Analysis of bioaccessibility of campesterol,stigmasterol, and β-sitosterol in maize by in vitro digestion method

The content of campesterol, stigmasterol, and β-sitosterol in raw and cooked Big Flint Maize (BFM), Pop Corn Maize (PCM) and Red Maize (RM) were analyzed by using a chemical method and using in vitro digestion of cooked samples. RM contained 15.14, 8.36, and 72.19 mg/100 g campesterol, stigmasterol, and β-sitosterol, respectively, which were higher compared to in BFM and PCM. Samples cooking resulted in 6–48% increase in phytosterol content, while cooked RM had the highest phytosterol content. Using the in vitro digestion method, 44–61% reduction in phytosterol content was observed in contrast to the chemical method. The difference bioaccessible campesterol and β-sitosterol contents among varieties was insignificant, except for stigmasterol in RM. The bioaccessibility of campesterol was 44–56% higher than those of stigmasterol and β-sitosterol, and it was higher in BFM and PCM. In conclusion, phytosterol contents estimated in different maize samples using in vitro digestion can be another determinative for the ultimate content and reveals the necessity to consider with chemical method of determination.     

Impact of native form oat β-glucan on starch digestion and postprandial glycemia

Whole grain oats, as a rich source of β-glucan, have been shown beneficial to glycemic control. In the current study, the impact of native form β-glucan in oat grains (NFO-glucan) on starch digestion and postprandial glycemia was investigated. The dry-milling prepared NFO-glucan sample was enriched with native form β-glucan (15.6%), and an in vitro starch digestion assay of NFO-glucan (0.5% starch equivalent) showed a significant decrease of starch digestion rate compared to oat starch (0.5%, w/v). However, pretreatment by either β-glucanase or pepsin significantly increased the starch digestion. Consistently, an in vivo examination on the postprandial glycemia of the cooked NFO-glucan sample using a mouse model displayed a significant decrease of postprandial glycemia compared to gelatinized oat starch. Further experiment on the pasting property of NFO-glucan sample by a rapid visco-analyser demonstrated both β-glucan and protein affected its viscosity profiles. Scanning Electronic Microscope (SEM) observation revealed a network-like native structure of β-glucan that might encapsulate protein and starch to reduce the enzyme accessibility and so the digestion of starch. Novel food processing technologies to maintain the native form of β-glucan in oat grains might be a better way to modulate the postprandial glycemia of oat-based whole grain foods.     

Pre-maturity α-amylase in wheat: The role of abscisic acid and gibberellins

The occurrence of pre-maturity α-amylase (PMA) is a major cause of poor bread-making quality (low Hagberg Falling Number) in wheat grain. In susceptible genotypes, it involves the excessive accumulation of high isoelectric point (pI) α-amylase in mature grain prior to germination and in the absence of pre-harvest sprouting. Several factors regulate PMA formation in developing grain, including genotype, agronomy, and environmental conditions. In particular, a cold period during mid-grain development has been found to be a major stimulus for PMA induction. Although the factors affecting the PMA occurrence are well known, little is known about the molecular mechanism governing its induction. The plant hormones abscisic acid (ABA) and gibberellins (GAs) influence various aspects of grain development, and it has been suggested that PMA involves changes in the amount of these hormones or the sensitivity of the grain to these hormones. This review summarizes recent studies investigating the role of ABA and GAs in PMA induction and PMA occurrence.     

Significant down-regulation of γ-gliadins has minor effect on gluten and starch properties of bread wheat

The contribution of gliadins to the baking performance of wheat has been widely discussed. In the present study we report the properties of the flours of fifteen transgenic wheat lines with γ-gliadins down-regulated by RNAi. Technological properties were assessed by sodium dodecyl sulfate sedimentation (SDSS) test, and rheological properties were characterized during mixing using the Mixolab^®. Changes in the proportions of gluten proteins, with significant increases of glutenins, were observed in transgenic lines. These changes did not affect the SDSS test in most of the transgenic lines. The dough strength of some transgenic lines was reduced as determined by the Mixolab^®. Changes in the starch behavior were also observed in the transgenic lines, in which lower values of torque were observed at the C3, C4 and C5 points of the mixing curve. The results reported here are important in understanding the role of γ-gliadins in the bread-making quality of flours.     

The role of sensitivity to abscisic acid and gibberellin in pre-maturity α-amylase formation in wheat grains

To study the role of abscisic acid (ABA) and gibberellin (GA) sensitivity in regulating pre-maturity α-amylase (PMA) in wheat grains, plants were grown in a glasshouse under cold-shock and ambient conditions. α-amylase activity in response to applied ABA and GA was measured in detached-grains with the embryo removed (in vitro) and in intact-grains attached to the plant (in situ). The in vitro experiment was conducted using Spark (low PMA-susceptible genotype) and Rialto (highly PMA-susceptible genotype), with the aim of defining the time point for GA-sensitivity. The results showed an increase in GA-sensitivity at about 640 degree days after anthesis (DAA) in Rialto. There was no evidence for a change in ABA-sensitivity in either variety. The in situ experiments were conducted using genotypes from a Spark × Rialto doubled haploid population segregating for the Rht-D1a (tall) or Rht-D1b allele and for the presence or absence of 1BS/1RS. For Rht-D1a (tall) or Rht-D1b genotypes with or without 1BS/1RS, the cold-shock significantly increased GA-sensitivity, whereas there was no significant change in ABA-sensitivity. These results show PMA is related to an increase in GA-sensitivity that occurs in the aleurone at around 640 degree DAA, and can be enhanced by environmental factors (e.g. cold-shock).     

The role of sensitivity to abscisic acid and gibberellin in pre-maturity α-amylase formation in wheat grains

To study the role of abscisic acid (ABA) and gibberellin (GA) sensitivity in regulating pre-maturity α-amylase (PMA) in wheat grains, plants were grown in a glasshouse under cold-shock and ambient conditions. α-amylase activity in response to applied ABA and GA was measured in detached-grains with the embryo removed (in vitro) and in intact-grains attached to the plant (in situ). The in vitro experiment was conducted using Spark (low PMA-susceptible genotype) and Rialto (highly PMA-susceptible genotype), with the aim of defining the time point for GA-sensitivity. The results showed an increase in GA-sensitivity at about 640 degree days after anthesis (DAA) in Rialto. There was no evidence for a change in ABA-sensitivity in either variety. The in situ experiments were conducted using genotypes from a Spark × Rialto doubled haploid population segregating for the Rht-D1a (tall) or Rht-D1b allele and for the presence or absence of 1BS/1RS. For Rht-D1a (tall) or Rht-D1b genotypes with or without 1BS/1RS, the cold-shock significantly increased GA-sensitivity, whereas there was no significant change in ABA-sensitivity. These results show PMA is related to an increase in GA-sensitivity that occurs in the aleurone at around 640 degree DAA, and can be enhanced by environmental factors (e.g. cold-shock).     

Biological and chemical evaluation of chick pea seed proteins as affected by germination,extraction and α-amylase treatment

The effects of germination, extraction (double extraction with 70% ethanol and water at isoelectric point) and -amylase treatments of chick pea seed flours on crude protein, total carbohydrate, protein efficiency ratio (PER), biological value (BV), true digestibility (TD), net protein utilization (NPU), essential amino acid composition, in-vitro protein digestibility (IVPD) and actual amino acid indices (essential amino acid index or amino acid score) were evaluated. Crude protein content was increased (8–149%), while total carbohydrate was decreased (11–62%) by germination, extraction and -amylase treatments. Alpha-amylase treatment was more efficient in reducing total carbohydrate and increasing the protein content than that of extraction treatment. The protein quality of chick pea flours as measured by PER, BV, TD, NPU, IVPD and corrected amino acid indices (actual amino acid indices×IVPD) was significantly improved by these treatments. The protein quality of germinated--amylase treatment was comparble with casein, while germinated--amylase treaded seeds appeared nutritionally superior to casein. The results indicate that the germinated--amylase and germinated--amylase-extracted treatments could be used successfully as a source of concentrated high quality protein for baby food production. The corrected amino acid indices gave better prediction of PER, BV, TD and NPU (r=93 to 97) than actual amino acid indices (r=45 to 71). PER was highly correlated with corrected amino acid score (r=0.93). The PER could be predicted from the following simple regression equation: PER=–1.827+0.0561×corrected amino acid score.     

Inhibition study of red rice polyphenols on pancreatic α-amylase activity by kinetic analysis and molecular docking

This study sought to investigate the possible inhibition mechanism of red rice polyphenols (RRP) on pancreatic α-amylase (PA) activity. RRP showed strong inhibition against PA activity and the half-inhibitory concentration (IC₅₀) value was 3.61 μg/mL. The fluorescence quenching of PA by RRP was a combination of static quenching and dynamic quenching. RRP could aggregate with PA and the physiochemical properties of the aggregates were closely related to the concentration of RRP. Kinetic analysis suggested that the inhibition mode of RRP on PA was reversible inhibition, which was a mixing of competitive inhibition and noncompetitive inhibition. Molecular docking speculated that RRP could form hydrogen bonds with PA by binding to the catalytic active sites (ASP197, GLU233 and ASP300) and the microenvironments of TRP58 and TRP59 were altered, thus inhibiting PA activity.     

Inhibition of α-amylase purified fromOctopus vulgaris Lam. by albumin inhibitors from wheat flour

Octopus -amylase has been purified to homogeneity by single-step affinity chromatography on Sepharose-bound wheat albumin inhibitors. Two electrophoretically distinguishable isoamylases are obtained, both consisting of a single polypeptide chain with molecular weight 45,000. Octopus -amylase is more effectively inhibited by a monomeric than by a dimeric protein inhibitor from wheat. The inhibition is dependent on pH, temperature, and time of preincubation. Properties are compared with those of other animal -amylases.     

Two β-amylase genes,OsBAM2 and OsBAM3, are involved in starch remobilization in rice leaf sheaths

To identify mechanisms of starch degradation in rice leaf sheaths at the post-heading stage, we investigated the function of OsBAM2 and OsBAM3, which encode plastid-targeted active β-amylase isoforms, in starch remobilization in leaf sheaths. The starch content in the second leaf sheaths below the flag leaf (the third leaf sheaths) peaked at the flag leaf emergence stage and gradually decreased until 15 days after heading. The mRNA levels of OsBAM2 and OsBAM3 in the third leaf sheaths increased from the flag leaf emergence stage to the heading stage when the starch content began to decrease. However, these mRNA levels did not always remain high during post-heading. Overexpression of OsBAM2 or OsBAM3 markedly repressed starch accumulation in the third leaf sheaths, showing that OsBAM2 and OsBAM3 function in starch degradation in rice leaf sheaths. In contrast, no significant differences in starch content in the third leaf sheaths were detected between knockdown plants of OsBAM2 or OsBAM3 and non-transgenic wild-type plants. Our results suggest that reduced expression of the individual genes, OsBAM2 or OsBAM3, does not result in excess accumulation of starch in the leaf sheaths, probably because of the complementary function of another gene or the action of other genes encoding starch-degrading .     

The heterogeneous distribution of α-amylase and endoxylanase activity over a population of preharvest sprouted wheat kernels and their localization in individual kernels

To develop targeted approaches to improve the quality of preharvest sprouted (PHS) wheat as a raw material for food manufacturing, knowledge on the nature and distribution of hydrolytic enzymes in PHS wheat is crucial. Results of the present study indicate that α-amylase and endoxylanase activities are heterogeneously distributed among a population of PHS kernel. Within individual severely sprouted kernels, the enzyme activities are heterogeneously distributed throughout the different tissues. α-Amylase activity, almost exclusively of endogenous nature, is mainly detected in the germ region and to a lesser extent in the aleurone layer. Endoxylanase activity is predominantly of microbial origin and located on the kernel surface. In spite of this, light and epifluorescence microscopy show decreased kernel integrity and cell wall breakdown in the crushed cells layer, the endosperm, and the aleurone layer in a selection of kernels upon preharvest sprouting. This knowledge offers opportunities for the development of treatments to reduce the enzyme load in PHS wheat at postharvest level to improve its flour quality.     

Inhibitory Activity of Cinnamon Bark Species and their Combination Effect with Acarbose against Intestinal α-glucosidase and Pancreatic α-amylase

Inhibition of α-glucosidase and pancreatic α-amylase is one of the therapeutic approaches for delaying carbohydrate digestion, resulting in reduced postprandial glucose. The aim of this study was to evaluate the phytochemical analysis and the inhibitory effect of various cinnamon bark species against intestinal α-glucosidase and pancreatic α-amylase. The results showed that the content of total phenolic, flavonoid, and condensed tannin ranged from 0.17 to 0.21 g gallic acid equivalent/g extract, from 48.85 to 65.52 mg quercetin equivalent/g extract, and from 0.12 to 0.15 g catechin equivalent/g extract, respectively. The HPLC fingerprints of each cinnamon species were established. Among cinnamon species, Thai cinnamon extract was the most potent inhibitor against the intestinal maltase with the IC₅₀ values of 0.58 ± 0.01 mg/ml. The findings also showed that Ceylon cinnamon was the most effective intestinal sucrase and pancreatic α-amylase inhibitor with the IC₅₀ values of 0.42 ± 0.02 and 1.23 ± 0.02 mg/ml, respectively. In addition, cinnamon extracts produced additive inhibition against intestinal α-glucosidase and pancreatic α-amylase when combined with acarbose. These results suggest that cinnamon bark extracts may be potentially useful for the control of postprandial glucose in diabetic patients through inhibition of intestinal α-glucosidase and pancreatic α-amylase.     

Associations between caryopsis dormancy, α-amylase activity,and pre-harvest sprouting in barley

Pre-harvest sprouting (PHS) is a concern for barley (Hordeum vulgare L.) producers, grain processors, and researchers worldwide. Pre-harvest sprouting has been mainly attributed to low dormancy, which is determined by genotype, stage of plant maturation, and environmental conditions during caryopsis development. Fourteen barley genotypes were sown in field experiments at two sites in North Dakota in 2004 and 2005. Spikes were harvested at four different stages: ≈500 g kg⁻¹ moisture content, physiological maturity, harvest maturity, and post-harvest maturity. Results indicated that barley genotypes were released from dormancy at different rates. The 14 barley genotypes were divided into three classes based on their dormancy loss rate during caryopsis development. C93-3230-24 was highly dormant, and ‘Stander’ and ‘Legacy’ were highly susceptible to PHS due to lack of dormancy from as early as 20 d after heading date. All other genotypes fell into the third group that had intermediate dormancy loss rate. No significant correlation was detected between barley α-amylase activity and germination percentage. A moderate association between malt α-amylase activity and caryopsis dormancy suggested that cultivars with increased malt α-amylase activity tend to have low dormancy and may be more prone to PHS.     

Polymorphism and pedigree analysis of β-amylase isozymes in Australian wheat

β-Amylase encoded by multiple loci on chromosomes of triticeae crop plants is a starch metabolic enzyme with a significant physiological role in flour functionality. In this study, wheat grain β-amylase isozymes were characterized using an extensive Australian wheat population and genetic breeding/mapping lines. β-amylase isozymes were fractionated by high-resolution native polyacrylamide gel electrophoresis (PAGE) and validated by zymogram and peptide sequencing using electrospray ionization tandem mass spectrometry (ESI-MS/MS). Specific antibodies were generated against the β-amylase purified through protein chromatography and used for β-amylase identification. Further characterization of wheat β-amylases was carried out with immunoblotting and 2-dimensional electrophoresis (2-DE). The enzyme was found to be polymorphic among the Australian and foreign wheat cultivars, with a total of six isozymes: two common and four varying isozymes, the latter group including one novel β-amylase isozyme of fast-mobility in native PAGE, According to pedigree analysis, the origin of this particular isozyme was from the ancestral genotype, Ciano F 67.     

Studies on α-amylase and trypsin inhibitors in legume seeds using agar diffusion and isoelectric focusing techniques

Amylolytic and tryptic inhibitors of faba bean extracts were determined by an agar diffusion test. The amylolytic inhibitor had protein characters. Furthermore, water-soluble trypsin inhibitors ofCicer arietinum, Lens esculenta, Lupinus termis, Phaseolus vulgaris, Pisum sativum, Trigonella foenum-graecum andVicia faba which were separated by polyacrylamide gel isoelectric focusing (PAGIF) in thin-layers, showed species specific patterns. Negative staining showed 10 bands for French beans, 9 for fenugreek seeds, 8 for lentils and chickpeas, 7 for peas and 6 for faba beans. Lupin seeds were free from trypsin inhibitors. Treatments (soaking, germination and heat processing) of faba beans reduced the number of trypsin inhibitors in PAGIF patterns, less after soaking and germination, but more after roasting and frying. No inhibitors were detected after cooking.     

Investigating the impact of α-amylase, α-glucosidase and glucoamylase action on yeast-mediated bread dough fermentation and bread sugar levels

Wheat flour is generally supplemented with α-amylases to increase maltose levels in bread dough and increase loaf volume. While the preference of yeast for glucose and fructose over maltose as substrate for fermentation is well documented, the impact of maltose versus glucose producing enzymes on bread dough fermentation kinetics and bread sugar levels is ill documented. Hence the impact of α-amylase, α-glucosidase and glucoamylase action on both aspects was investigated. Glucoamylase and α-amylase increase the total fermentable sugar content of dough, while α-glucosidase only affects the glucose/maltose ratio. Due to their effect on total fermentable sugar levels, addition of α-amylase or glucoamylase prolongs the total productive fermentation time, while this is not the case for α-glucosidase. In contrast to α-amylase, both glucoamylase and α-glucosidase supplementation leads to higher CO₂ production rates during the initial stages of fermentation. In the final bread product, different sugar levels are observed depending on the dosage and type of starch-degrading enzyme. The results of this study imply that long and short fermentation processes benefit from α-amylase and α-glucosidase addition, respectively, while glucoamylase supplementation is suitable for both long and short fermentation times.     

Effects of selected extrusion parameters on physicochemical properties and in vitro starch digestibility and β-glucan extractability of whole grain oats

Whole grain oat flour was extruded under different moisture contents (15%, 18%, 21%), barrel temperatures (100 °C, 130 °C), and screw speeds (160 rpm, 300 rpm, 450 rpm), and selected physicochemical properties, in vitro starch digestibility, and β-glucan extractability of the extrudates were analyzed. An increase in screw speed resulted in an increase in radial expansion index, water absorption index, and water solubility index. Screw speed significantly affected slowly and rapidly digestible starch. Moderate screw speed (300 rpm) led to higher slowly digestible starch with an accompanying decrease in rapidly digestible starch. Low moisture conditions (15%) resulted in the highest resistant starch and water-extractable β-glucan. Under the conditions used in this study, extrusion did not result in changes in water-extractable β-glucan molecular weight. Thus, extrusion might be beneficial in improving functionality and consumer acceptability by affecting physicochemical properties, in vitro starch digestibility, and β-glucan extractability of oat extrudates.