Mechanisms Underlying the Effect of Tea Extracts on In Vitro Digestion of Wheat Starch.

The effect of extracts from four types of tea made from Camelia sinensis (green, white, black, and oolong) on in vitro amylolysis of gelatinized starch and the underlying mechanisms were studied. Of the four extracts, black tea extract (BTE) gave the strongest inhibition of starch digestion and on α-amylase activity. Fluorescence quenching and surface plasmon resonance (SPR) showed compounds in BTE bound to α-amylase more strongly than those in the green, white, and oolong tea extracts. Individual testing of five phenolic compounds abundant in tea extracts showed that theaflavins had a greater inhibitory effect than catechins on α-amylase. SPR showed that theaflavins had much lower equilibrium dissociation constants and therefore bound more tightly to α-amylase than catechins. We conclude that BTE had a stronger inhibitory effect on in vitro enzymatic starch digestion than the other tea extracts, mainly due to the higher content of theaflavins causing stronger inhibition of α-amylase.

Inhibition of in vitro enzymatic starch digestion by coffee extract.

There is evidence that moderate coffee consumption is beneficial in the prevention of type 2 diabetes, however, the underlying mechanism is not understood. In this study, the effects of an extract of ground coffee on the in vitro enzymatic digestion of starch were investigated. The coffee extract decreased the rate and extent of starch digestion, with kinetic analysis showing that the extract reduced the binding affinity of the enzymes for the substrate and their catalytic turnover. Fluorescence quenching indicated that the coffee extract formed complexes with the digestive enzymes through a static quenching mechanism. Ultraviolet absorption and circular dichroism spectra of the digestive enzymes confirmed that the coffee extract decreased the proportion of ß-sheet structures in the enzymes. Therefore, we conclude that compounds in the soluble coffee extract can interact with porcine pancreatic amylase and amyloglucosidase causing inhibition of the enzymes and decreasing in vitro starch digestion.

In vitro digestibility of starches with different crystalline polymorphs at low α-amylase activity to substrate ratio.

In this study, potato, lotus seed and wheat starch samples with different degree of gelatinization (DG) were prepared and their in vitro digestibility at low α-amylase activity evaluated by measuring the release of reducing sugar. The hydrolysis rate (k) and the final equilibrium concentration (C∞) of the three starches increased with increasing DG. Kinetic analyses showed that the Michaelis-Menten constant (Km) and the catalytic efficiency (kcat/Km) increased with increasing DG, indicative of the increasing affinity and catalytic efficiency of α-amylase with all three starch samples. Of the three starches, lotus seed starch showed a much greater increase in k and kcat/Km than potato and wheat starches as the DG of starch increased. From this study, we concluded that at low activity of α-amylase, DG is a major determinant for the binding affinity and catalytic efficiency of α-amylase to starch and in turn the digestion rate of starch.

Revealing the mechanisms of starch amylolysis affected by tea catechins using surface plasmon resonance.

The interaction of polyphenolic catechins from Camellia sinensis tea with α-amylase, and the effects of the interactions on the kinetics of starch amylolysis, were investigated. Binding studies using surface plasmon resonance (SPR) and enzyme kinetic analysis indicated that the in vitro digestibility of gelatinized wheat starch was decreased by the addition of catechins present in tea. Tea catechins decreased enzyme activity by reducing the maximum velocity (Vmax) of the α-amylase, but had little effect on the Michaelis-Menten constant (Km). Binding studies by SPR showed that the structure of the catechins influenced their affinity for α-amylase. Pearson correlation analysis showed that the decreased digestibility of starch in the presence of catechins was due to their binding of α-amylase interaction inhibiting the catalytic effectiveness of the enzyme. From this study, we concluded that the faster and stronger the binding of catechins and α-amylase, the greater reduction of starch digestion is.

Revisiting Mechanisms Underlying Digestion of Starches.

The factors that determine the digestion rate of starches were revealed using different forms of starches and a mixture of α-amylase and amyloglucosidase. Gelatinized starch samples with a degree of gelatinization (DG) from 12.2 to 100% for potato starch and from 7.1 to 100% for lotus seed starch were obtained. With an increasing DG, the short- and long-range molecular orders of both starches were disrupted progressively. The first-order digestion rate constant (k) of both starches increased with an increasing DG, although the positive linear relationships between DG and k differed (R2 = 0.87 for potato starch, and R2 = 0.74 for lotus seed starch). The mean fluorescence intensity showed a positive linear correlation with DG, which was strong for potato starch (R2 = 0.99) and relatively weaker for lotus seed starch (R2 = 0.54). These results indicated that DG is a major determinant for the digestion rate of potato starch and lotus seed starch and that the access/binding of enzymes to starch was the main rate-limiting factor for digestion of starches.

Effect of CaCl2 pre-treatment on the succinylation of potato starch.

Potato starch was pre-treated with CaCl2 solutions prior to modification with octenyl succinic anhydride (OSA). Starch pre-treated with 1.0 M CaCl2 showed higher degree of substitution (DS) and reaction efficiency (RE) on OSA modification, whereas pre-treatment with CaCl2 solutions at 0.05 M, 0.1 M and 0.5 M had no effect on DS and RE. CaCl2 pre-treatment decreased the swelling power, paste clarity, peak viscosity (PV), breakdown (BD) and some textural parameters of potato starch, with the effects being greater at higher concentrations of CaCl2. Pre-treatment with 1.0 M CaCl2 caused a small disruption to starch crystallinity and granule morphology. OSA modification significantly decreased the textural parameters, PV, BD, relative crystallinity, swelling power, gelatinization temperatures and enthalpy of potato starch, but it increased the paste clarity and emulsifying activity. OSA-1.0 M-starch showed improved functional properties over OSA-starch, indicating that CaCl2 pre-treatment provides advantages for improving the functional characters of succinylated starch.

Mechanisms Underlying the Formation of Complexes between Maize Starch and Lipids.

This study aimed to reveal the mechanism of formation of complexes between native maize starch (NMS) and different types of lipids, namely palmitic acid (PA), monopalmitate glycerol (MPG), dipalmitate glycerol (DPG), and tripalmitate glycerol (TPG). The complexing index followed the order of MPG (96.3%) > PA (41.8%) > TPG (8.3%) > DPG (1.1%), indicating that MPG formed more complexes with NMS than PA, and that few complexes were formed between NMS and DPG and TPG. The NMS-PA complex presented higher thermal transition temperatures and lower enthalpy change than the NMS-MPG complex, indicating that although MPG formed more starch complexes, they had less stable crystalline structures than the complex between NMS and PA. X-ray diffraction (XRD) and Raman spectroscopy showed that both MPG and PA formed V-type crystalline structures with NMS, and confirmed that no complexes were formed between NMS and DPG and TPG. We conclude that the monoglyceride formed more starch-lipid complex with maize starch than PA, but that the monoglyceride complex had a less stable structure than that formed with PA. The di- and triglycerides did not form complexes with maize starch.

Properties of starch from potatoes differing in glycemic index.

Potatoes are a popular source of dietary carbohydrate worldwide and are generally considered to be a high glycemic index (GI) food. Potato starch characteristics play a key role in determining their rate of digestion and resulting glycemic response. Starches isolated from seven potato cultivars with different GI values, including a low GI cultivar (Carisma), were examined for relative crystallinity, granule size distribution, amylopectin chain length, and thermal and pasting properties. Starch from the Carisma cultivar was more thermally stable and more resistant to gelatinization, with significantly higher (p < 0.05) pasting temperature and differential scanning calorimetry (DSC) gelatinization onset, peak and conclusion temperatures, compared to the other cultivars. Differences between the potatoes in the other properties measured did not align with the GI ranking. Thermal analysis and starch pasting properties may be useful indicators for preliminary identification of potato cultivars that are digested slowly and have a lower GI.

Discovery of a low-glycaemic index potato and relationship with starch digestion in vitro.

Potatoes are usually a high-glycaemic index (GI) food. Finding a low-GI potato and developing a screening method for finding low-GI cultivars are both health and agricultural priorities. The aims of the present study were to screen the commonly used and newly introduced cultivars of potatoes, in a bid to discover a low-GI potato, and to describe the relationship between in vitro starch digestibility of cooked potatoes and their in vivo glycaemic response. According to International Standard Organisation (ISO) guidelines, seven different potato cultivars were tested for their GI. In vitro enzymatic starch hydrolysis and chemical analyses, including amylose content analysis, were carried out for each potato cultivar, and correlations with the respective GI values were sought. The potato cultivars had a wide range of GI values (53-103). The Carisma cultivar was classified as low GI and the Nicola cultivar (GI = 69) as medium GI and the other five cultivars were classified as high GI according to ISO guidelines. The GI values were strongly and positively correlated with the percentage of in vitro enzymatic hydrolysis of starch in the cooked potatoes, particularly with the hydrolysis percentage at 120 min (r 0·91 and P <0·01). Amylose, dietary fibre and total starch content was not correlated with either in vitro starch digestibility or GI. The findings suggest that low-GI potato cultivars can be identified by screening using a high-throughput in vitro digestion procedure, while chemical composition, including amylose and fibre content, is not indicative.

Neanderthal medics? Evidence for food, cooking, and medicinal plants entrapped in dental calculus.

Neanderthals disappeared sometime between 30,000 and 24,000 years ago. Until recently, Neanderthals were understood to have been predominantly meat-eaters; however, a growing body of evidence suggests their diet also included plants. We present the results of a study, in which sequential thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) and pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) were combined with morphological analysis of plant microfossils, to identify material entrapped in dental calculus from five Neanderthal individuals from the north Spanish site of El Sidrón. Our results provide the first molecular evidence for inhalation of wood-fire smoke and bitumen or oil shale and ingestion of a range of cooked plant foods. We also offer the first evidence for the use of medicinal plants by a Neanderthal individual. The varied use of plants that we have identified suggests that the Neanderthal occupants of El Sidrón had a sophisticated knowledge of their natural surroundings which included the ability to select and use certain plants.