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Genes Jun 2024Starch degradation provides energy and signaling molecules for plant growth, development, defense, and stress response. α-amylase () is one of the most important...
Starch degradation provides energy and signaling molecules for plant growth, development, defense, and stress response. α-amylase () is one of the most important enzymes in this process. Potato tubers are rich in starch, and the hydrolysis of starch into sugar negatively impacts the frying quality of potato. Despite its importance, the gene family has not been fully explored in potatoes. Here, we performed a detailed analysis of the gene family to determine its role in potato. Twenty genes were identified across the potato genome and were divided into three subgroups. The promoters of genes contained an array of -acting elements involved in growth and development, phytohormone signaling, and stress and defense responses. , , , and were specifically expressed in mature tubers. Different gene family members tended to be upregulated in response to β-aminobutyric acid (BABA), (), benzothiadiazole (BTH), heat, salt, and drought stress. In addition, different gene family members tended to be responsive to abscisic acid (ABA), indole-3-acetic acid (IAA), gibberellic acid (GA3), and 6-benzylaminopurine (BAP) treatment. These results suggest that gene family members may be involved in starch and sugar metabolism, defense, stress response, and phytohormone signaling. The results of this study may be applicable to other starchy crops and lay a foundation for further research on the functions and regulatory mechanisms of genes.
Topics: Solanum tuberosum; Gene Expression Regulation, Plant; Plant Proteins; alpha-Amylases; Multigene Family; Stress, Physiological; Plant Growth Regulators; Gene Expression Profiling; Genome, Plant; Plant Tubers; Promoter Regions, Genetic
PubMed: 38927729
DOI: 10.3390/genes15060793 -
Marine Drugs May 2024In this research, the chemical compositions of various extracts obtained from , a type of green seaweed collected from the Nador lagoon in the northern region of...
In this research, the chemical compositions of various extracts obtained from , a type of green seaweed collected from the Nador lagoon in the northern region of Morocco, were compared. Their antioxidant and anti-diabetic properties were also studied. Using GC-MS technology, the fatty acid content of the samples was analyzed, revealing that palmitic acid, eicosenoic acid, and linoleic acid were the most abundant unsaturated fatty acids present in all samples. The HPLC analysis indicated that sinapic acid, naringin, rutin, quercetin, cinnamic acid, salicylic acid, apigenin, flavone, and flavanone were the most prevalent phenolic compounds. The aqueous extract obtained by maceration showed high levels of polyphenols and flavonoids, with values of 379.67 ± 0.09 mg GAE/g and 212.11 ± 0.11 mg QE/g, respectively. This extract also exhibited an impressive ability to scavenge DPPH radicals, as indicated by its IC value of 0.095 ± 0.12 mg/mL. Additionally, the methanolic extract obtained using the Soxhlet method demonstrated antioxidant properties by preventing β-carotene discoloration, with an IC of 0.087 ± 0.14 mg/mL. Results from in-vitro studies showed that extracts from were able to significantly inhibit the enzymatic activity of α-amylase and α-glucosidase. Among the various extracts, methanolic extract (S) has been identified as the most potent inhibitor, exhibiting a statistically similar effect to that of acarbose. Furthermore, molecular docking models were used to evaluate the interaction between the primary phytochemicals found in these extracts and the human pancreatic α-amylase and α-glucosidase enzymes. These findings suggest that extracts contain bioactive substances that are capable of reducing enzyme activity more effectively than the commercially available drug, acarbose.
Topics: Hypoglycemic Agents; Antioxidants; Ulva; Phytochemicals; Plant Extracts; Glycoside Hydrolase Inhibitors; alpha-Amylases; alpha-Glucosidases; Molecular Docking Simulation; Morocco; Humans; Chromatography, High Pressure Liquid; Polyphenols; Flavonoids; Edible Seaweeds
PubMed: 38921551
DOI: 10.3390/md22060240 -
Anais Da Academia Brasileira de Ciencias 2024In this study, the Box-Behnken experimental planning was used to optimize the extraction of polysaccharides from the cell wall of Rhizopus microspore var. oligosporus,...
In this study, the Box-Behnken experimental planning was used to optimize the extraction of polysaccharides from the cell wall of Rhizopus microspore var. oligosporus, with analysis of the quantitative effects of parameters pH, temperature and extraction time for polysaccharide yield. The optimal conditions for extraction were determined by the regression equation and evaluation of the response surface graphs, which indicated: pH 13, temperature of 120ºC and time of 60 min, with maximum yield around 18.5%. Fourier transform infrared spectroscopy analysis indicated typical polysaccharide signals. Nuclear magnetic resonance spectroscopy and monosaccharide analysis indicated a β(1,3) β(1,6) glucogalactan. The polysaccharide exhibited an average molecular weight of 120 kDa and a polymerization degree of 741. Antioxidant assays in vitro revealed the potential of polysaccharide in elimination of ABTS+ radical and hydroxyl radicals. EC50 values for free radical elimination were 7.69 and 17.8 mg/mL, for ABTS+ and hydroxyls, respectively. The polysaccharides showed potential for α-amylase inhibition with an EC50 of 1.66 mg/mL. The results suggest that β(1,3) β(1,6) glucogalactan from Rhizopus microsporus var. oligosporus can be used in biotechnological applications.
Topics: Antioxidants; Rhizopus; alpha-Amylases; Spectroscopy, Fourier Transform Infrared; Galactans; Magnetic Resonance Spectroscopy; Enzyme Inhibitors; Hydrogen-Ion Concentration
PubMed: 38896737
DOI: 10.1590/0001-3765202420230073 -
Molecules (Basel, Switzerland) May 2024Alzheimer's disease (AD) and diabetes are non-communicable diseases with global impacts. Inhibitors of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are...
Alzheimer's disease (AD) and diabetes are non-communicable diseases with global impacts. Inhibitors of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are suitable therapies for AD, while α-amylase and α-glucosidase inhibitors are employed as antidiabetic agents. Compounds were isolated from the medicinal plant and evaluated for their AChE, BChE, α-amylase, and α-glucosidase inhibitions. From H and C NMR data, the compounds were identified as 3,3'-di-O-methyl ellagic acid (), 3,3',4'-tri-O-methyl ellagic acid-4-O-β-D-xylopyranoside (), 3,3',4'-tri-O-methyl ellagic acid-4-O-β-D-glucopyranoside (), 3,3'-di-O-methyl ellagic acid-4-O-β-D-glucopyranoside (), myricetin-3-O-rhamnoside (), shikimic acid (), arjungenin (), terminolic acid (), 24-deoxysericoside (), arjunglucoside I (), and chebuloside II (). The derivatives of ellagic acid (-) showed moderate to good inhibition of cholinesterases, with the most potent being 3,3'-di-O-methyl ellagic acid, with IC values of 46.77 ± 0.90 µg/mL and 50.48 ± 1.10 µg/mL against AChE and BChE, respectively. The compounds exhibited potential inhibition of α-amylase and α-glucosidase, especially the phenolic compounds (-). Myricetin-3-O-rhamnoside had the highest α-amylase inhibition with an IC value of 65.17 ± 0.43 µg/mL compared to acarbose with an IC value of 32.25 ± 0.36 µg/mL. Two compounds, 3,3'-di-O-methyl ellagic acid (IC = 74.18 ± 0.29 µg/mL) and myricetin-3-O-rhamnoside (IC = 69.02 ± 0.65 µg/mL), were more active than the standard acarbose (IC = 87.70 ± 0.68 µg/mL) in the α-glucosidase assay. For α-glucosidase and α-amylase, the molecular docking results for reveal that these compounds may fit well into the binding sites of the target enzymes, establishing stable complexes with negative binding energies in the range of -4.03 to -10.20 kcalmol. Though not all the compounds showed binding affinities with cholinesterases, some had negative binding energies, indicating that the inhibition was thermodynamically favorable.
Topics: Molecular Docking Simulation; Cholinesterase Inhibitors; Hypoglycemic Agents; Plant Extracts; alpha-Amylases; Acetylcholinesterase; Terminalia; Humans; Butyrylcholinesterase; alpha-Glucosidases; Glycoside Hydrolase Inhibitors; Molecular Structure
PubMed: 38893333
DOI: 10.3390/molecules29112456 -
Plants (Basel, Switzerland) Jun 2024Alpha-amylases are crucial hydrolase enzymes which have been widely used in food, feed, fermentation, and pharmaceutical industries. Methods for low-cost production of...
Alpha-amylases are crucial hydrolase enzymes which have been widely used in food, feed, fermentation, and pharmaceutical industries. Methods for low-cost production of α-amylases are highly desirable. Soybean seed, functioning as a bioreactor, offers an excellent platform for the mass production of recombinant proteins for its ability to synthesize substantial quantities of proteins. In this study, we generated and characterized transgenic soybeans expressing the α-amylase AmyS from . The α-amylase expression cassettes were constructed for seed specific expression by utilizing the promoters of three different soybean storage peptides and transformed into soybean via -mediated transformation. The event with the highest amylase activity reached 601 U/mg of seed flour (one unit is defined as the amount of enzyme that generates 1 micromole reducing ends per min from starch at 65 °C in pH 5.5 sodium acetate buffer). The optimum pH, optimum temperature, and the enzymatic kinetics of the soybean expressed enzyme are similar to that of the expressed enzyme. However, the soybean expressed α-amylase is glycosylated, exhibiting enhanced thermostability and storage stability. Soybean AmyS retains over 80% activity after 100 min at 75 °C, and the transgenic seeds exhibit no significant activity loss after one year of storage at room temperature. The accumulated AmyS in the transgenic seeds represents approximately 15% of the total seed protein, or about 4% of the dry seed weight. The specific activity of the transgenic soybean seed flour is comparable to many commercial α-amylase enzyme products in current markets, suggesting that the soybean flour may be directly used for various applications without the need for extraction and purification.
PubMed: 38891347
DOI: 10.3390/plants13111539 -
Inhibition of human starch digesting enzymes and intestinal glucose transport by walnut polyphenols.Food Research International (Ottawa,... Aug 2024One approach to controlling type 2 diabetes (T2D) is to lower postprandialglucose spikesby slowing down the digestion of carbohydrates and the absorption of glucose in...
One approach to controlling type 2 diabetes (T2D) is to lower postprandialglucose spikesby slowing down the digestion of carbohydrates and the absorption of glucose in the small intestine. The consumption of walnuts is associated with a reduced risk of chronic diseases such as T2D, suggested to be partly due to the high content of (poly)phenols. This study evaluated, for the first time, the inhibitory effect of a (poly)phenol-rich walnut extract on human carbohydrate digesting enzymes (salivary and pancreatic α-amylases, brush border sucrase-isomaltase) and on glucose transport across fully differentiated human intestinal Caco-2/TC7 monolayers. The walnut extract was rich in multiple (poly)phenols (70 % w/w) as analysed by Folin-Ciocalteau and by LCMS. It exhibited potent inhibition of both human salivary (IC: 32.2 ± 2.5 µg walnut (poly)phenols (WP)/mL) and pancreatic (IC: 56.7 ± 1.7 µg WP/mL) α-amylases, with weaker effects on human sucrase (IC: 990 ± 20 µg WP/mL), maltase (IC: 1300 ± 80 µg WP/mL), and isomaltase (IC: 830 ± 60 µg WP/mL) activities. Selected individual walnut (poly)phenols inhibited human salivary α-amylase in the order: 1,3,4,6-tetragalloylglucose > ellagic acid pentoside > 1,2,6-tri-O-galloyl-β-D-glucopyranose, with no inhibition by ellagic acid, gallic acid and 4-O-methylgallic acid. The (poly)phenol-rich walnut extract also attenuated (up to 59 %) the transfer of 2-deoxy-D-glucose across differentiated Caco-2/TC7 cell monolayers. This is the first report on the effect of (poly)phenol-rich extracts from any commonly-consumed nut kernel on any human starch-digesting enzyme, and suggests a mechanism through which walnut consumption may lower postprandial glucose spikes and contribute to their proposed health benefits.
Topics: Humans; Polyphenols; Juglans; Caco-2 Cells; Glucose; Plant Extracts; Digestion; Nuts; Starch; alpha-Amylases; Biological Transport; Sucrase-Isomaltase Complex
PubMed: 38876610
DOI: 10.1016/j.foodres.2024.114572 -
AMB Express Jun 2024α-Amylase plays a crucial role in the industrial degradation of starch. The genus Jeotgalibacillus of the underexplored marine bacteria family Caryophanaceae has not...
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 D5 (= DSM28777 = KCTC33550). 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 CaCl 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.
PubMed: 38874807
DOI: 10.1186/s13568-024-01722-3 -
Journal of Enzyme Inhibition and... Dec 2024Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease mainly caused by insulin resistance, which can lead to a series of complications such as cardiovascular...
Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease mainly caused by insulin resistance, which can lead to a series of complications such as cardiovascular disease, retinopathy, and its typical clinical symptom is hyperglycaemia. Glucosidase inhibitors, including Acarbose, Miglitol, are commonly used in the clinical treatment of hypoglycaemia. In addition, Protein tyrosine phosphatase 1B (PTP1B) is also an important promising target for the treatment of T2DM. is a well-known oriental traditional medicinal herbal plant, and has many beneficial effects on glucose and lipid metabolism. In the present study, three new and nine known dammarane triterpenoids isolated from , and their structures were elucidated by spectroscopic methods including HR-ESI-MS,H and C NMR and X-ray crystallography. All these compounds were evaluated for inhibitory activity against α-glucosidase, α-amylase and PTP1B. The results suggested that compounds ∼ were potential antidiabetic agents with significantly inhibition activity against PTP1B in a dose-dependent manner.
Topics: Protein Tyrosine Phosphatase, Non-Receptor Type 1; Gynostemma; Enzyme Inhibitors; Molecular Structure; Dose-Response Relationship, Drug; Structure-Activity Relationship; alpha-Glucosidases; Humans; alpha-Amylases; Hypoglycemic Agents; Models, Molecular; Crystallography, X-Ray; Triterpenes; Glycoside Hydrolase Inhibitors
PubMed: 38873930
DOI: 10.1080/14756366.2024.2360063 -
BMC Complementary Medicine and Therapies Jun 2024Diabetes affects 75% of people in low-income countries, where conventional drugs like metformin are available, but newer drugs like alpha-glucosidase inhibitors are not...
BACKGROUND
Diabetes affects 75% of people in low-income countries, where conventional drugs like metformin are available, but newer drugs like alpha-glucosidase inhibitors are not accessible to most Southern African patients.
AIM
To evaluate the α-glucosidase and α-amylase inhibitory activities of fractionated aqueous extracts of Kigelia africana fruit (KAFE) and their phytochemical fingerprints using gas chromatography-mass spectrometry (GC-MS).
MATERIALS AND METHODS
We studied K. africana fruit fractions' inhibitory effects on alpha-glucosidase and alpha-amylase using bioassay-guided fractionation, and analyzed their phytochemical profiles with GC-MS.
KEY FINDINGS
Both the aqueous extract and ethyl acetate fraction of the aqueous extract exhibited a low dose-dependent inhibition of alpha-amylase activity (p < 0.0001). At a concentration of 500 μg/mL, the aqueous extract caused an alpha-glucosidase inhibition of 64.10 ± 2.7%, with an estimated IC50 of 193.7 μg/mL, while the ethyl acetate fraction had an inhibition of 89.82 ± 0.8% and an estimated IC50 of 10.41 μg/mL. The subfraction G, which had the highest alpha-glucosidase inhibitory activity at 85.10 ± 0.7%, had significantly lower activity than the ethyl acetate fraction. The most bioactive fraction was found to contain 11"(2-cyclopenten-1-yl) undecanoic acid, ( +)- and cyclopentane undecanoic acid as well as the indole alkaloids Akuammilan-17-ol-10-methoxy, N-nitroso-2-methyl-oxazolidine and epoxide Oxirane2.2″ -(1.4-butanediyl) bis-.
CONCLUSION
The K. africana fruit fraction demonstrated significant alpha-glucosidase inhibitory activity, while its alpha-amylase inhibitory activity was limited. This study suggests a potential natural alpha-glucosidase inhibitor and phytocompounds that could serve as leads for developing antidiabetic agents.
Topics: Glycoside Hydrolase Inhibitors; Plant Extracts; Fruit; alpha-Glucosidases; alpha-Amylases; Gas Chromatography-Mass Spectrometry; Humans; Phytochemicals
PubMed: 38867199
DOI: 10.1186/s12906-024-04510-5 -
Scientific Reports Jun 2024In the field of biotechnology, the utilization of agro-industrial waste for generating high-value products, such as microbial biomass and enzymes, holds significant...
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. 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) while exhibiting comparatively lower activity towards cyclodextrins and amylose (≤ 200.2 ± 16.2 U mg). 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, 5,5'-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 CaCl (2 mM) even led to a slight activation of the recombinant enzyme (1.4 times). The Michaelis constant (K) and maximum reaction rate (V), were determined using soluble starch as a substrate, yielding values of 1.2 ± 0.19 mg mL and 1580.3 ± 183.7 μmol mg protein min, 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.
Topics: alpha-Amylases; Whey; Anoxybacillus; Detergents; Hydrogen-Ion Concentration; Enzyme Stability; Recombinant Proteins; Starch; Temperature
PubMed: 38830978
DOI: 10.1038/s41598-024-63606-7