, by combining computational simulation with experimental verification, establishes that potential sites can be computationally predicted to increase the activity and stability of BLA and thus provide a possible strategy by which to guide protein design.

Amylases are highly needful product in food, beverage, and detergent industry. With the objective of isolating α-amylases producing strains, samples were collected from local bakery chimneys and kitchens, Chennai, Tamil Nadu, India. The potential α-amylase producers were isolated and screened by Starch-Iodine plate assay method. The optimized parameters such as 1.5% of starch concentration in the media with 7.2-pH at 35 °C for 36 h enhanced the production of α-amylase. The strain showing high potential, identified by 16s rRNA sequencing was found to be Bacillus subtilis. The Bacillus subtilis strain produced extracellular α-amylase with activity of 1330.14 U/mL. Further partial purification by ammonium sulphate precipitation followed by dialysis and column chromatography enhanced the specific activity to 1695.306 U/mg. The molecular weight of enzymes was found to be around 36 kDa. The enzyme stability and activity characterization revealed that the enzyme was thermotolerant which was able to retain more than 70% activity at 70 °C, 80 °C and 90 °C. The purified enzyme was also found to be Ca2+ independent. The hydrolysis of raw starch from rice, wheat and corn flour using the purified α-amylase from Bacillus subtilis showed significant activity of 72.8%, 76.1% and 65.2% with 4 h incubation respectively. The potentiality of the enzyme as an additive with commercial detergent was also determined to be effective. This study favourably attracts the attention towards industrial applications. [ABSTRACT FROM AUTHOR]

Abstract Background Peganum harmala L. is used in traditional medicine to treat several health ailments. Hence, the present work aimed to investigate the DPPH free radical scavenging, α-amylase, cytotoxic, and antifibrotic effects of the hydrophilic extract and fixed oil obtained from P. harmala seeds. Methods The hydrophilic extract and fixed oil of P. harmala were assessed for their abilities to scavenge DPPH free radicals and inhibit α-amylase using reference bioassays. The cytotoxicity was assessed on several cancer and normal cell lines, including B16F1, Caco-2, COLO205, HeLa, Hep 3B and Hep G2, MCF-7, and HEK-293 T cells. The MTS assay was used to evaluate the antifibrotic capabilities utilizing the human hepatic stellate (LX-2) cell line. Results P. harmala plant fixed oil has potent DPPH free radical scavenging activity with an IC50 dose of 79.43 ± 0.08 µg/ml. Besides, the hydrophilic extract has a poor anti-α-amylase effect compared with the antidiabetic drug Acarbose, with IC50 doses of 398 ± 0.59 and 25.11 ± 1.22 µg/ml, respectively. In addition, the growth of MCF-7, Hep3B, HepG2, HeLa, COLO205, CaCo2, B16F1, and HeK293t was inhibited by P. harmala hydrophilic extract with IC50 doses of 121.34 ± 1.71, 268.3 ± 0.75, 297.20 ± 1.00, 155.60 ± 1.14, 150.01 ± 0.51, 308.35 ± 0.53, 597.93 ± 1.36, and 5.38 ± 0.99 µg/ml, respectively. In addition, at 1000 µg/ml, 5-Fluorouracil reduced fibrosis cells by 0.089%, while the hydrophilic extract decreased the number of LX-2 cells by 5.81%. Conclusion P. harmala plant-fixed oil exhibits potential antioxidant properties. While the hydrophilic extract showed limited effectiveness as an anti-α-amylase agent and demonstrated notable cytotoxic effects against various tested cancer cell lines. Furthermore, this extract significantly reduces the number of LX-2 fibrotic cells. These findings emphasize the therapeutic potential of these products in managing various health disorders and warrant further investigation into their mechanisms of action and clinical applications.

Abstract Underutilised legumes are important crop species whose cultivation and consumption are declining despite their medicinal and nutritional benefits. One of the most pressing global health concerns of the twenty-first century is diabetes, characterised by elevated blood sugar levels (hyperglycaemia). Food plays a huge role in the incidence and management of metabolic diseases such as diabetes. Hence, pharmacological research along the food-medicine continuum is being ignited by recent evidence linking diet to health and illness incidence. Herein, we determined in vitro the inhibitory effects of the gene bank-stored seed and freshly cultivated seed and leaf extracts of ten accessions of an underutilised legume (Macrotyloma geocarpum) on the enzymes involved in carbohydrate breakdown to glucose (α-amylase and α-glucosidase) as a strategy to reduce hyperglycaemia. The extracts of seed accessions obtained from the gene bank showed weakly α-amylase inhibition (IC50 range = 300–1000 µg/mL) while, extracts of the cultivated seed accessions generally showed enhanced inhibitory effects (IC50 range = 63–355 µg/mL), when compared to standard Acarbose® (IC50 = 64.23 ± 0.02 µg/mL). Freshly harvested seed accessions (72–616 µg/mL) were also observed to have enhanced inhibitory effects on α-glucosidase enzyme than the seeds from the gene bank (39–185 µg/mL) when compared to Acarbose® (73.23 ± 0.02 µg/mL). However, the leaf extracts of the cultivated accessions displayed significantly higher α–amylase inhibition (IC50 range = 54–61 µg/mL) than the cultivated seeds while a reversed case was observed in the α–glucosidase inhibition. Overall, this study suggests the medicinal importance of the underutilised M. geocarpum legume in the management of diabetes while considering the varying bioactive effects of the accessions as well as cultivation and storage.

Tea tree essential oil (TEO) Rosemary Essential Oil (REO) and Guggul essential oil (GEO) (EOs) are a priceless essential oil that has been linked to several biological activities, including antibacterial, antifungal, immunomodulatory, anticancer, and anti-inflammatory effects. α-amylase inhibition is a hopeful curative target against type-2 diabetes as it can downgrade fierce digestion and absorption of carbohydrates into absorbable monosaccharides. The purpose of the study is in silico molecular docking of principal component of TEO, REO and GEO followed by and in vitro validation of inhibition of α-amylase activity. For docking Cb-dock2 tool was utilized. Ligand-Protein 2-D interactions were also studied. From the perspective of human health, in-silico ADMET pharmacoinformatic features (Physicochemical, Lipophilicity, Medicinal Chemistry, Druglikeness, Absorption, Water Solubility, Distribution, Metabolism, Pharmacokinetics, Excretion) have prospected. Using α-amylase, wet lab validation was carried out. 2, 2-Diphenyl-1-picryl hydrazyl (DPPH) radical inhibition assay was conducted to ascertain antioxidant role of all EO’s. Docking investigation demonstrated the effective binding of all the ligands with the α-amylase. The interaction results imply that the enzyme-ligand complexes form hydrogen, hydrophobic, and other interactions. In-silico ADMET examination disclosed that all the ligand molecules have no toxic effect and acceptable absorption as well. Further, TEO, REO and GEO has dose-dependent inhibitory action against α-amylase. All EO’s depicted good antioxidant potential. Kinetic analysis revealed that TEO, REO and GEO competitively inhibited α-amylase. It was concluded that these substances can function as model molecules for the synthesis of novel anti-diabetic substances.

Plants represent a great source of medicines, and for their components to be discovered, extraction processes must be developed, especially methods based on green technology. Supercritical fluid extraction (SFE) was employed as a green method for Moringa oleifera extraction in the present investigation. The maximum yield of extraction was obtained at 25 MPa. Moreover, the extraction at 25 MPa induced the release of various phenols and flavonoids, as analyzed via high-performance liquid chromatography. The investigation revealed the concentrations of chlorogenic, gallic, rosmarinic, and coumaric acids to be 150.59, 89.90, 44.75, and 29.41 µg/mL, respectively at 25 MPa. However, their concentrations were 0.73, 1.53, 0.24, and 0.04 µg/mL, respectively at 15 MPa; vs. 4.73, 2.62, 1.06, and 0.50 at 35 MPa, respectively. Totals of saponin, flavonoid, phenolic, tannins, and alkaloid were recorded in maximum yield at 25 MPa. Moringa oleifera extracted at 35 MPa reflected highest inhibition zones of 27 ± 0.1, 30 ± 0.2, and 30 ± 0.1 mm against C. glabrata, C. tropicalis, and C. albicans, correspondingly. α-Amylase and α-glucosidase activities were greatly suppressed by the M. oleifera extract at 25 MPa with less IC50 (12.97 µg/mL and 6.0 µg/mL), than the IC50 (53.46 and 22.02 µg/mL) at 15 MPa, compared with acarbose IC50 (5.52 and 2.64 µg/mL), correspondingly.

Abstract α-Amylase plays a crucial role in the industrial degradation of starch. The genus Jeotgalibacillus of the underexplored marine bacteria family Caryophanaceae has not been investigated in terms of α-amylase production. Herein, we report the comprehensive analysis of an α-amylase (AmyJM) from Jeotgalibacillus malaysiensis D5T (= DSM28777T = KCTC33550T). Protein phylogenetic analysis indicated that AmyJM belongs to glycoside hydrolase family 13 subfamily 5 (GH13_5) and exhibits low sequence identity with known α-amylases, with its closest counterpart being the GH13_5 α-amylase from Bacillus sp. KSM-K38 (51.05% identity). Purified AmyJM (molecular mass of 70 kDa) is stable at a pH range of 5.5–9.0 and optimally active at pH 7.5. The optimum temperature for AmyJM is 40 °C, where the enzyme is reasonably stable at this temperature. Similar to other α-amylases, the presence of CaCl2 enhanced both the activity and stability of AmyJM. AmyJM exhibited activity toward raw and gelatinized forms of starches and related α-glucans, generating a mixture of reducing sugars, such as glucose, maltose, maltotriose, maltotetraose, and maltopentaose. In raw starch hydrolysis, AmyJM exhibited its highest efficiency (51.10% degradation) in hydrolyzing raw wheat starch after 3-h incubation at 40 °C. Under the same conditions, AmyJM also hydrolyzed tapioca, sago, potato, rice, and corn raw starches, yielding 16.01–30.05%. These findings highlight the potential of AmyJM as a biocatalyst for the saccharification of raw starches, particularly those derived from wheat.

Starch was the main source of carbohydrates in people's daily diet, and its digestibility was an important factor affecting the body's postprandial blood glucose level. In this paper, the effect of xylose on the in vitro digestive properties of corn starch and its mechanism of action were investigated. The in vitro digestibility of starch was evaluated by an in vitro simulated digestion system after different additions of xylose (5%, 10%, and 15%), and the mechanisms of xylose on α-amylase and α-glucosidase, as well as its effect on blood glucose fluctuation through enzyme inhibition kinetics, molecular docking, and human glycemic index (GI) tests. The results showed that the digestive hydrolysis of corn starch was inhibited by different amounts of xylose, and the IC50 values of xylose on α-glucosidase and α-amylase were 205.35 and 4.75 mg/mL, respectively, which belonged to the mixed type inhibitors. Xylose interacted with amino acid residues of α-glucosidase and α-amylase through hydrogen bonding with binding energies of −3.05 and −5.30 kcal/mol, respectively. Additionally, the GI value of plant-based peptidoglycan milk decreased from 58 to 45 after the addition of xylose, which demonstrated that xylose was effective in decreasing the fluctuation of postprandial blood glucose. The results could provide scientific data to support the research and application of xylose in nutritional and health foods.

Abstract In the field of biotechnology, the utilization of agro-industrial waste for generating high-value products, such as microbial biomass and enzymes, holds significant importance. This study aimed to produce recombinant α-amylase from Anoxybacillus karvacharensis strain K1, utilizing whey as an useful growth medium. The purified hexahistidine-tagged α-amylase exhibited remarkable homogeneity, boasting a specific activity of 1069.2 U mg−1. The enzyme displayed its peak activity at 55 °C and pH 6.5, retaining approximately 70% of its activity even after 3 h of incubation at 55 °C. Its molecular weight, as determined via SDS-PAGE, was approximately 69 kDa. The α-amylase demonstrated high activity against wheat starch (1648.8 ± 16.8 U mg−1) while exhibiting comparatively lower activity towards cyclodextrins and amylose (≤ 200.2 ± 16.2 U mg−1). It exhibited exceptional tolerance to salt, withstanding concentrations of up to 2.5 M. Interestingly, metal ions and detergents such as sodium dodecyl sulfate (SDS), Triton 100, Triton 40, and Tween 80, 5,5ʹ-dithio-bis-[2-nitrobenzoic acid (DNTB), β-mercaptoethanol (ME), and dithiothreitol (DTT) had no significant inhibitory effect on the enzyme’s activity, and the presence of CaCl2 (2 mM) even led to a slight activation of the recombinant enzyme (1.4 times). The Michaelis constant (K m ) and maximum reaction rate (V max ), were determined using soluble starch as a substrate, yielding values of 1.2 ± 0.19 mg mL−1 and 1580.3 ± 183.7 μmol mg−1 protein min−1, respectively. Notably, the most favorable conditions for biomass and recombinant α-amylase production were achieved through the treatment of acid whey with β-glucosidase for 24 h.

This study aimed to evaluate the phenolic profile, antioxidant, antibacterial, and antidiabetic properties of ethanolic and methanolic extracts of the different parts of the endemic Odontarrhena floribunda. The antioxidant activities of O. floribunda were determined with 1,1-diphenyl-1-picrylhydrazyl (DPPH) and the copper-reducing antioxidant capacity (CUPRAC) assays. The strongest antioxidant activity was recorded in methanolic root extract (IC50:1.68±0.25 μg/mL) for DPPH and ethanolic root (λ450:0.68±0.20) extract for CUPRAC. Antibacterial activities of the extracts were displayed by the Resazurin Microplate Method (REMA). All plant extracts analyzed demonstrated stronger effects on Acinetobacter baumannii (ATCC 02026) than those of the standard compound. The in vitro α-amylase and α-glucosidase inhibitory of the plant extract were assessed using the Iodine method. Except for the methanol leaf extract, all other extracts showed strong α-glucosidase inhibitory activity (0.88–2.13 mmolTE/g). These results show that this endemic O. floribunda is a potential natural antioxidant, antibacterial, and enzyme inhibitor source. [ABSTRACT FROM AUTHOR]

Objective: Diabetic individuals have a higher probability of suffering from illness and death due to small blood vessel-related problems such as retinopathy, neuropathy, nephropathy, and stroke than other complications. There are many synthetic anti-diabetic agents available, but these can be expensive and have undesirable pathological effects. The enzyme α-amylase (hydrolase), catalyzes the hydrolysis of starch to maltose and glucose via the cleavage of α-1,4-glucosidic linkages. Diabetes mellitus patients may benefit from a therapeutic strategy that involves slowing the hydrolysis of starch by inhibiting the activity of α-amylase. Thus, looking for cost-effective, natural, and safe antidiabetic agents is essential. This study aims to screen phytoconstituents and evaluate the in-vitro and in-silico α-amylase inhibitory activity of the ethanolic extract of Adhatoda vasica leaves. Methods: The extraction of Adhatoda vasica leaves was performed using ethanol via the Soxhlet extraction process. Different concentrations (100 μg/mL to 1000 μg/mL) of ethanolic extract, Acarbose, and Sitagliptin, were prepared and evaluated for α-amylase inhibitory activity using the spectrophotometric method. Molecular docking (AutodockVina 1.2.0) and toxicity profiling (SToPToX web server) studies were performed. Results: The ethanolic extract of Adhatoda vasica leaves showed the highest percentage inhibition against α-amylase (56.763 ± 0.0035) at a concentration of 1000 μg/mL. The in-silico study supported this inhibitory activity. Vasicoline (C5) and Quercetin (C9), the active constitute of Adhatoda vasica, showed the best binding energies of −8.3 and −8.0 Kcal/mol, respectively against α-amylase enzyme (PDBID: 4W93). A toxicity study revealed the safety profile of the plant extract. Conclusion: It was concluded that Adhatoda vasica leaves possess some bioactive compounds that are responsible for controlling blood glucose levels, and their identification, purification, and isolation may lead to the development of new therapeutic agents with fewer side effects than the available drugs. [ABSTRACT FROM AUTHOR]

Objectives: Oxidative stress is implicated in several metabolic cascades involved in glucose control. Hence, investigating antioxidant and antidiabetic activities is crucial for discovering an effective diabetes mellitus (DM) agent. This study was aimed at probing the therapeutic efficacy of hydro-ethanolic extract of Combretum paniculatum (HECP) and gas chromatography-flame ionization detector (GC-FID)-identified phytochemicals as novel agents for DM. Methods: We determined the total phenols, flavonoids, and antioxidant vitamins in HECP using standard methods. A GC-FID was used to decipher phytochemicals of HECP. The antioxidant and antidiabetic activities of HECP were assessed using in vitro and in silico approaches. Results: The results revealed that HECP is affluent in phenols, flavonoids, and vitamin E and demonstrated engaging antioxidant activities in 1,1-diphenyl-2-picryl-hydroxyl (DPPH; IC50 = 0.83 µg/mL), thiobarbituric acid-reactive substances TBARS; IC50 = 2.28 µg/mL), and ferric-reducing antioxidant power assay (FRAP; IC50 = 2.89 µg/mL). Compared with the reference drug, acarbose, HECP exhibited good α-amylase and α-glucosidase inhibitory capacity, having IC50 values of 14.21 and 13.23 µg/mL, respectively, against 13.06 and 11.71 µg/mL recorded for acarbose. More so, the extract's top 6 scoring phytochemicals (rutin, kaempferol, epicatechin, ephedrine, naringenin, and resveratrol) had strong interactions with amino acid residues within and around α-amylase and α-glucosidase active site domains. All the compounds but rutin had favourable drug-like characteristics, pharmacokinetics, and safety profiles when compared with acarbose. Conclusion: Altogether, our results vindicate the use of this herb in treating DM locally and reveal that it has pharmaceutically active components that could be used as novel leads in the development of DM drugs. [ABSTRACT FROM AUTHOR]

Owing to the importance of α‐amylase in industry, it is necessary to develop a more simple and practical approach to measure α‐amylase activity. Herein, a fluorescence method was established for the detection of α‐amylase activity based on hydroxyproply‐β‐cyclodextrin‐curcumin inclusion complex (HP‐β‐CDs‐Cur). Curcumin was utilized as a signal probe, while hydroxyproply‐β‐cyclodextrin (HP‐β‐CDs) was employed both as a substrate of α‐amylase and the carrier of curcumin. α‐amylase is capable to hydrolyse α‐1,4‐glycosidic bond on the HP‐β‐CDs ring, facilitating the detection of its activity through the measurement of the decrease of fluorescence intensity caused by the release of curcumin into the aqueous. The results showed that there was a good linear relationship between fluorescence intensity and α‐amylase concentration in the range of 0.065~1.365 U mL−1, and the detection limit was 0.0065 U mL−1. The successful application of this method to evaluate the activities of multiple commercial α‐amylases underscores its robustness and reliability. [ABSTRACT FROM AUTHOR]

BACKGROUND RESULTS CONCLUSION Phyllanthus emblica Linn. (PE) is rich in polyphenols, which can be categorized into free and bound phenolics (PEFP and PEBP). This study evaluated the inhibitory effect of PEFB and PEBP on α‐amylase for the first time. The mechanism of the inhibition effect of PEFP and PEBP on α‐amylase was investigated by enzyme inhibition kinetics, multispectral analysis, thermodynamics, and molecular docking.Free and bound phenolics inhibited α‐amylase activity effectively in a mixed type of inhibition. Fluorescence quenching and thermodynamic analyses showed that the binding of PEFP and PEBP to α‐amylase occurred through a static quenching process (Kq = 6.94 × 10¹² and 5.74 × 10¹² L mol−1 s−1), which was accompanied by a redshift (λem from 343 to 347 nm), leading to a change in the microenvironment. This process was found to be a spontaneous exothermic reaction (ΔG < 0). Circular dichroism (CD) analysis confirms that the secondary structure of α‐amylase was altered, in particular a decrease in α‐helixes and an increase in random coils. Molecular docking studies showed that PEFP and PEBP interacted with α‐amylase through hydrogen bonding and hydrophobic interactions.The present study provides valuable insights into the mechanism of action of PEFP and PEBP on α‐amylase, which will provide a theoretical basis for their possible use as novel natural α‐amylase inhibitors. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: Peganum harmala L. is used in traditional medicine to treat several health ailments. Hence, the present work aimed to investigate the DPPH free radical scavenging, α-amylase, cytotoxic, and antifibrotic effects of the hydrophilic extract and fixed oil obtained from P. harmala seeds. Methods: The hydrophilic extract and fixed oil of P. harmala were assessed for their abilities to scavenge DPPH free radicals and inhibit α-amylase using reference bioassays. The cytotoxicity was assessed on several cancer and normal cell lines, including B16F1, Caco-2, COLO205, HeLa, Hep 3B and Hep G2, MCF-7, and HEK-293 T cells. The MTS assay was used to evaluate the antifibrotic capabilities utilizing the human hepatic stellate (LX-2) cell line. Results: P. harmala plant fixed oil has potent DPPH free radical scavenging activity with an IC50 dose of 79.43 ± 0.08 µg/ml. Besides, the hydrophilic extract has a poor anti-α-amylase effect compared with the antidiabetic drug Acarbose, with IC50 doses of 398 ± 0.59 and 25.11 ± 1.22 µg/ml, respectively. In addition, the growth of MCF-7, Hep3B, HepG2, HeLa, COLO205, CaCo2, B16F1, and HeK293t was inhibited by P. harmala hydrophilic extract with IC50 doses of 121.34 ± 1.71, 268.3 ± 0.75, 297.20 ± 1.00, 155.60 ± 1.14, 150.01 ± 0.51, 308.35 ± 0.53, 597.93 ± 1.36, and 5.38 ± 0.99 µg/ml, respectively. In addition, at 1000 µg/ml, 5-Fluorouracil reduced fibrosis cells by 0.089%, while the hydrophilic extract decreased the number of LX-2 cells by 5.81%. Conclusion: P. harmala plant-fixed oil exhibits potential antioxidant properties. While the hydrophilic extract showed limited effectiveness as an anti-α-amylase agent and demonstrated notable cytotoxic effects against various tested cancer cell lines. Furthermore, this extract significantly reduces the number of LX-2 fibrotic cells. These findings emphasize the therapeutic potential of these products in managing various health disorders and warrant further investigation into their mechanisms of action and clinical applications. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the influence of ozone (O3) as a novel pre-treatment for carrot drying. To establish the best conditions, carrot cubes were exposed to increasing concentrations of O3 (0-40 µg/L) for 2 h before air convective drying at 70 °C. According to the mass transfer models, the results highlighted a correlation between higher effective diffusivity (4.654 x 10-11 m²/s) and increasing O3 concentrations. Furthermore, dried carrot cubes exhibited sustained levels of phenolic compounds (29.33-56.42 mg GAE/100 g), carotenoids (5.17-14.58 mg/100 g), antioxidant activity (1.27-5.01 µM Trolox/g), and inhibitory activity (IC50) against α-amylase (56.02-95.63 μg/mL) and α-glucosidase (29.72-69.08 μg/mL) as the O3 concentration increased up to 40 µg/L. Differential scanning calorimetry analysis showed different thermal profiles on carrot cubes due to O3 treatment. Furthermore, the O3 pre-treatment significantly accelerated and enhanced the rehydration of carrot cubes, resulting in faster absorption of moisture and restoration to the initial texture and volume. Overall, this study shows that O3 pre-treatment (40 µg/L) can be a rational strategy to optimize drying process, bringing an innovative and straightforward solution to preserve the techno-functionality of carrots. Future research should investigate the long-term stability of bioactive compounds and sensory characteristics of dried carrot subjected to O3 pre-treatment. [ABSTRACT FROM AUTHOR]

Current antidiabetic medications have a plethora of harmful side effects, and most of the drugs do not contain 1,3,4-oxadiazole moiety. Therefore, research into the development of novel drugs which contain 1,3,4-oxadiazole moiety with fewer side effects is necessary. Novel fluorine-incorporated 1,3,4-oxadiazole derivatives (1c–1n) were synthesized, characterized, and evaluated for α-amylase and α-glucosidase enzyme inhibitor activity. An in vivo Drosophila melanogaster model and an in vitro system were used to investigate its antidiabetic properties, further, which were characterized by 1HNMR, MASS, and FT-IR. Spectroscopic techniques and DFT are used to calculate more geometrically optimized molecule structures using the B3LYP technique. The compounds were tested against the α-glucosidase and α-amylase enzymes. Among different compounds tested, compounds 1i (IC50 = 54.83 µg/ml), 1k (IC50 = 64.95 µg/ml), 1m (IC50 = 64.78 µg/ml), and 1n (IC50 = 66.30 µg/ml) showed significant α-amylase inhibitor efficacy compared to the acarbose (IC50 = 35.17 µg/ml); compounds 1m (IC50 = 74.64 µg/ml) and 1i (IC50 = 60.35 µg/ml) exhibited significant α-glucosidase inhibitory action compared to acarbose (IC50 = 46.06 µg/ml). The docking study exhibited that compound 1i creates six hydrogen bonds and compound 1m forms three hydrogen bonding contacts at the active site of the enzyme (PDBID:4w93). The obtained results are demonstrated that compounds 1i, 1m, 1n, and 1k had greater antidiabetic activities and can be further taken into consideration for antidiabetic therapeutic interventions. [ABSTRACT FROM AUTHOR]