Salivary Α-amylases Research Articles

Inhibition of Human Salivary and Pancreatic α-Amylase by Resveratrol Oligomers.

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 IC50 values of 5.3 ± 0.3 μM for salivary and 6.1 ± 0.5 μM for pancreatic α-amylase (compared to acarbose with IC50 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.

Structure-function relationships in (poly)phenol-enzyme binding: Direct inhibition of human salivary and pancreatic α-amylases

(Poly)phenols inhibit α-amylase by directly binding to the enzyme and/or by forming starch-polyphenol complexes. Conventional methods using starch as the substrate measure inhibition from both mechanisms, whereas the use of shorter oligosaccharides as substrates exclusively measures the direct interaction of (poly)phenols with the enzyme. In this study, using a chromatography-based method and a short oligosaccharide as the substrate, we investigated the detailed structural prerequisites for the direct inhibition of human salivary and pancreatic α-amylases by over 50 (poly)phenols from the (poly)phenol groups: flavonols, flavones, flavanones, flavan-3-ols, polymethoxyflavones, isoflavones, anthocyanidins and phenolic acids. Despite being structurally very similar (97% sequence homology), human salivary and pancreatic α-amylases were inhibited to different extents by the tested (poly)phenols. The most potent human salivary α-amylase inhibitors were luteolin and pelargonidin, while the methoxylated anthocyanidins, peonidin and petunidin, significantly blocked pancreatic enzyme activity. B-ring methoxylation of anthocyanidins increased inhibition against both human α-amylases while hydroxyl groups at C3 and B3′ acted antagonistically in human salivary inhibition. C4 carbonyl reduction, or the positive charge on the flavonoid structure, was the key structural feature for human pancreatic inhibition. B-ring glycosylation did not affect salivary enzyme inhibition, but increased pancreatic enzyme inhibition when compared to its corresponding aglycone. Overall, our findings indicate that the efficacy of interaction with human α-amylase is mainly influenced by the type and placement of functional groups rather than the number of hydroxyl groups and molecular weight.

Inhibition of human starch digesting enzymes and intestinal glucose transport by walnut polyphenols

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 (IC50: 32.2 ± 2.5 µg walnut (poly)phenols (WP)/mL) and pancreatic (IC50: 56.7 ± 1.7 µg WP/mL) α-amylases, with weaker effects on human sucrase (IC50: 990 ± 20 µg WP/mL), maltase (IC50: 1300 ± 80 µg WP/mL), and isomaltase (IC25: 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.

Antifungal, Antimycobacterial, Protease and α‒Amylase Inhibitory Activities of a Novel Serine Bifunctional Protease Inhibitor from Adenanthera pavonina L. Seeds.

Antifungal resistance poses a significant challenge to disease management, necessitating the development of novel drugs. Antimicrobial peptides offer potential solutions. This study focused on extraction and characterization of peptides from Adenanthera pavonina seeds with activity against Candida species, Mycobacterium tuberculosis, proteases, and α-amylases. Peptides were extracted in phosphate buffer and heated at 90°C for 10 min to create a peptide rich heated fraction (PRHF). After confirming antimicrobial activity and the presence of peptides, the PRHF underwent ion exchange chromatography, yielding retained and non-retained fractions. These fractions were evaluated for antimicrobial activity and cytotoxicity against murine macrophages. The least toxic and most active fraction underwent reversed-phase chromatography, resulting in ten fractions. These fractions were tested for peptides and antimicrobial activity. The most active fraction was rechromatographed on a reversed-phase column, resulting in two fractions that were assessed for antimicrobial activity. The most active fraction revealed a single band of approximately 6 kDa and was tested for inhibitory effects on proteases and α-amylases. Thermal stability experiments were conducted on the 6 kDa peptide at different temperatures followed by reassessment of antifungal activity and circular dichroism. The 6 kDa peptide inhibited yeasts, M. tuberculosis, human salivary and Tenebrio molitor larvae intestine α-amylases, and proteolytic activity from fungal extracts, and thus named ApPI. Remarkably, ApPI retained antifungal activity and conformation after heating and is primarily composed of α-helices. ApPI is a thermally stable serine protease/α-amylase inhibitor from A. pavonina seeds, offering promise as a foundational molecule for innovative therapeutic agents against fungal infections and tuberculosis.

Control of postprandial hyperglycemia by oral administration of the sea anemone mucus-derived α-amylase inhibitor (magnificamide)

Diabetes mellitus is a serious threat to human health in both developed and developing countries. Optimal disease control requires the use of a diet and a combination of several medications, including oral hypoglycemic agents such as α-glucosidase inhibitors. Currently, the arsenal of available drugs is insufficient, which determines the relevance of studying new potent α-amylase inhibitors. We implemented the recombinant production of sea anemone derived α-amylase inhibitor magnificamide in Escherichia coli. Peptide was isolated by a combination of liquid chromatography techniques. Its folding and molecular weight was proved by 1H NMR and mass spectrometry. The Ki value of magnificamide against human pancreatic α-amylase is 3.1 nM according to Morrison equation for tight binding inhibitors. Our study of the thermodynamic characteristics of binding of magnificamide to human salivary and pancreatic α-amylases by isothermal titration calorimetry showed the presence of different binding mechanisms with Kd equal to 0.11 µM and 0.1 nM, respectively. Experiments in mice with streptozotocin-induced diabetes mimicking diabetes mellitus type 1 were used to study the efficiency of magnificamide against postprandial hyperglycemia. It was found that at a dose of 0.005 mg kg–1, magnificamide effectively blocks starch breakdown and prevents the development of postprandial hyperglycemia in T1D mice. Our results demonstrated the therapeutic potential of magnificamide for the control of postprandial hyperglycemia.

Conjugated hypercrosslinked polymers imprinted with 3,5-dinitrosalicylic acid for the fluorescent determination of α-amylase activity

The discovery of a series of coupling reactions between various building blocks has driven the development of porous organic polymers, but the common usage of expensive and air-sensitive organometallic catalysts and complex procedures in harsh syntheses has limited their expansion. A microporous hypercrosslinked polymer (HCP) was synthesized by polymerizing a naphthalene monomer and a 1,4-dimethoxybenzene crosslinker using Friedel–Crafts alkylation over an FeCl3 catalyst and imprinted with 3,5-dinitrosalicylic acid (DNS). The DNS-molecularly-imprinted HCPs (MIHCPs) were characterized as having IUPAC Type I mesoporosity, a specific surface area of 1134 m2 g−1, a monolayer adsorption capacity of 116 cm2 g−1, pore sizes ranging from 5 to 8.5 Å, amorphous frameworks, and distinctive absorption and emission bands by N2 adsorption/desorption analyses, scanning and transmission electron microscopies, and FTIR, UV–Vis, and fluorescence spectrometries. The π-conjugated imprinted framework endowed the MIHCPs with 405-nm fluorescent emission at a 330-nm excitation and dynamic quenching, which was confirmed by changes in fluorescence lifetime and followed a linear Stern-Volmer plot against 1.0–200 μM DNS template molecules under optimized conditions of a pH 7.0 buffer, an MIHCP concentration of 125 μg mL−1, and a 3.0-min equilibration time. The MIHCPs exhibited a high imprinted factor of 8.7 against nonimprinted HCP and a selectivity of 8.63 against reduced DNS, which enabled fluorometric detection of DNS molecules produced by the hydrolysis of starch with microbial, salivary, and pancreatic α-amylases and the subsequent redox incubation with the DNS oxidant. The fluorometric measurement of α-amylase activity was higher in accuracy and precision (RSD: 2.6–2.8% vs. 3.9–4.0%) than conventional UV–Vis spectrometry because the excellent fluorescent sensitivity and imprinting selectivity of the MIHCP probes enabled the use of higher dilution factors with weaker matrix effects.

Characterization of pearl millet oligosaccharides and evaluation of their prebiotic potential

In the present study pearl millet oligosaccharides (PMOs) were extracted at optimized process parameters and further purified by passing through charcoal column and dialysis membrane (500 Da) to obtain a purity >90% with a yield of 4.6% (w/w). The molecular weights of the mixed oligosaccharides (342, 504, 667, 829, 991, 1153, 1316 Da) were confirmed by MALDI-TOF. The non-digestibility assay was performed by acid hydrolysis, salivary and pancreatic α amylases. The results showed a maximum hydrolysis of 0.2516% ± 0.0173, 2.8% ± 0.023 and 3.1% ± 0.11 by gastric acid, salivary and pancreatic α-amylases respectively. A positive prebiotic score, enhanced biofilm formation and capability of producing SCFAs (i.e. acetate, propionate and butyrate) by different strains of lactobacilli in presence of purified oligosaccharides hints towards their prebiotic potential. From the observations we propose that the PMOs can be explored further as nutraceuticals and functional food additives.

Slowly digestible property of highly branched α-limit dextrins produced by 4,6-α-glucanotransferase from Streptococcus thermophilus evaluated in vitro and in vivo

Starch molecules are first degraded to slowly digestible α-limit dextrins (α-LDx) and rapidly hydrolyzable linear malto-oligosaccharides (LMOs) by salivary and pancreatic α-amylases. In this study, we designed a slowly digestible highly branched α-LDx with maximized α-1,6 linkages using 4,6-α-glucanotransferase (4,6-αGT), which creates a short length of α-1,4 side chains with increasing branching points. The results showed that a short length of external chains mainly composed of 1–8 glucosyl units was newly synthesized in different amylose contents of corn starches, and the α-1,6 linkage ratio of branched α-LDx after the chromatographical purification was significantly increased from 4.6% to 22.1%. Both in vitro and in vivo studies confirmed that enzymatically modified α-LDx had improved slowly digestible properties and extended glycemic responses. Therefore, 4,6-αGT treatment enhanced the slowly digestible properties of highly branched α-LDx and promises usefulness as a functional ingredient to attenuate postprandial glucose homeostasis.

Potential anti-diabetic properties of Merlot grape pomace extract: An in vitro, in silico and in vivo study of α-amylase and α-glucosidase inhibition

A practical approach to control glycemia in diabetes is to use plant natural products that delay hydrolysis of complex sugars and promote the diminution of the release of glucosyl units into the blood plasma. Polyphenolics have been described as being effective in inhibiting amylases and α-glucosidases. Grape pomace is an important sub product of the wine industry, still rich in many compounds such as polyphenolics. In this context, the purpose of this study was to search for possible effects of a grape pomace extract on salivary and pancreatic α-amylases and α-glucosidase, as well as on intestinal glucose absorption. The Merlot grape pomace extract (MGPE) was prepared using a hydroalcoholic mixture (40% ethanol + 60% water). In vitro inhibition was quantified using potato starch (for amylases) and maltose (for α-glucosidase) as substrates. In vivo inhibition was evaluated by running starch and maltose tolerance tests in rats with or without administration of MGPE. Ranking of the extract compounds for its affinity to the α-amylases was accomplished by computer simulations using three different programs. Both α-amylases, pancreatic and salivary, were inhibited by the MGPE. No inhibition on α-glucosidase, however, was detected. The IC50 values were 90 ± 10 μg/mL and 143 ± 15 μg/mL for salivary and pancreatic amylases, respectively. Kinetically this inhibition showed a complex pattern, with multiple binding of the extract constituents to the enzymes. Furthermore, the in silico docking simulations indicated that several phenolic substances, e.g., peonidin-3-O-acetylglucoside, quercetin-3-O-glucuronide and isorhamnetin-3-O-glucoside, besides catechin, were the most likely polyphenols responsible for the α-amylase inhibition caused by MGPE. The hyperglycemic burst, an usual phenomenon that follows starch administration, was substantially inhibited by the MGPE. Our results suggest that the MGPE can be adequate for maintaining normal blood levels after food ingestion.

Highly branched α-limit dextrins attenuate the glycemic response and stimulate the secretion of satiety hormone peptide YY

Starch molecules are hydrolyzed to linear malto-oligosaccharides and branched α-limit dextrins (α-LDx) by salivary and pancreatic α-amylases in the gastrointestinal tract. In this study, we investigated the physiological responses to chromatographically purified branched α-LDx, which are slowly hydrolyzed owing to their high α-1,6 linkage ratios. Our results demonstrate that the digestion of branched α-LDx by mammalian α-glucosidases is slower than that of malto-oligosaccharides in vitro. In vivo experiments using mice showed that the glucose spike at the initial stage of α-LDx hydrolysis was significantly (p < 0.05) attenuated owing to an extended glycemic response. Branched α-LDx stimulated the secretion of peptide YY, which controls the satiety level and rate of gastric emptying by regulating the hypothalamus activity. In conclusion, the slow digestibility of branched α-LDx decelerated glucogenesis in vitro and in vivo; this phenomenon may result in beneficial physiological effects for regulate type 2 diabetes, obesity, and related chronic diseases by controlling post-prandial glucose homeostasis.

A digestive tract expressing α-amylase influences the adult lifespan of Pteromalus puparum revealed through RNAi and rescue analyses.

Midgut and salivary gland α-amylases are digestive enzymes required for the development of insects and have been investigated in some insect species. However, α-amylases in the endoparasitioid wasps have not been reported. Pteromalus puparum (Hymenoptera: Pteromalidae) is a dominant endoparasitioid wasp that parasitizes many butterfly species, including the Brassicaceae pest Pieris rapae (Lepidoptera: Pieridae). Here, we studied the characteristics and functions of three α-amylases in P. puparum. We cloned three genes encoding α-amylases in P. puparum, PpAmy1, PpAmy2 and PpAmy3. The full length of the PpAmy1 cDNA is 1872 bp, encoding 496 amino acids, the PpAmy2 cDNA is 1863 bp long, encoding 518 amino acids, and PpAmy3 cDNA consists of 1802 bp encoding 521 amino acids. PpAmys are highly similar in amino acid sequences, but they have separate tissue distributions. Phylogenetic results show that gene duplications may occur between PpAmy2 and PpAmy3. PpAmy1 and PpAmy3 are most highly expressed in the digestive tract and the venom apparatus, respectively, while PpAmy2 is broadly expressed in all tissues. We report that PpAmy1 acts in the digestive tract, where it influences lifespan as demonstrated using RNAi and α-amylase rescue analyses, and there is no significant difference in longevity when PpAmy2 and PpAmy3 are knocked down. PpAmys probably have roles in carbohydrate metabolism of P. puparum and its host/parasitoid relationships. The characterization and functional study of PpAmys lays the foundation for the protection and utilization of parasitoid resources, and the biological control of agricultural pests. © 2019 Society of Chemical Industry.

Oro-gastro-intestinal digestion of starch in white bread, wheat-based and gluten-free pasta: Unveiling the contribution of human salivary α-amylase

Starch is a major determinant of the glycemic responses elicited by our diets, but the exact contribution of the two main amylolytic enzymes (salivary and pancreatic α-amylases) remains a matter of debate. Our aim was to investigate the contribution of the oral, gastric and intestinal phases to the hydrolysis of starch in bread and pasta during dynamic in vitro digestions using DiDGI®. Before its inactivation by the low gastric pH, salivary α-amylase released about 80% of the starch in bread and 30% of that in pasta, hydrolysing over half of it into oligosaccharides. Accordingly, the contribution of pancreatic α-amylase during the intestinal phase was lower for bread than pasta. Our results are well correlated with in vivo data, and demonstrate the importance of salivary α-amylase during oro-gastric processing of starchy foods. This finding is discussed in relation with observations regarding salivary α-amylase from other fields of knowledge.

Inhibition of α-amylases by pentagalloyl glucose: Kinetics, molecular dynamics and consequences for starch absorption

Pentagalloyl-glucose (PGG) occurs in many plants and possesses several therapeutic properties, including anti-diabetic ones. Based on the latter, the inhibitory action of PGG on α-amylases was investigated. PGG inhibited both the pancreatic and the salivary α-amylases. With starch as the substrate, complexation with the free enzyme form either predominates (pancreatic enzyme) or is the only complexation that is effectively allowed (salivary enzyme; competitive inhibition). This contrasts with a previous report on the salivary enzyme showing non-competitive inhibition when a synthetic substrate was used. PGG was less effective than acarbose as an inhibitor of α-amylases and presented a different binding pattern in terms of the aminoacyl residues involved. Finally, PGG also diminished the hyperglycemic excursion normally caused by starch administration when it was given to rats simultaneously to the polysaccharide. This suggests that PGG or foods and infusions rich in PGG could be helpful as adjuvants for maintaining normal glycemia.

Inhibition of α-Amylases by Condensed and Hydrolysable Tannins: Focus on Kinetics and Hypoglycemic Actions.

The aim of the present study was to compare the in vitro inhibitory effects on the salivary and pancreatic α-amylases and the in vivo hypoglycemic actions of the hydrolysable tannin from Chinese natural gall and the condensed tannin from Acacia mearnsii. The human salivary α-amylase was more strongly inhibited by the hydrolysable than by the condensed tannin, with the concentrations for 50% inhibition (IC50) being 47.0 and 285.4 μM, respectively. The inhibitory capacities of both tannins on the pancreatic α-amylase were also different, with IC50 values being 141.1 μM for the hydrolysable tannin and 248.1 μM for the condensed tannin. The kinetics of the inhibition presented complex patterns in that for both inhibitors more than one molecule can bind simultaneously to either the free enzyme of the substrate-complexed enzyme (parabolic mixed inhibition). Both tannins were able to inhibit the intestinal starch absorption. Inhibition by the hydrolysable tannin was concentration-dependent, with 53% inhibition at the dose of 58.8 μmol/kg and 88% inhibition at the dose of 294 μmol/kg. For the condensed tannin, inhibition was not substantially different for doses between 124.4 μmol/kg (49%) and 620 μmol/kg (57%). It can be concluded that both tannins, but especially the hydrolysable one, could be useful in controlling the postprandial glycemic levels in diabetes.

Development of a workflow for screening and identification of α-amylase inhibitory peptides from food source using an integrated Bioinformatics-phage display approach: Case study – Cumin seed

The main objective of this study was to develop an efficient workflow to discover α-amylase inhibitory peptides from cumin seed. A total of 56 unknown peptides was initially found in the cumin seed protein hydrolysate. They were subjected to 2 different in silico screenings and 6 peptides were shortlisted. The peptides were then subjected to in vitro selection using phage display technique and 3 clones (CSP3, CSP4 and CSP6) showed high affinity in binding α-amylase. These clones were subjected to the inhibitory test and only CSP4 and CSP6 exhibited high inhibitory activity. Therefore, these peptides were chemically synthesized for validation purposes. CSP4 exhibited inhibition of bacterial and human salivary α-amylases with IC50 values of 0.11 and 0.04μmol, respectively, whereas CSP6 was about 0.10 and 0.15μmol, respectively. Results showed that the strength of each protocol has been successfully combined as deemed fit to enhance the α-amylase inhibitor peptide discovery.

Correlation of pasting behaviors with total phenolic compounds and starch digestibility of indigenous pigmented rice grown in upper Northern Thailand

Background: Thailand has one of the most important rice genetic resources with white, light brown, brown, red, and purple rice bran colors. The latter believed to have potential for health benefits due to their phenolic content. Recently researchers have indicated that starch digestive enzymes, including salivary and pancreatic α-amylases and α-glucosidases, can be inhibited by phenolic compounds. Although pasting properties of rice flour are key determinants of quality significantly impacting the final product texture, there is no in-depth study on their correlation with phenolic compound and starch digestibility. Methods: Rice flour from twelve varieties, three from each of five bran colors (white, brown, red, and purple), were evaluated for pasting properties (RVA-3D), total phenolic compounds, amylose content, resistant starch and estimated glycemic index. Simple correlation coefficients were calculated for the relationships between pasting properties (final viscosity, breakdown, setback and pasting temperature) and total phenolic compounds, resistant starch and estimated glycemic index.Results: Within each rice variety, red and purple pigmented flours had higher total phenolic compounds (TPC) and more resistant starch than that of white flours. The TPC and resistant starch content of the flours ranged between 7.83- 47.3 mg/L and 2.44–10.50% respectively, and producing 60-80 of estimated glycemic index. Viscosity behavior showed that pigmented with low amylose rice had lower viscosity temperature than that of pigmented with high amylose rice flour, but higher in peak viscosity. Correlation coefficients of pasting temperature, final viscosity, break down and setback with TCP was observed to be inversely related to glycemic index. However, it was positively correlated to the resistant starch and amylose content. Conclusions: Pigmented rice flour is a better source of TPC and resistant starch which in turn provides low glycemic index. This could help reduce the onset of type 2 diabetes and other related chronic diseases. In addition, this study provides impact of pasting behavior – TPC- resistant starch of rice flour relationships, which have important implication for utilization in food industry.Keywords: total phenolic compounds, amylose content, resistant starch, glycemic index

CARBOHYDRASES IN THE DIGESTIVE SYSTEM OF THE SPINED SOLDIER BUG, Podisus maculiventris (SAY) (HEMIPTERA: PENTATOMIDAE)

The spined soldier bug, Podisus maculiventris, is a generalist predator of insects and has been used in biological control. However, information on the digestion of food in this insect is lacking. Therefore, we have studied the digestive system in P. maculiventris, and further characterized carbohydrases in the digestive tract. The midgut of all developmental stages was composed of anterior, median, and posterior regions. The volumes of the anterior midgut decreased and the median midgut increased in older instars and adults, suggesting a more important role of the median midgut in food digestion. However, carbohydrase activities were predominant in the anterior midgut. In comparing the specific activity of carbohydrases, α-amylase activity was more in the salivary glands (with two distinct activity bands in zymograms), and glucosidase and galactosidase activities were more in the midgut. Salivary α-amylases were detected in the prey hemolymph, demonstrating the role of these enzymes in extra-oral digestion. However, the catalytic efficiency of midgut α-amylase activity was approximately twofold more than that of the salivary gland enzymes, and was more efficient in digesting soluble starch than glycogen. Midgut α-amylases were developmentally regulated, as one isoform was found in first instar compared to three isoforms in fifth instar nymphs. Starvation significantly affected carbohydrase activities in the midgut, and acarbose inhibited α-amylases from both the salivary glands and midgut in vitro and in vivo. The structural diversity and developmental regulation of carbohydrases in the digestive system of P. maculiventris demonstrate the importance of these enzymes in extra-oral and intra-tract digestion, and may explain the capability of the hemipteran to utilize diverse food sources.

Inhibition of salivary and pancreatic α-amylases by a pinhão coat (Araucaria angustifolia) extract rich in condensed tannin

The purpose of the present work was to investigate the possible inhibitory effect of a pinhão coat extract rich in condensed tannin on the activity of α-amylases (human salivary and porcine pancreatic). Experiments with the classic α-amylase inhibitor acarbose and a condensed tannin from Acacia mearnsii were done for comparative purposes. Fourier transform-infrared spectroscopy analysis of the pinhão coat tannin revealed a higher proportion of procyanidins to prodelphinidins when compared to the A. mearnsii tannin. The pinhão coat extract rich in condensed tannin was an effective inhibitor of both human salivary and porcine pancreatic α-amylase. Inhibition was of the mixed S-parabolic I-parabolic type. For the human salivary α-amylase the inhibition constants were 56.88±5.74 and 103.27±11.85μg/L; for the porcine pancreatic α-amylase the inhibition constants were smaller, namely, 20.25±1.97 and 46.79±4.57μg/L. The decreasing potency sequence was: acarbose>pinhão coat extract rich in tannin>A. mearnsii tannin. Similarly to acarbose and the A. mearnsii tannin, the pinhão coat extract was also effective in diminishing the post-prandial glycemic levels in rats after starch administration. The inhibitory properties of the pinhão coat extract indicate that it can be used to suppress postprandial hyperglycemia in diabetic patients.

Enzymatic extraction of beta-glucan from oat bran cereals and oat crackers and optimization of viscosity measurement

The viscosity of the soluble fibre, β-glucan, has been shown to influence its ability to lower serum cholesterol and postprandial blood glucose levels. The impact of various amylases, proteases and lipase on the solubility and resulting viscosity of β-glucan extracted from oat bran cereals with a range of β-glucan concentrations and molecular weights was investigated. Addition of enzymes increased the final viscosity of high molecular weight β-glucan in cereals by facilitating the release of β-glucan from the food matrix. For cereals with partially depolymerized β-glucan, the addition of digestive enzymes decreased the final viscosity by eliminating the contribution of starch and protein to viscosity. Final viscosity varied depending on enzyme combinations including pancreatin, salivary and microbial α-amylases, microbial protease, porcine protease, trypsin and α-chymotrypsin. Addition of lipase did not significantly affect viscosity or solubility of β-glucan extracted from oat crackers. Addition of lichenase showed that β-glucan was the major contributor of viscosity to the system, with negligible interference from other components. The viscosity of the optimized protocol was compared to physiological results previously obtained. The viscosity of β-glucan extracted with pancreatin plus microbial α-amylase (pH 6.9) was predictive of LDL-cholesterol reduction (R2 = 0.847) and glycemic response (R2 = 0.883).

Naturally Occurring Sulfonium-Ion Glucosidase Inhibitors and Their Derivatives: A Promising Class of Potential Antidiabetic Agents

In humans, four different enzymes mediate the digestion of ingested carbohydrates. First salivary and pancreatic α-amylases, the two endoacting retaining glucosidases, break down the complex starch molecules into smaller linear maltose-oligomers (LM) and branched α-limit dextrins (αLDx). Then two retaining exoglucosidases, maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI), convert those molecules into glucose in the small intestine. The small intestinal brush-border epithelial cells anchor MGAM and SI, and each contains a catalytic N- and C-terminal subunit, ntMGAM, ctMGAM, ntSI, and ctSI, respectively. All four catalytic domains have, to varying extents, α-1,4-exohydrolytic glucosidase activity and belong to the glycoside hydrolase family 31 (GH31). ntSI and ctSI show additional activity toward α-1,6 (isomaltose substrates) and α-1,2 (sucrose) glycosidic linkages, respectively. Because they mediate the final steps of starch digestion, both MGAM and SI are important target enzymes for the treatment of type-2 diabetes. Because of their potent inhibitory activities against the mammalian intestinal α-glucosidases, sulfonium-ion glucosidase inhibitors isolated from the antidiabetic herbal extracts of various Salacia species have received considerable attention recently. Thus far, researchers have isolated eight sulfonium-ion glucosidase inhibitors from Salacia species: salaprinol, salacinol, ponkoranol, kotalanol, and four of their corresponding de-O-sulfonated compounds, the structures of which comprise a 1,4-anhydro-4-thio-d-arabinitol and a polyhydroxylated acyclic side chain. Some of these compounds more strongly inhibit human intestinal α-glucosidases than the currently available antidiabetic drugs, acarbose and miglitol, and could serve as lead candidates in the treatment of type-2 diabetes. In this Account, we summarize progress in the field since 2010 with this class of inhibitors, with particular focus on their selective inhibitory activities against the intestinal glucosidases. Through structure-activity relationship (SAR) studies, we have modified the natural compounds to derive more potent, nanomolar inhibitors of human MGAM and SI. This structural optimization also yielded the most potent inhibitors known to date for each subunit. Furthermore, we observed that some of our synthetic inhibitors selectively blocked the activity of some mucosal α-glucosidases. Those results led to our current working hypothesis that selective inhibitors can dampen the action of a fast digesting subunit or subunits which places the burden of digestion on slower digesting subunits. That strategy can control the rate of starch digestion and glucose release to the body. Decreasing the initial glucose spike after a carbohydrate-rich meal and extending postprandial blood glucose delivery to the body can be desirable for diabetics and patients with other metabolic syndrome-associated diseases.