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Glycosaminoglycans (GAG) are bioactive polysaccharide rich in -SO 3 - and -COO- groups, also known as acidic mucopolysaccharides. In this study, the feasibility of three structurally distinct forms of chondroitin sulfate (CS-A, CS-C, and CS-D) from the GAG family was explored as a potential strategy to enhance industrial α-amylase activity. All three CSs were found to increase α-amylase activity to varying degrees, with CS-D showing the most significant increase, exceeding 78 %. Furthermore, fluorescence quenching experiments indicated that the interaction between CS and α-amylase is primarily driven by hydrophobic interactions. In silico, molecular docking revealed that the sulfate groups of all three CSs form hydrogen bonds with α-amylase, with CS-D exhibiting the lowest binding energy due to its two sulfate groups. Kinetic simulations further suggested that binding to CS increases the flexibility of key active site residues (Asp197, Glu233, and Asp300), modifies the secondary structure, and enlarges the substrate-binding pocket, thereby promoting α-amylase's hydrolytic activity. Thus, this work revealed CS as an α-amylase activator and further elucidated its interaction mechanism using in vitro and in silico studies, which may be beneficial to apply CS in pharmaceutical or food industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The aim of the study was to analyze the potential health-promoting and nutritional components of leaves and figs from several Spanish varieties of Ficus carica L. The study focused to identify (by LC-PDA-QTof/MS) and quantify (by UPLC-PDA-FL) various components including carotenoids, chlorophylls, tocols, amino acids, phenolic acids, flavonols, anthocyanins. Besides, the sugar profile, the antioxidant capacity (ORAC, FRAP and ABTS) and the in vitro hypoglycaemic potential via inhibition of α-amylase and α-glucosidase were evaluated. The leaves were found to have significant antioxidant properties. This is attributed to their high content of carotenoids (3170.77-6763.77 mg/ 100 g dm), chlorophylls (405.58-744.23 mg/ 100 g dm), tocols (59.35-115.61 mg/ 100 g dm), and polyphenols (1150.34 mg/100 g dm). Additionally, figs presented greater sources of amino acids (725.07 mg/100 g dm) and exhibited higher anti-diabetic activity than leaves. Figs of "Cuello Dama Negra" variety presented the highest content of anthocyanins (108.22 mg/100 g dm). The study suggests that incorporating these vegetal materials into another food matrix could have potential health benefits, especially in terms of antioxidant and anti-diabetic effects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The objective was to evaluate the synergistic effect of ozonization and α-amylase on modifying the microstructure and paste properties of starch, using 0.00042 g of ozone/100 g of buffer for various durations. Enzymatic susceptibility was increased, achieving maximum values of 12.73 % with an 11.42 % increase in crystallinity and an average particle size of 10.12 μm for 90 min treated japonica rice starch (JR90). The granules exhibited a polyhedral shape and, with increased intensity of combined treatments, formed clusters and lost their original geometry. Apparent viscosity, rheological, and textural parameters were reduced due to the more efficient action of α-amylase on ozonized starch, as confirmed by the low gelatinization enthalpy value (7.61 J/g). Ozone proved effective in opening starch chains, partially gelatinizing granules, homogenizing the enzymatic medium, and increasing the hydrolysis rate of α-amylase in japonica rice starch., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Most studies have primarily focused on the effects of individual sources of pectin or polyphenols on starch digestibility. However, the interactions between pectin and polyphenols in digestive fluids may influence their inhibitory capacity against starch digestion by modulating α-amylase activity (αAA), a relationship that remains poorly understood. This study aims to clarify how pectin affects starch digestion when combined with mulberry fruit powder (MFP), mulberry polyphenol extract (MPE), and rutin (the main phenolic compound in MPE). Results showed that the combination of pectin and MFP initially inhibited αAA but later enhanced it. The combination of pectin and MPE consistently showed stronger inhibition of αAA than MPE alone throughout digestion; similar results were observed for the pectin-rutin combination, though the pectin-MPE pairing exhibited greater inhibition than the latter. Pectin's enhanced inhibitory effect on starch digestion may arise from its interactions with α-amylase, starch, and polyphenols through hydrophobic interactions, hydrogen bonding, and non-covalent forces., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In this study, we investigated MalS, a periplasmic α-enzyme from Escherichia coli K12, known for its unique biochemical properties related to polysaccharide utilization. Evolutionarily, MalS has inherited the glycosyl hydrolase catalytic domain from the glycoside hydrolase family 13, with the protein sequences highly conserved across Enterobacteria, including Salmonella and Shigella. MalS exhibited optimal activity at 65 °C, significantly higher than other E. coli enzymes. Although its reaction pattern resembled that of typical α-amylases, its catalytic efficiency on polysaccharides was notably lower. Intriguingly, MalS demonstrated a strong binding affinity for various glucose polymers, including β-cyclodextrin and glycogen, which significantly enhanced its thermostability. Despite full-length MalS binding strongly to glycogen, neither its N-terminal domain, predicted by AlphaFold2 to belong to the Carbohydrate-Binding Module family 69, nor the remaining parts of the enzyme showed binding affinity toward polysaccharides. Kinetic studies revealed that MalS had a 2.5-fold lower K m and 1.4-fold higher catalytic efficiency toward glycogen compared to amylopectin, which contrasts starkly with pancreatic α-amylases. However, over prolonged reactions, glycogen hydrolysis by MalS was slower than that of amylopectin. In the early initial stage, MalS predominantly degraded glycogen to maltopentaose (G5) rather than maltohexaose (G6) as usual. Taken together, these findings suggest MalS may play a role in recognizing glycogen-type polysaccharides in the bacterial periplasm during adaptation to new environments. Given the crucial role of glycogen in the survival and infection processes of pathogenic bacteria, understanding MalS's interaction with glycogen-type polysaccharides could offer valuable insights into bacterial survival mechanisms and their ability to infect hosts. KEY POINTS: • MalS has unique structure and properties but conserved among many enterobacteria • Binding of MalS with polysaccharides significantly enhanced its thermostability • Unlike other amylases, MalS showed 2.5-fold lower K m on glycogen than amylopectin., Competing Interests: Declarations. Ethics approval: This article does not contain any studies with animal or human participants performed by any of authors. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Coreopsin is a flavonoid from Coreopsis tinctoria. The inhibition of coreopsin and synergy with acarbose against α-amylase (PPA) or α-glucosidase (SCG) were explored. As a result, coreopsin exhibited stronger inhibition on PPA/SCG than that of acarbose. Combination of coreopsin (4.11 μM) with acarbose (132.77 μM) had significant synergistic effect on PPA, while combination of coreopsin (5.76 μM) and acarbose (121.7 μM) had significant synergy on SCG. Coreopsin, acarbose and acarbose-coreopsin inhibited PPA in mixed-type mode. Acarbose competitively inhibited SCG, whereas coreopsin and acarbose-coreopsin inhibited SCG in mixed-type mode. Fluorescence analysis conformed that coreopsin could synergize with acarbose by increasing the binding ability of acarbose to PPA/SCG. Compared with acarbose or coreopsin, acarbose-coreopsin complexes resulted in more conformational changes of PPA/SCG, revealing that the complexes had stronger inhibitory ability than acarbose or coreopsin alone. The detail binding information of coreopsin, acarbose or acarbose-coreopsin to PPA /SCG was revealed by computer simulation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In this study, quince soluble dietary fiber (SDF) and insoluble dietary fiber (IDF) were obtained by acid extraction, enzyme extraction and alkaline extraction methods. The acid extracted DF displayed higher results compared to enzyme and alkaline extraction methods in terms of water holding capacity (15.97 g/g SDF), oil holding capacity (1.05 g/g SDF) and nitrite ion adsorption capacity (92.83 mg/g SDF). The antioxidant activity and phenolic content of acid extracted IDF were significantly higher than the other quince DFs. In addition, quince DFs exhibited in vitro hypoglycaemic activity, exhibiting high glucose adsorption capacity (237 mg/g) and α-amylase inhibition activity (82 %). Similarly, acid extracted SDF of quince showed in vitro hypolipidemic activity, with cholesterol adsorption capacity of 155 mg/g and lipase inhibition activity of 36 %. The structures and thermal properties of quince DFs were characterized by FT-IR and TGA. Quince DFs with high functional properties might be suitable agents for functional food formulations, such as meat products, low-calorie fruit bars, flour mixtures, etc., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Hawk tea, a conventional herbal beverage, is renowned for its beneficial properties in enhancing digestion and mitigating hyperglycemic tendencies. However, the extraction methodology for hawk tea polyphenols (HTP) has been understudied thus far, impeding its progress and broader application. To develop an efficient approach for HTP extraction, the present study introduced and optimized the application of ultrasonic-assisted aqueous two-phase extraction. Under optimal extraction conditions, the extraction yield of polyphenols from raw HTP was 7.86 %. During purification, LX-B14 was selected due to the highest adsorption and desorption abilities. Then in an in vitro simulated digestion system, HTP significantly reduced the expected glycemic index, raised the content of resistant starch, and decreased the activities of α-amylase and α-glucosidase, indicating its potential for alleviating starch digestion. Accordingly, the results provide an alternative approach for efficiently obtaining phenolic compounds from hawk tea, facilitating the advanced utilization of HTP within the food industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Postharvest processing plays a crucial role in harnessing the benefits of prickly pear fruit by utilizing betalain as natural colorants to replace artificial colors in model food systems. Prickly pear betalain-enriched gummies were developed using various sugar substitutes, including table sugar, xylitol, stevia, and fructo-oligosaccharides (FOS). These gummies were analyzed for in vitro enzymatic inhibition, anti-inflammatory effects and molecular docking studies for decoding diabetes type-II and skin resilience. FTIR and HPLC confirmed the presence of betalain and isorhamnetin across all gummies. FOS and stevia incorporated gummies exhibited the highest polyphenolics and antioxidant activity. Betalain extract combined with stevia and FOS showed significant in vitro enzyme inhibition compared to other studied gummies. Specifically, FOS gummies showed the highest inhibition rates for α-amylase (23.58 %), α-glucosidase (24.12 %), tyrosinase (54.68 %), and collagenase (2.38 %). Additionally, all samples were non-toxic to RAW cell lines and exhibited anti-inflammatory effects. Molecular docking studies corroborated the in vitro results, and pharmacokinetics profiling confirmed the gummies' suitability for oral consumption and skin safety. The developed prickly pear betalain-enriched gummies, particularly those formulated with fructo-oligosaccharides and stevia, demonstrate significant potential as functional supplements for managing diabetes type-II and skin-related conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The thermostability of enzymes plays a significant role in the starch hydrolysis process in the industry. The structural difference between thermostable Bacillus licheniformis α-amylase (BLA) and thermolabile Aspergillus niger α-amylase (ANA) is interesting to be explored. This work aimed to study the thermostability-determining factor of BLA as compared to a non-thermostable enzyme, ANA, using molecular dynamics (MD) simulation at a high temperature. A 100 ns of classical MD, which was followed by 200 ns accelerated MD was conducted to explore the conformational changes of the enzyme. It is revealed that the intramolecular interactions through salt bridge interactions and the presence of calcium ions dominates the stability effect of BLA, despite the absence of a disulfide bond in the structure. These results should be useful in designing a thermostable enzyme that can be used in industrial processes.Communicated by Ramaswamy H. Sarma.

Type 2 diabetes accounts for the largest percentage of all diabetic cases worldwide. Cucurbitane-type triterpenes are mainly found in Momordica charantia and possess excellent pharmacological activities. This study was designed to identify cucurbitane-type triterpene from Momordica charantia using Liquid Chromatography-Mass Spectrometry (LC-MS) analysis, examine its anti-diabetic property with molecular docking against diabetes enzymes (alpha-amylase, alpha-glucosidase, dipeptidyl dipeptidase IV and peroxisome proliferator-activated receptor gamma). The stability and interactions of the docked complexes were investigated using molecular dynamics simulation, while the pharmacokinetic and toxicity profile of the ligand was examined using an ADMET server. (23E)-Cucurbita-5,23,25-triene-3,7-dione (CUB) was identified from the LC-MS profiling of the methanolic extract of M. charantia . The molecular docking studies showed that the identified phytochemical elicited good binding energy against all the target receptors. The RMSD and RMSF plots obtained from the 100 ns molecular dynamics simulation showed that the ligand was stable and established substantial interactions with the amino acid residues of the diabetes enzymes which were confirmed by the MM\GBSA computations. The pharmacokinetic and toxicity properties of the ligand showed it was safer as an anti-diabetic drug candidate. Extensive isolation, in vitro and in vivo studies of the ligand against the diabetic enzymes is recommended.Communicated by Ramaswamy H. Sarma.

In this study, we adopted the synergistic modification technology of hydrodynamic cavitation and snail enzyme, to improve the yield and activity of soluble dietary fibers (SDFs) of rice husk. The physicochemical properties, structural changes, and inhibition of α-glucosidase and α-amylase of SDFs were examined in vitro. This synergistic treatment significantly increased the yield of SDFs to 18.64 % ± 0.16 %, significantly reduced the particle size to 122.33 ± 0.26 nm, and significantly increased the specific surface area to 1.718 ± 0.002 m 2 /g. The absolute value of the zeta potential significantly increased to -36.39 ± 0.12 mV, indicating an excellent solution stability and gel-forming ability. At the same time, the water-holding, oil-holding, and swelling capacities were significantly enhanced, reaching 8.52 ± 0.09 g/g, 4.85 ± 0.29 g/g, and 7.29 ± 0.25 mL/g, respectively. Structural analysis showed that the synergistic treatment destroyed the fiber structure, produced a large number of small molecule fragments, and significantly changed the monosaccharide components and functional group distribution. Functional evaluation showed that the inhibitory effect of CE-SDF on α-glucosidase and α-amylase was significantly enhanced, and enzymatic reaction kinetic analysis revealed that both enzymes were competitive inhibitors, with IC 50 values of 2.893 and 1.727 mg/mL, respectively. In summary, the synergistic modification of hydrodynamic cavitation and snail enzyme greatly optimized the structural and functional properties of rice husk SDFs, providing a theoretical basis for its application in the field of hypoglycemic drugs and functional foods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Author(s). Published by Elsevier B.V. All rights reserved.)

Characterization and comparison of the chemical compositions and biological activities of Lactarius volemus of three different sizes (large size, medium size, small size) were investigated in this work. By comparing three sizes of Lactarius volemus, the small-sized Lactarius volemus had the highest content of crude fiber, fat, crude protein, total sugar, reducing sugar, polyphenols, and total flavonoids, whereas medium-sized Lactarius volemus showed the highest content of ash and total amino acids. Meanwhile, the small-sized Lactarius volemus also exhibited the best 2,2-Diphenyl-1-picrylhy drazyl (DPPH) radical scavenging ability, α-amylase inhibitory activity, α-glucosidase inhibitory activity, and xanthine oxidase inhibitory activity. The small-sized Lactarius volemus had the highest chemical composition content and the best biological activities among the three different sizes. This work might provide basic data support for guiding the development of Lactarius volemus and also instruct consumers in selecting and purchasing high-quality Lactarius volemus., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Amylopectin (AP)-only (APBS), normal (NBS), and amylose (AM) only (AOBS) barley starches were selected here to investigate catalysis pattern of maltogenic α-amylase (MA) on hydrolyzing AP and AM granular starches. MA shortened starch side chains with degree of polymerization (DP) 11-30. MA-treated APBS exhibited porous granular structures and dramatically increased degree of branching (DB, 17-20 %), and reduced ordered degrees, suggesting high hydrolysis and transglycosylation activities of MA. MA-treated NBS showed less pronounced porous structures and slightly increased DB (2-4 %), indicating high hydrolysis but low transglycosylation activities. AOBS displayed minimal changes in DB (0.2-0.3 %) and starch structures, implying low hydrolysis and transglycosylation activities. Therefore, MA preferred to attack the AP molecules with abundant glucan substrates with DP 11-30, while AM restricted MA activity likely by creating ineffective binding sites and undergoing rapid reorganization. These findings deepened the understanding of the mechanisms of MA in modifying granular starches with varying AM content., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The continuous evolution of bacteria and the formation of biofilm have exacerbated resistance issues, highlighting the urgent need for new antibacterial materials. In this study, L-fucose was polymerized to synthesize thiolated poly(2-(L-fucose) ethyl methacrylate) (PFEMA-SH), which was subsequently co-modified with α-amylase onto gold nanorods (GNR) to prepare the antibacterial nanoparticle composite, GNR-Amy-PFEMA (G-A-P). These nanomaterials exhibit both photothermal and enzymatic properties, enabling G-A-P to effectively sterilize and disperse biofilm. Under near-infrared light irradiation, the temperature of G-A-P composite increases significantly, leading to bacterial cell damage and biofilm disruption. The G-A-P composite demonstrated nearly 100 % eradication of planktonic bacteria after 5 min of irradiation and achieved a 70.9 % reduction in mature biofilm biomass, with a 3.37-log decrease in the number of bacteria within the biofilm. These composites display strong antimicrobial activity and hold great potential for the removal of Pseudomonas aeruginosa biofilm. Furthermore, the ability of G-A-P to reduce biofilm formation without the use of traditional antibiotics suggests that it may offer an antibiotic-free alternative for managing biofilm-related infections., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cold-active pullulanases with good catalytic performance possess promising applications in cold hydrolysis of starch. Adopting bioinformatics-assisted mining strategies, 7 candidate cold-active pullulanases were initially screened out from IMG/MER database. Among the candidates, PulBs exhibited good thermostability and the highest specific activity of 147.4 U/mg. The half-life of PulBs was about 200 h at 35 °C. Employing PulBs as the initial enzyme, the active-site design of FuncLib was implemented to enhance the activity. The design PulBs-20 exhibited an enhanced specific activity of 209.9 U/mg, which was 1.4 times that of PulBs. Furthermore, the thermostability of PulBs-20 was augmented, with a half-life of 250 h at 35 °C. When applied in the cold hydrolysis of starch, PulBs-20 can effectively enhance the hydrolysis effect of raw starch. Supplemented with the raw starch-hydrolyzing α-amylase AmyZ1 and PulBs-20, the hydrolysis rate of raw corn starch increased to 53.5 %, which was 1.3 times that of using AmyZ1 alone. Due to its high hydrolysis activity and good thermostability, PulBs-20 can serve as an efficient accessory enzyme in starch cold hydrolysis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

α-Amylases, constituting a significant share of the enzyme market, are mainly synthesized by the genus Bacillus. Enzymes tailored for specific industrial applications are needed to meet the growing demand across a range of industries, and thus finding new amylases and optimizing the ones that already exist are extremely important. This study reports the successful expression, characterization and immobilization of P. megaterium α-amylase (PmAmy) in E. coli protein expression systems. The recombinant PmAmy has a molecular weight of 56 kDa and its in silico predicted model structure presents a monomer composed of three domains, like most amylases. Regarding long-term storage, PmAmy remained 60 % active after 6 weeks of storage at -20 and -80 °C indicating its stable storage at low temperatures. PmAmy was found to be Ca 2+ ion-independent for both catalytic activity and thermostability while Mn 2+ enhanced activity in a concentration-dependent manner. The optimum characteristic working conditions of PmAmy were measured as pH 7.0 and 40 °C. Immobilizing PmAmy significantly improved its thermal stability, increasing its resistance to thermal denaturation by at least 4.1-fold. Kinetic analyses revealed that the K M and V max values of free PmAmy were 0.1 mg mL -1 and 556 U mg -1 , respectively while immobilization resulted in an increase for both the K M and V max values. Kinetic analysis revealed enhanced activity for the Ca 2+ -independent immobilized enzyme, making it suitable for industrial applications particularly starch processing requiring moderate thermostability without the need for Ca 2+ ions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Inc. All rights reserved.)

α-Amylases are the workhorse enzymes of starch degradation. They are central to human health, including as targets for anti-diabetic compounds, but are also the key enzymes in the industrial processing of starch for biofuels, corn syrups, brewing and detergents. Dissection of the activity, specificity and stability of α-amylases is crucial to understanding their biology and allowing their exploitation. Yet, functional characterization lags behind DNA sequencing and genomics; and new tools are required for rapid analysis of α-amylase function. Here, we design, synthesize and apply new branched α-amylase activity-based probes. Using both α-1,6 branched and unbranched α-1,4 maltobiose activity-based probes we were able to explore the stability and substrate specificity of both a panel of human gut microbial α-amylases and a panel of industrially relevant α-amylases. We also demonstrate how we can detect and annotate the substrate specificity of α-amylases in the complex cell lysate of both a prominent gut microbe and a diverse compost sample by in-gel fluorescence and mass spectrometry. A toolbox of starch-active activity-based probes will enable rapid functional dissection of α-amylases. We envisage activity-based probes contributing to better selection and engineering of enzymes for industrial application as well as fundamental analysis of enzymes in human health., (© 2024 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

The interfacial behavior of soy hull polysaccharide (SHP) at the oil-water interface and the stabilization mechanism of high internal phase emulsion (HIPE) with three enzymes (α-amylase, trypsin and papain) were investigated. The diameter of the α-amylase-treated emulsion was the minimum at 40 min, indicating that the carbohydrate portions of SHP form a thick layer on the surface of the droplet to prevent aggregation. Moreover, Raman spectroscopy revealed significantly higher levels of disordered content of SHP emulsion treated with α-amylase at 60 min, potentially affecting the directional movement of SHP molecules in the emulsion. Conversely, the content of β-sheet and β-turn was lower than trypsin and papain, possibly due to ion-dipole interaction between the polar group residues within SHP and ions, or protonation with H + ., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2024. Published by Elsevier Ltd.)

This study focuses on evaluating new methods for the green extraction of saponin compounds from Polygonatum cyrtonema Hua (PCH). This study utilized a combination of carboxylic acid-based natural deep eutectic solvents (NADES) and various extraction techniques including conventional heat reflux-, ultrasound-, and microwave-assisted extraction. The primary objectives were to assess total saponin yield, antioxidant capacity, and enzyme inhibition efficiency. Additionally, the solvents and extracts were evaluated for their antibacterial activity. Oil-in-water (O/W) emulsions of NADES extracts were also characterized and analyzed for stability. Results indicated that three NADES systems were effective in extracting saponins, with choline chloride and lactic acid (ChCl-LA) system being the most efficient. The ChCl:LA extract exhibited antimicrobial and antioxidant activities superior to conventional organic solvent extracts. Additionally, it demonstrated maximum inhibitory activity (IC 50 values: 0.98 ± 0.03 and 1.46 ± 0.07 mg/mL, respectively) against α-glucosidase and α-amylase. The NADES extract as an aqueous phase significantly improved the stationarity of the O/W emulsion. Collectively, the study highlights the antimicrobial and technological advantages of NADES as a potential solvent for extracting saponin compounds from PCH., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Ficus auriculata Lour. (Moraceae) is an underutilized wild edible fruit widely consumed for its nutritional properties. The present study aimed to determine the phytochemical composition and in vitro antioxidant, enzyme inhibitory, anti-inflammatory and anti-cancerous properties of the F. auriculata fruit extracts through in vitro digestion (oral, gastric and intestinal phases). The extracts were obtained by hot extraction and cold maceration methods using aqueous and methanolic solvents. Major phytoconstituents identified through LC-MS was subjected to molecular docking against the target proteins. The elemental analysis shows the presence of major elements; high levels of total phenolics (124.61 ± 0.82 mg gallic acid equivalent/g), flavonoids (76.38 ± 0.82 mg quercetin equivalent/g), vitamin E (32.48 ± 0.09 mg alpha-tocopherol equivalent/g), and carbohydrate (34.59 ± 0.45 mg glucose equivalent/g) in hot extracted methanolic undigested extract (HEM UD) and high level of total protein (124.71 ± 0.34 mg bovine serum albumin equivalent/g) in cold extracted methanolic undigested fruit extract were found. HEM UD showed high antioxidant activity in 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid), 2,2-diphenyl-1-picryl-hydrazyl, and superoxide radical scavenging assays with IC 50 of 53.30 ± 0.57, 80.69 ± 0.12, and 65.47 ± 1.13 μg/mL, respectively. The HEM UD extract also potentially inhibited the enzyme activity of α-amylase, α-glucosidase, tyrosinase, and protein denaturation (IC 50 of 67.76 ± 1.22, 83.18 ± 1.23, 87.24 ± 1.15, and 65.76 ± 0.60 μg/mL). The most potent extract (HEM UD) was studied for its anticancer effects by MTT assay against the MCF-7 and HeLa cell lines and showed the IC 50 of 89.80 ± 0.56 and 60.76 ± 0.04 μg/mL, respectively. The LC-MS analysis elucidated ten phytoconstituents. Based on the molecular docking study, querciturone could potentially be an effective constituent in treating diabetes and inflammation-related issues. The findings indicated the ability of F. auriculata fruits as a promising functional food., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The human dietary system, which contains a variety of compounds such as polyphenols and polysaccharides, is very complex. Whether polysaccharides affect the inhibitory of polyphenol mixtures on α-amylase needs to be further investigated. The aim of this study was to analyze the effect and mechanism of pectin on the inhibition of α-amylase by a mixture of rutin and quercetin (R-Q). Results revealed that the inhibition and quenching affinity of R-Q for α-amylase was enhanced by pectin. The Stern-Volmer quenching constant of R-Q-α-amylase was increased by pectin from (6.08 ± 0.453) × 10 3  mL/mg to (9.80 ± 0.285) × 10 3  mL/mg. Pectin enhanced the ability of R-Q to inhibit α-amylase for two main reasons. On the one hand, it was owing to the binding of pectin to rutin, which increased the opportunity for quercetin to bind to the active center of α-amylase, thus enhancing the inhibitory effect of R-Q on α-amylase. On the other hand, pectin and quercetin simultaneously bound to different sites of α-amylase by noncovalent interactions to form the ternary complex of pectin-α-amylase-quercetin. The conformation of α-amylase and the hydrophobicity of amino acid residues were altered by the ternary complex, thereby enhancing the hydrogen bonding in the reaction system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The V-type porous starch (VPS) displays remarkable adsorption capability, and has potential for encapsulating guest compounds within its unique single helical cavity. In this investigation, VPS was produced via a "one-pot method" utilizing thermostable α-amylase in a hot ethanol solution. The study demonstrates that the activity of thermostable α-amylase remains high, reaching up to 86 % when employing a 40-50 % ethanol concentration, and up to 74 % with ≤70 % ethanol concentration. Furthermore, the enzyme exhibits robust stability at 90 °C for up to 10 h of reaction time. The resultant VPS exhibits enhanced V-type crystallinity and superior adsorption capacity compared to conventional A-type porous starch (APS). Notably, the hydrolysis of normal maize starch (NMS) by thermostable α-amylase in a 50 % ethanol solution at 90 °C yields 49.36 % VPS, which manifests a densely porous distributed structure. Additionally, the VPS is characterized by superior oil adsorption capacity (253.11 %), specific surface area (38.89 m 2 /g), and total pore volume (0.147 cm 3 /g)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Xing Zhou represents all authors., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

α-amylase is responsible for shortening the shelf life of food by degrading starch and glycogen into maltose, disaccharides, and oligosaccharides. This study focuses on the effect of atmospheric cold plasma on the activity, stability, enzyme kinetics, and structural change for its application in α-amylase modifications. A reduction in the residual activity from 96.01 % at 170 V to 88.01 % at 230 V after 12 min of treatment was observed in plasma-treated α-amylase compared to the untreated α-amylase at pH 5, 40 °C, after 120 min of reaction time using 2 g 100 mL -1 starch. An increased Km with a reduction in Vmax, kcat, and kcat/Km was observed with increasing voltage and treatment time. An increase in free sulfhydryl content, a decrease in tryptophan fluorescence, an increase in α-helix and β-sheet content, a decrease in β-turn and random coil structures, an increase in surface hydrophobicity, and protein aggregation indicate the structural changes of α-amylase were mainly due to the oxidative effects of reactive plasma species. No change in amide I, II, and III groups suggested plasma treatment did not alter the primary structure of α-amylase. The alteration in secondary, and tertiary structure after plasma treatment resulted in a loss of α-amylase activity., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study investigates the potential of a quercetin-based emulsion system to moderate starch digestion and manage blood glucose levels, addressing the lack of in vivo research. By enhancing quercetin bioaccessibility and targeting release in the small intestine, the emulsion system demonstrates significant inhibition of starch digestion and glucose spikes through both in vitro and in vivo experiments. The system inhibits α-amylase and α-glucosidase via competitive and mixed inhibition mechanisms, primarily involving hydrogen bonds and van der Waals forces, leading to static fluorescence quenching. Additionally, this system downregulates the protein expression and gene transcription of SGLT1 and GLUT2. These findings offer a novel approach to sustaining glucose equilibrium, providing a valuable foundation for further application of quercetin emulsion in food science., Competing Interests: Declaration of competing interest We have no conflict of interest to declare., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A large amount of antibiotics enters the soil environment and accumulates therein as individuals and mixtures, threatening the soil safety. However, there is little information regarding the influence of single and mixed antibiotics on key soil proteins at molecular level. In this study, setting sulfadiazine (SD) and tetracycline hydrochloride (TC) as the representative antibiotics, the interactions between these agents and α-amylase (an important hydrolase in soil carbon cycle) were investigated through multi-spectroscopic approaches, X-ray photoelectron spectrometry, and molecular modeling. It was found that both SD and TC spontaneously bound to α-amylase with 1:1 stoichiometry mainly via forming stable chemical bonds. The interactions altered the polarity of aromatic amino acids, protein backbone, secondary structure, hydrophobicity and activity of α-amylase. The SD-TC mixtures were designed based on the direct equipartition ray to comprehensively characterize the possible concentration distribution, and interactive effects indicated that the mixtures antagonistically impacted α-amylase. These findings reveal the binding characteristics between α-amylase and typical antibiotics, which probably influence the ecological functions of α-amylase in soil. This study clarifies the potential harm of antibiotics on soil functional enzyme, which is significant for the environmental risk assessment of antibiotics and their mixtures., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Nowadays α-amylase is widely used in various fields. Therefore, in this study, the effects of neutral (T 0 ), negatively charged (T 8 - ) and positively charged (T 9 + ) peptides on α-amylase activity were investigated by means of an applied protein electric field, and spectroscopy and molecular dynamics were employed to investigate this mechanism. It was found that the nature of the charge of the peptides had a strong influence on α-amylase activity, with T 8 - and T 9 + increasing and decreasing α-amylase activity, respectively, whereas T 0 had no effect on enzyme activity. Fluorescence spectroscopy and circular dichroism results indicated that the charged peptides changed the conformation of α-amylase. Meanwhile, the molecular dynamics results showed that the charged peptides changed the distribution of the surface charge of α-amylase mainly through electrostatic force, which not only changed the conformation of the enzyme, but also altered the microenvironment of the enzyme active centre, which caused α-amylase to become compact or loose to affect the enzyme activity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Protein hydrolysates with antioxidant potential have been reported to act as adjuvants in preventing and treating type-2 diabetes (T2D). This work investigated the biochemical, antidiabetic, antioxidant potential, and physicochemical properties of chia meal protein hydrolysate (CMPH). Bands smaller than 14 kDa were observed in the electrophoretic profile. The predominant amino acids were hydrophobic and aromatic. CMPH had the potential to inhibit α-amylase (IC 50 : 1.76 ± 0.13 mg/mL), α-glucosidase (IC 50 : 0.42 ± 0.13 mg/mL), and DPP-IV (IC 50 : 0.46 ± 0.14 mg/mL). Antioxidant activity for ABTS (IC 50 : 0.236 mg/mL), DPPH (8.83 ± 0.52%), and ORAC (IC 25 : 0.115 mg/mL). Against chia meal protein isolate (CMPI), CMPH has a broad solubility (pH 2-12.46). Particle size (624.5 ± 247.3 nm), low PDI (0.22 ± 0.06), ζ-potential (-31.1 ± 2.5 mV), and surface hydrophobicity (11,183.33 ± 2024.11) and the intrinsic fluorescence peak of CMPH was lower than that of CMPI. CMPH represents an alternative to add value to the agri-food co-product of the chia seed oil industry, generating food ingredients with outstanding antidiabetic and antioxidant potential., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The low-grade matcha is rich in insoluble dietary fiber. Trichoderma viride was used to increase the soluble dietary fiber to improve its functional properties. The soluble dietary fiber content increased from 6.74% to 15.24%. Pectin, hemicellulose, maltose, d-xylose, and glucose contents increased by 63.35% and 11.54%, 2.18, 0.11, and 7.04 mg/g, respectively. Trichoderma viride fermentation disrupted the dense structure of insoluble dietary fiber, resulting in a honeycomb structure and improving crystallinity by 22.75%. These structural changes led to an improved cation exchange capacity from 1.69 to 4.22 mmol/g, an increase in the inhibitory effect of α-amylase from 47.38% to 72.04%, and a 2.13-fold in the ferrous ion scavenging ability, and the IC 50 values of superoxide anion was reduced from 7.00 to 1.54 mg/mL, respectively. Therefore, Trichoderma viride fermentation is an excellent method for improving the quality of dietary fiber in matcha processing by-products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Understanding the mechanism underlying thermostabilization in naturally stable enzymes and enhancing the thermostability of unstable enzymes are crucial aspects in enzyme engineering. Despite the development of various engineering methods, there remains substantial scope for improvement. In this study, a novel concept termed as the "short board" theory is proposed, which conceptualizes proteins as barrels with each component representing a jagged board. Notably, optimizing modifications to the shortest board yields optimal enhancements in terms of thermostability performance. To validate this theory, α-amylase, an industrial bulk enzyme with multiple domains, is employed as a model enzyme. The existence of "short boards" and their impact on thermostability modification are demonstrated at the domain, residue, and atomic levels through experimental confirmation using domain substitution. Furthermore, a novel thermostable design and prediction model called Zero-Shot Hamiltonian (ZSH) is established and evaluated on α-amylase. This coevolutionary approach based on thermostability and deep learning exhibits remarkable success exclusively when applied to enzymes with fixed short boards. The integration of the "short board" theory with the ZSH model presents an innovative tool for enhancing enzymatic thermostability., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Diabetes mellitus (DM) a metabolic disorder characterized by high blood sugar levels causing damage to various organs over time. Current anti-diabetic drugs have limitations and side effects, prompting a search for new inhibitors targeting the α-amylase enzyme. This study aims to discover such inhibitors from thirty isoxazole derivatives of usnic acid using in silico approaches. The potential inhibitory effects of compounds were investigated using ADMET, molecular docking, molecular dynamic simulation, principal component analysis and density functional theory studies. ADMET analysis exhibited a wide range of physicochemical, pharmacokinetic, and drug-like qualities with no significant side effects which were then investigated using molecular docking experiment to determine the lead compound with the best binding affinity for the α-amylase enzyme. All compounds showed good binding affinity against α-amylase enzyme (-7.9 to -9.2 kcal/mol) where compound-13 showed the best binding affinity of -9.2 kcal/mol forming hydrogen bonds with Leu162, Tyr62, Glu233 and Asp300 amino acids. Furthermore, the binding posture and the stability of the compound-13-α-amylase enzyme complex was confirmed by molecular dynamic simulation experiment. Moreover, compound-13 showed binding energy value of -27.92 ± 5.61 kcal/mol, which indicated it could be an α-amylase inhibitor. Additionally, the reactivity of compound-13 was further confirmed by density functional theory analysis. The above findings suggest compound-13 to be a potential α-amylase inhibitor in DM. And setting the stage for further in vitro and in vivo experimental validation., Competing Interests: Compliance with ethical standards Conflict of interest The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In this study, two high-performing α-amylase-producing strains, CK3-5 and A8-1 were successfully isolated and characterized, which were taxonomically confirmed as Bacillus velezensis through whole-genome sequencing and bioinformatics. Bioinformatic sequence analysis and molecular docking revealed the catalytic triad (Asp173-Glu208-Asp274) essential for α-amylase function. Through metabolic engineering, the recombinant strain BAX-5/PT17amy(A8-1)SP 002 was developed, which exhibited the highest α-amylase activity of 1440 U/mL upon fermentation optimization, marking a 9.2-fold enhancement over the wild-type strain A8-1, and it successfully degraded 6 % of the starch in the tobacco leaves within 48 h, while the content of 13 harmful substances, including acetamide, pyridine, and acetonitrile, was reduced by 8.6 % to 25.2 %. This study reveals a novel α-amylase gene from B. velezensis and establishes an efficient expression system in B. amyloliquefaciens, offering valuable insights for industrial α-amylase production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Grain sorghum has emerged as a promising source for producing alternative proteins, yet current extraction methods lack efficiency. In this study, a novel enzymatic approach using α-amylase and cellulase on sorghum materials was developed to address this challenge. Comparisons were made among the proteins isolated from dry-milled sorghum flours, wet-milled sorghum gluten meals, and sorghum dried distiller's grains (DDG). Remarkably, proteins obtained from sorghum gluten meals demonstrated the highest protein purity (83-85 %) and recovery rate (92-93 %), followed by those from sorghum flour (purity 75-76 %) and DDG (purity 45-50 %). Physicochemical properties and functionalities of the isolated sorghum proteins were analyzed and compared with common commercial plant proteins (e.g., soy protein isolate, pea protein isolate, and wheat gluten). Sorghum proteins exhibited higher levels of crude fat content, α-helix, and random coil structures, along with higher surface hydrophobicity and oil holding capacity (OHC) compared to the commercial plant protein isolates. Notably, proteins extracted from sorghum flours displayed slightly higher α-helix and random coil structures, total sulfhydryl content, water holding capacity (WHC), OHC, and protein digestibility compared to proteins isolated from sorghum gluten meals. Overall, this study demonstrates that enzymatic processing is feasible in producing sorghum proteins and provides insights into their basic properties and functionalities., Competing Interests: Declaration of competing interest The authors declare that they have no known conflict of interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In this study, the structural characterization, physicochemical properties, antioxidant, hypolipidemic, and hypoglycemic potentials of polysaccharide components (BLP-1, BLP-2, and BLP-3) purified from blueberry leaf polysaccharides (BLP) were investigated. Ion chromatography results showed that BLP-1, BLP-2, and BLP-3 contained rhamnose, arabinose, galactose, glucose, and glucuronic acid. In contrast to BLP-1, BLP-2 and BLP-3 included galacturonic acid. The methylation analysis results indicated that the backbones of BLP-1, BLP-2, and BLP-3 were mainly composed of glycosidic linkages of arabinose, galactose, and glucose, which was consistent with the results of the previously determined monosaccharide composition. The in-vitro antioxidant results showed that BLP-1, BLP-2, and BLP-3 possessed antioxidant activity with the highest scavenging of -OH radicals. Furthermore, BLP-1, BLP-2, and BLP-3 showed high bile acid-binding activity and α-amylase inhibitory activity, suggesting that they have the potentials of hypolipidemic and hypoglycemic. This study provides a reference for the utilization of blueberry leaf resources., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Research on the co-production of multiple enzymes by Bacillus velezensis as a novel species is still a topic that needs to be studied. This study aimed to investigate the fermentation characteristics of B. velezensis D6 co-producing α-amylase and protease and to explore their enzymatic properties and applications in fermentation., Results: The maximum co-production of α-amylase and protease reached 13.13 ± 0.72 and 2106.63 ± 64.42 U mL -1 , respectively, under the optimal fermented conditions (nutrients: 20.0 g L -1 urea, 20.0 g L -1 glucose, 0.7 g L -1 MnCl 2 ; incubation conditions: initial pH 7.0, temperature 41 °C, 8% inoculation size and 30% working volume). Moreover, the genetic co-expression of α-amylase and protease increased from 0 to 24 h and then decreased after 36 h at the transcriptional level, which coincided with the growth trend of B. velezensis D6. The optimal reaction temperature of α-amylase was 55-60 °C, while that of protease was 35-40 °C. The activities of α-amylase and protease were retained by over 80% after thermal treatment (90 °C, 1 h), which indicated that two enzymes co-produced by B. velezensis D6 demonstrated excellent thermal stability. Moreover, the two enzymes were stable over a wide pH range (pH 4.0-8.0 for α-amylase; pH 4.0-9.0 for protease). Finally, the degrees of hydrolysis of corn, rice, sorghum and soybeans by α-amylase from B. velezensis D6 reached 44.95 ± 2.95%, 57.16 ± 2.75%, 52.53 ± 4.01% and 20.53 ± 2.42%, respectively, suggesting an excellent hydrolysis effect on starchy raw materials. The hydrolysis degrees of mackerel heads and soybeans by protease were 43.93 ± 2.19% and 26.38 ± 1.72%, respectively, which suggested that the protease from B. velezensis D6 preferentially hydrolyzed animal-based protein., Conclusion: This is a systematic study on the co-production of α-amylase and protease by B. velezensis D6, which is crucial in widening the understanding of this species co-producing multi-enzymes and in exploring its potential application. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Background: 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., Results: 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 (K q = 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., Conclusion: 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., (© 2024 Society of Chemical Industry.)

This study was to investigate the mechanism of the action of epigallocatechin gallate (EGCG) on α-amylase in the ternary simulated system and explore the changes in enzyme structure during the digestion process. Enzymatic kinetics, fluorescence spectroscopy, surface hydrophobicity, fluorescence microscopy, and molecular docking were used to compare (in the presence and absence of EGCG) the structural changes of α-amylase and α-amylase-starch complex, as well as the binding characteristics among EGCG and the α-amylase and starch. The results showed that EGCG had a significant inhibitory effect on α-amylase, and it exhibited a coexistence of competitive and anti-competition inhibition type, and predominantly competitive inhibition. In the ternary and binary systems, the inhibitory mechanisms of EGCG on α-amylase were distinct. In the ternary system, EGCG preferably bound to α-amylase to form α-amylase-EGCG binary complexes rather than α-amylase-starch-EGCG ternary complexes, and altered the structure of α-amylase, leading to unfolding of the enzyme's secondary structure and exposing more non-catalytic site aromatic amino acids., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Cheesemaking with camel milk (CM) presents unique challenges and additional health benefits. This study involved preparing low-fat Cheddar cheese (LFCC) by blending bovine milk (BM) with varying levels of CM. Control cheese was made exclusively with BM. After 180 days of ripening, LFCC samples underwent in vitro digestion to determine antioxidant capacities, α-amylase and α-glucosidase inhibition, and angiotensin-converting enzyme inhibition. The peptide profile of LFCC treatments was analyzed using liquid chromatography-quadrupole-time of flight-mass spectrometry. Antioxidant and biological activities were influenced by BM-CM blends and digestion. At days 120 and 180, the number of αs1-casein-derived peptides increased in all samples except for LFCC made with 15% CM. Generally, 88 peptides exhibited ACE inhibition activity after 120 days of ripening, increasing to 114 by day 180. These findings suggest that ripening time positively affects the health-promoting aspects of functional cheese products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Kafirin in sorghum inhibits starch digestion and exhibits antioxidant properties, however its potential in food industry remains unexplored. Therefore, the study was aimed to explore and improve the potential of kafirin as natural carbohydrate blocker using papain (6 NFU/mL) and/or infrared treatment (220 °C/3 min). Results indicated that the combined treatment, PIR (infrared + papain) is the most efficient treatment to modify kafirin. PIR generated a new ∼37 kDa high molecular weight moiety in kafirin with a crystal size of 157.44 Å. All samples showed superior antioxidant activity post-treatments, with PIR exhibiting highest scavenging activity from 31.09 to 82.97%, 15.09 to 42.82%, and 25.92 to 38.58% for DPPH, FRAP, and ABTS, respectively. PIR-modified kafirin limited malondialdehyde production, and increased α-amylase and α-glucosidase inhibition. Incorporation of 7.5% kafirin in corn starch increased resistant starch from 5.09 to 21.04% after cooking, which suggests potential of kafirin in development of diabetic-friendly food formulations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Euryales Semen (ES) is a highly nutritious food with low digestibility, which is closely associated with its endogenous phenolic compounds. In this study, five phenolic compounds (naringenin, isoquercitrin, gallic acid, epicatechin and quercetin) with high concentrations in ES were selected to prepare starch-polyphenol complexes. Subsequently, the effects of endogenous polyphenols on the structure, physicochemical properties and digestion characteristics of ES starch were studied using multiple techniques. The addition of phenolic compounds markedly reduced the in vitro digestibility, swelling power, gelatinization enthalpy, while increased the solubility of ES starch. Fourier-transform infrared spectroscopy and X-ray diffraction analysis showed that phenolic compounds interacted with the starch through non-covalent bonds. Five phenolic compounds inhibited α-amylase activity through a mixed competitive inhibition mechanism, with the inhibition potency ranked as follows: quercetin > epicatechin > gallic acid > isoquercitrin > naringenin. The spectroscopic analysis and molecular dynamics simulations confirmed that five phenolic compounds interacted with the amino acid residues of α-amylase through hydrogen bonding and hydrophobic interactions, caused α-amylase static fluorescence quenching, and altered its conformation and microenvironment. This study provides a better understanding of the interaction mechanisms between ES starch and polyphenols, and supports the development of ES as a food that lowers sugar levels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In search of novel antidiabetic agents, we synthesized a new series of chalcones with benzimidazole scaffolds by an efficient 'one-pot' nitro reductive cyclization method and evaluated their α-glucosidase and α-amylase inhibition studies. The 'one-pot' nitro reductive cyclization method offered a simple route for the preparation of benzimidazoles with excellent yield and higher purity compared to the other conventional acid- or base-catalyzed cyclization methods. 1 H, 13 C NMR, IR, and mass spectrum data were used to characterize the compounds. Single-crystal XRD data confirmed the 3D structure of compound 7c, which was crystalized in the P1¯ space group of the triclinic crystal system. Hirshfeld surface analysis validates the presence of O-H..O, O-H…N, and C-H…O intermolecular hydrogen bonds. From the DFT calculations, the energy gap between the frontier molecular orbitals in 7c was found to be 3.791 eV. From the series, compound 7l emerged as a potent antidiabetic agent with IC 50 = 22.45 ± 0.36 µg/mL and 20.47 ± 0.60 µg/mL against α-glucosidase and α-amylase enzymes, respectively. The in silico molecular docking studies revealed that compound 7l has strong binding interactions with α-glucosidase and α-amylase proteins. Molecular dynamics studies also revealed the stability of compound 7l with α-glucosidase and α-amylase proteins.

As the concept of precision nutrition has been gradually popularized in recent years, the relationship between the structure of starch-polyphenol complexes with significant health effects and their nutritional functions has been progressively investigated. In this study, G50 high-amylose maize starch with different molecular weights was first prepared by pullulanase and α-amylase, and their effects on the structural formation, digestion properties, and release behaviors of the starch-resveratrol (RA) complex were discussed. The results confirmed that enzyme-treated starch could enhance intermolecular hydrogen bonding and hydrophobic interactions between starch and RA in a high-pressure homogeneous (HPH) environment, forming stable single-helix and V-type crystalline structures while reducing the B-type crystalline structures. Meanwhile, the in vitro experiment showed that when the RA addition was 3%, the resistant starch content of the starch-RA complex could reach 60.3%, and its RA colonic transport rate could reach more than 97%. Interestingly, the starch-RA complex with a relatively higher V-type crystalline structure content contributed to the production of short-chain fatty acids (SCFAs), especially butyrate, and it might be effective in carbohydrate metabolism and immunometabolism by promoting the functions of Phascolarctobacteriu and Alistipes . These findings provide new ideas for the design of the nutritional functions of RS.

Microbial amylases should essentially remain active at higher temperatures, and in the alkaline pH and a range of surfactants to be suitable as detergent additives. In the present study, a thermophilic amylase producing bacterium, Bacillus licheniformis UDS-5 was isolated from Unai hot water spring in Gujarat, India. It was identified as a potent amylase producer during starch plate-based screening process. Therefore, the physicochemical parameters influencing amylase production were optimized using Plackett-Burman design and Central Composite Design. The amylase was purified through ammonium sulfate precipitation, size exclusion and ion exchange chromatography, achieving the purification fold and yield to be 9.2 and 40.6%, respectively. The enzyme displayed robust stability and activity across a wide range of temperatures and pHs, with an increased half-life and reduced deactivation rate constant. The amylase exhibited optimal catalysis at 70 °C and pH 8. The kinetic studies revealed Km and Vmax values of 0.58 mg/mL and 2528 μmol/mL/min, respectively. Besides, the purified amylase displayed stability in the presence of various metal ions, surfactants, and chelators suggesting its potential for industrial applications, particularly in the detergent industry. Moreover, detergent application studies demonstrated its efficacy in enhancing washing performance. A comparative profile on washing efficiency of the studied amylase and the commercial amylase with various detergents pointed towards its possible future use as a detergent additive., Competing Interests: Declarations Conflict of interest The authors declare no conflict of interest. Research involving human participants and/or animals This article does not contain any studies with human participants and/or animals performed by any of the authors., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Postprandial hyperglycemia, typical manifestation of Type 2 Diabetes Mellitus (T2DM), is associated with notable global morbidity and mortality. Preventing the advancement of this condition by delaying the rate of glucose absorption through inhibition of α-amylase and α-glucosidase enzymatic activities is of utmost importance. Finding a safe antidiabetic drug is essential since those that are currently on the market have drawbacks like unpleasant side effects. The current study utilized computer-aided drug design (CADD), as a quick and affordable method to find a substitute drug template that can be used to control postprandial hyperglycemia by modulating the activity of α-amylase and α-glucosidase enzymes. The Natural Products Activity and Species database (NPASS) (30,926 compounds) was screened in silico, with a focus on evaluating drug-likeness, toxicity profiles and ability to bind on a target protein. Two molecules NPC204580 (Chrotacumine C) and NPC137813 (1-O-(2-Methoxy-4-Acetylphenyl)-6-O-(E-Cinnamoyl)-Beta-D-Glucopyranoside) were identified as potential dual inhibitors for α-amylase and α-glucosidase with free binding energies of -14.46 kcal/mol and -12.58 kcal/mol for α-amylase, and -8.42 kcal/mol and -8.76 kcal/mol for α-glucosidase, respectively. The molecules showed ionic, H-bonding and hydrophobic interactions with critical amino acid residues of both enzymes. Moreover, 100 ns molecular dynamic simulations showed that both molecules are stable on the receptors' active sites based on root mean square deviation (RMSD), root mean square fluctuation (RMSF), and the Generalized Born surface area (GBSA) energy calculated. The two compounds are thus promising therapeutic agents for T2DM that merit further investigation due to their excellent binding energies, encouraging pharmacokinetics, toxicity profiles, and stability as demonstrated in simulated studies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ndarawit et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

A key strategy to mitigate postprandial hyperglycemia involves inhibiting α-amylases, which commence the starch digestion process in the gut. This study examined the inhibitory effects of resveratrol and stilbenoid tetramers, vaticanol B, (-)-hopeaphenol, and vatalbinoside A on human salivary and pancreatic α-amylases experimentally and through molecular docking studies. Vaticanol B demonstrated the most potent inhibition with IC 50 values of 5.3 ± 0.3 μM for salivary and 6.1 ± 0.5 μM for pancreatic α-amylase (compared to acarbose with IC 50 values of 1.2 ± 0.1 μM and 0.5 ± 0.0 μM, respectively). Kinetic analysis suggested a competitive inhibition mode for vaticanol B. Resveratrol and vatalbinoside A were poor inhibitors of human α-amylases, while (-)-hopeaphenol exhibited moderate inhibition. Molecular docking supported the inhibition data, and several aspects of the structural configurations explained the stronger inhibition exerted by vaticanol B. Overall, vaticanol B shows promise as a natural alternative to acarbose for inhibiting α-amylase.

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., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Asghar Taheri-Kafrani reports financial support was provided by University of Isfahan. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Porous starch granules (PSGs) are promising biomaterials for the encapsulation, protection, and delivery of bioactive ingredients. In this study, a lipase treatment was first used to generate pores in native starch granules, and then α-amylase was used to enlarge these pores. Electron and fluorescence microscopy analysis showed that the lipase treatment exposed the starch molecules located below the lipid-rich regions on the starch granule surfaces, which increased the swelling of the granules in aqueous solutions. Moreover, lipase treatment caused the surrounding areas to become more loosely packed, which facilitated subsequent starch hydrolysis and the formation of large internal cavities. Fourier transform infrared spectroscopy, X-ray diffraction, and thermogravimetric analyses provided further insights, these methods showed that the short-range order, long-range order, and thermal stability of the PSGs was enhanced by the sequential lipase-amylase modification. PSGs were highly resistant to amylase digestion and had strong adsorption capacity to hydrophobic and hydrophilic substances. This study shows that a combined lipase-amylase treatment can be used to fabricate PSGs, which may have health benefits due to their low digestibility and ability to encapsulate bioactive agents. These PSGs may therefore be suitable for application in the functional food, supplement, personal care, and pharmaceutical industries.

A novel α-amylase gene (BAA) from Bacillus amyloliquefaciens was cloned into Lactococcus lactis , designing two recombinant α-amylases to facilitate extracellular secretion. Following optimizing the expression conditions, the highest yield of BAA (88.12 mmol/L) was achieved upon 36 h induction and 5 ng/mL nisin concentration. Determining the enzymatic properties of BAA revealed its poor stability and activity at high temperatures, hindering its widespread application. Therefore, we used computer-aided design to generate a mutant, S275L, which exhibited significantly improved activity and thermostability: an 18.7% increase in enzymatic activity (3767.38 U/mg), a 10 °C increase in optimal temperature, and a 49.2% improvement in stability at 60 °C. Molecular dynamics simulations and force analysis confirmed these enhancements. Finally, the mutant S275L's potential was further analyzed for starch hydrolysis on poultry feed. Therefore, the mutant S275L holds promising as an enzyme agent for enhanced feed digestibility and quality.

The escalating issue of obesity and related health conditions, including diabetes mellitus and coronary heart disease, necessitates the development of digestive enzyme inhibitors. Targeting pancreatic lipase to inhibit fatty acid generation to attain reduced lipid absorption is a promising approach for identifying effective agents. Orlistat, the only clinically approved pancreatic lipase inhibitor, is associated with gastrointestinal side effects like oily stools, spotting, and flatulence. Concurrently, α-amylase inhibitors can mitigate rapid spikes in blood sugar levels. With the growing significance of natural products in pharmaceutical research, cinnamaldehyde has been recognized as a potential enzyme inhibitor. This study explores the effects of cinnamaldehyde-derived hydrazone Schiff bases on lipase and α-amylase through in vitro experiments. The synthesized compounds exhibited IC 50 value for lipase in the same range ((0.41-12.58) × 10 -8  M) as exhibited by the commercial drug orlistat (5.76 × 10 -8  M). However, for α-amylase inhibition, the compounds (IC 50 value = 0.37-9.54 × 10 -9  M) were found effective inhibitors of acarbose (IC 50 value = 6.76 × 10 -6  M) and curcumin (IC 50 value = 467 × 10 -9  M). In silico DFT studies, molecular docking, drug likeliness, bioactivity score, ADME, and toxicity are also investigated for these compounds., Competing Interests: Declaration of competing interest The Authors declare no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Exploring efficient immobilized carrier for α-Amylase (α-Amy) to enhance its thermostability has significant influence in starch related industry. Here, hollow ZIF-8 (HZIF-8) with amino groups coated magnetic phase change microcapsules (PCM) was designed for covalent immobilization of α-Amy. Magnetic PCM consisting n-docosane core and SiO 2 /Fe 3 O 4 hybrid shell were firstly synthesized. Then, HZIF-8 shell with amino groups was coated and α-Amy was subsequently immobilized through covalent cross-linking strategy. The morphology, chemical structure and magnetic property of PCM@HZIF-8@α-Amy (PCMHA) were comprehensively characterized. Moreover, heat control property of PCMHA was studied with encapsulation efficiency and thermal energy-storage efficiency of 50.55 % and 50.59 %, respectively. Catalytic activity of immobilized α-Amy was fully investigated with K m and V max of 2.773 mg/mL and 1.853 μmol/mL·min, respectively. From reusability and storage stability study, immobilized α-Amy not only maintained rather high activity after 9 cycles' reuse, but also exhibited good activity under high salt ion condition after 7 days., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)