Raffinose/Sucrose/D-Glucose Assay Kit

Procedures for the measurement of starch, starch damage (gelatinised starch), resistant starch and the amylose/amylopectin content of starch, β-glucan, fructan, glucomannan and galactosyl-sucrose oligosaccharides (raffinose, stachyose and verbascose) in plant material, animal feeds and foods are described. Most of these methods have been successfully subjected to interlaboratory evaluation. All methods are based on the use of enzymes either purified by conventional chromatography or produced using molecular biology techniques. Such methods allow specific, accurate and reliable quantification of a particular component. Problems in calculating the actual weight of galactosyl-sucrose oligosaccharides in test samples are discussed in detail.

An American Association of Cereal Chemists/AOAC collaborative study was conducted to evaluate the accuracy and reliability of an enzyme assay kit procedure for measurement of total starch in a range of cereal grains and products. The flour sample is incubated at 95 degrees C with thermostable alpha-amylase to catalyze the hydrolysis of starch to maltodextrins, the pH of the slurry is adjusted, and the slurry is treated with a highly purified amyloglucosidase to quantitatively hydrolyze the dextrins to glucose. Glucose is measured with glucose oxidase-peroxidase reagent. Thirty-two collaborators were sent 16 homogeneous test samples as 8 blind duplicates. These samples included chicken feed pellets, white bread, green peas, high-amylose maize starch, white wheat flour, wheat starch, oat bran, and spaghetti. All samples were analyzed by the standard procedure as detailed above; 4 samples (high-amylose maize starch and wheat starch) were also analyzed by a method that requires the samples to be cooked first in dimethyl sulfoxide (DMSO). Relative standard deviations for repeatability (RSD(r)) ranged from 2.1 to 3.9%, and relative standard deviations for reproducibility (RSD(R)) ranged from 2.9 to 5.7%. The RSD(R) value for high amylose maize starch analyzed by the standard (non-DMSO) procedure was 5.7%; the value was reduced to 2.9% when the DMSO procedure was used, and the determined starch values increased from 86.9 to 97.2%.

Functional gastrointestinal disorders like diarrhea continue to affect children under five years in low-income countries. Incorporating health-enhancing bioactive compounds such as prebiotics in diet offers a promising solution. This study investigated prebiotic potential of oligosaccharides extracted from improved varieties of millet (Seremi 2, Naromil 2), sesame (Sesim 2, Sesim 3), soybean (Maksoy 3N, Maksoy 6N), and sorghum (Narosorg 2, Narosorg 4), commonly consumed in Uganda. These were compared to their respective indigenous variety. This study employed standardized methods for optical density measurement, culture preparation, and oligosaccharide extraction to evaluate prebiotic properties. We investigated whether plant-based oligosaccharides could enhance the effectiveness of probiotics, specifically Lactiplantibacillus plantarum (ATCC 14917) and Lacticaseibacillus rhamnosus (ATCC 7469), in antagonizing common enteric pathogens (Salmonella enterica subsp. enterica (ATCC 13076) and Shigella flexneri (ATCC 12022)). Approximately 4-8 log CFU/ml of each probiotic was incubated in 2% w/v oligosaccharide extracts at 37°C to evaluate the influence of the extracts on their growth, short-chain fatty acid (SCFA) production and antagonistic activity. Maximum cell density, which exceeded the minimum recommended probiotic cell density (6 log CFU/ml), was achieved during 24-h incubation period. The probiotics exhibited optimal growth in extracts of Sesim 2, Maksoy 3N, Narosorg 2 and indigenous millet variety resulting in a 68-84% increase in cell densities. The concentration of SCFA concentration was significantly higher (p < 0.05) in soybean-based oligosaccharides. Both probiotics antagonized growth of Salmonella and Shigella by more than 40% when cultured on Sesim 2, Maksoy 3N, Narosorg 2 and indigenous millet variety, while maintaining the probiotic cell densities above the minimum recommended level. These varieties show great potential as functional ingredients for developing synbiotic-rich foods to promote gut and public health. However, to evaluate the oligosaccharides prebiotic efficacy, in vitro fermentation using fecal microbiota and in vivo studies are necessary to determine gut microbiota changes and interactions.

High potential is attributed to the concomitant use of probiotics and prebiotics in a single food product, called “synbiotics”, where the prebiotic component distinctly favours the growth and activity of probiotic microbes. This study implemented a detailed comparison between the prebiotic effect of Fructooligosaccharides (FOSs) and Raffinose family oligosaccharides (RFOs) on the viable count of bacteria, hydrolysis into monosaccharides, the biosynthesis of short-chain fatty acids and sensory attributes of soymilk fermented with 1% (v/v) co-cultures of Lacticaseibacillus rhamnosus JCM1136 and Weissella confusa 30082b. The highest viable count of 1.21 × 10⁹ CFU/mL was observed in soymilk with 3% RFOs added as a prebiotic source compared with MRS broth with 3% RFOs (3.21 × 10⁸) and 3% FOS (6.2 × 10⁷ CFU/mL) when replaced against glucose in MRS broth. Raffinose and stachyose were extensively metabolised (4.75 and 1.28-fold decrease, respectively) in 3% RFOs supplemented with soymilk, and there was an increase in glucose, galactose, fructose (2.36, 1.55, 2.76-fold, respectively) in soymilk supplemented with 3% FOS. Synbiotic soymilk with 3% RFOs showed a 99-fold increase in methyl propionate, while the one supplemented with 3% FOS showed an increase in methyl butyrate. The highest acceptability based on the sensory attributes was for soymilk fermented with 2% RFOs + 2% FOS + 2% table sugar + 1% vanillin (7.87 ± 0.52) with high mouth feel, product consistency, taste, and flavour. This study shows that the simultaneous administration of soy with probiotic bacteria and prebiotic oligosaccharides like FOSs and RFOs enhance the synergistic interaction between them, which upgraded the nutritional and sensory quality of synbiotic soymilk.

This study investigated the effect of processing (roasting and malting) and crop variety on fermentable oligo-di-monosaccharides and polyols (FODMAPs) profile of millet, sorghum, soybean, and sesame varieties commonly consumed in Uganda. Two elite varieties and one indigenous variety for each crop were analyzed. Monosaccharide and polyols content was determined by HPLC-UV method, while disaccharides and oligosaccharide were determined using Megazyme kits. The elite varieties of soybean (Maksoy 3 N), Millet (Seremi 2) and sorghum (Narosorg 2) exhibited significantly (p < 0.05) lower oligosaccharide content compared to indigenous varieties with percentage differences ranging from 10.2 to 73.9%. Additionally, Maksoy 3 N and Narosorg 2 also exhibited significantly lower (p < 0.05) excess fructose content compared to the indigenous variety. Malting was more effective than roasting (p < 0.05) in reducing FODMAP categories and total FODMAP content. Malting effectively reduced excess fructose in all grain types to the recommended levels of <0.15 g/100 g compared to roasting. Moreover, malting reduced total oligosaccharides and total FODMAPs in soybean and sesame by more than 50%. However, this reduction did not achieve the recommended threshold of 0.3 g/100 g for total oligosaccharides and 0.5 g/100 g, for total FODMAPs which are a criterion to categorize low FODMAP diets. Malting conditions should be optimized to enhance its effectiveness in producing low FODMAP foods. This study highlights the importance of selecting appropriate grain variety and processing techniques that modify FODMAP content in foods that can be used for dietary therapy of gastro-intestinal disorders among vulnerable population.

Bioconversion processes represent sustainable, environmentally friendly, and cost-effective tools to improve the nutritional quality, bioactivity, and technological properties of agri-food waste. The chickpea cooking wastewater aquafaba, commonly used as an egg-replacer ingredient in various food formulations, was investigated as a suitable substrate for Lacticaseibacillus paracasei NPB-01's growth, which reached a final bacterial load of 9 Log and lactic acid production of 2.16 g/L after 24 h of process. Despite total saponins and polyphenols showing nonsignificant differences before and after fermentation, a significant improvement in the antioxidant power of fermented aquafaba was found. The microbial proteolysis and the simultaneous approach of pH to the chickpea proteins' isoelectric value (approximately 4.5) conferred high surface hydrophobicity and flexibility to the protein units, emphasizing the technological characteristics of aquafaba. In particular, a tenfold-enhanced emulsifying capacity and a significant improvement in foam and emulsion stability (98% and 100%, respectively) were observed, confirming the potential of fermented aquafaba as an enhanced texture-modifying ingredient with probiotic and antioxidant properties.

Among the species used for syrup production, sugar maple (Acer saccharum Marsh.) is preferred by producers, while red maple (Acer rubrum L.) is considered less productive in terms of sap yield and sugar content. This study aims to measure the volume and physicochemical characteristics of the sap produced from two red maples and two sugar maples during the 2023 sugar season in a commercial sugarbush in Laterrière (QC, Canada). Sap exudation was measured continuously with the gravity method using automatic rain gauges. Sap production was discontinuous and heterogeneous, reaching 2.6 L during the most productive day. No significant difference was found in the daily production between species, but we observed a difference in the cumulative sap production (7 L in red maple vs. 13.5 L in sugar maple) due to a longer period of sap exudation in the latter. Despite daily variations in pH, Brix values, sucrose concentration, osmolality, and conductivity, no differences in physicochemical characteristics were detected between species.

Grain chalkiness is an undesirable trait that negatively regulates grain yield and quality in rice. However, the regulatory mechanism underlying chalkiness is complex and remains unclear. We identified a positive regulator of white-belly rate (WBR). The WBR7 gene encodes sucrose synthase 3 (SUS3). A weak functional allele of WBR7 is beneficial in increasing grain yield and quality. During the domestication of indica rice, a functional G/A variation in the coding region of WBR7 resulted in an E541K amino acid substitution in the GT-4 glycosyltransferase domain, leading to a significant decrease in decomposition activity of WBR7^A (allele in cultivar Jin23B) compared with WBR7^G (allele in cultivar Beilu130). The NIL(J23B) and knockout line NIL(BL130)^KO exhibited lower WBR7 decomposition activity than that of NIL(BL130) and NIL(J23B)^COM, resulting in less sucrose decomposition and metabolism in the conducting organs. This caused more sucrose transportation to the endosperm, enhancing the synthesis of storage components in the endosperm and leading to decreased WBR. More sucrose was also transported to the anthers, providing sufficient substrate and energy supply for pollen maturation and germination, ultimately leading to an increase rate of seed setting and increased grain yield. Our findings elucidate a mechanism for enhancing rice yield and quality by modulating sucrose metabolism and allocation, and provides a valuable allele for improved rice quality.

In this work, the malting process of lentil seeds (Lens culinaris) was set-up to minimize their anti-nutrient content. The first (water steeping) and second (germination) process steps were studied in a 1.2-kg bench-top plant at 25°C. After 2-h steeping about 98.8% of seeds sprouted. As the germination process was prolonged for 72 h, the flatulence-inducing raffinose or phytic acid content was reduced by 94% or 63%, respectively. The third process step (kilning), carried out under fluent dry air at 50°C for 48 h and at 75°C for 3 h, gave rise to a gold metallic yellow-lentil malt, the cotyledons of which were cyclonically recovered and finally milled. The resulting decorticated yellow-lentil malt flour was used to prepare a fresh egg pasta high in raw protein (28±2 g/100 g), low in phytate (0.46±0.03 g/100 g) and in vitro glycemic index (38%), and approximately zero oligosaccharides.

The consumption of chickpeas is disadvantaged by their long cooking times and thus great cooking energy needs, as well as undesirable tastes and smells and the presence of some anti-nutritional factors. This work was aimed at measuring the cooking and nutritional characteristics of a typical chickpea variety (the straight furrow chickpea, SDC) cultivated in the Latium Region of Italy, as such, decorticated or after malting. A three-stage (steeping, germination and kilning) process allowed their original contents of α-galactosides and phytate to be reduced by about 57% and 31%, respectively. Once decorticated, malted SDCs were directly cooked in boiling water for about 45 min, while the 16-h presoaked raw counterpart needed a 30-min longer cooking process. Dehulled malted SDCs can thus assist the general consumer with shorter preparation times and more healthy and sustainable eating habits.

High potential is attributed to the concomitant use of probiotics and prebiotics in a single food product, called "synbiotics," where the prebiotic component distinctly favours the growth and activity of probiotic microbes. This study implemented a detailed comparison between the prebiotic effect of Fructooligosaccharides (FOS) and Raffinose Oligosaccharides (RFOs) on the viable count of bacteria, hydrolysis into monosaccharides for the biosynthesis of postbiotic Short Chain Fatty Acids and sensory attributes of soymilk fermented with 1% (v/v) mixed cultures of Lacticaseibacillus rhamnosus JCM 1136 and Weissella confusa 30082b. The highest viable count of 1.21×10⁹ CFU/ml was observed in soymilk with 3%RFOs added as a prebiotic source compared with MRS broth with 3% RFOs (3.21×10⁸) and 3%FOS (6.2 ×10⁷ CFU/ml) when replaced against glucose in MRS broth. Raffinose and stachyose were extensively metabolised (4.75 and 1.28-fold decrease, respectively) in 3%RFOs supplemented with soymilk, and there was an increase in glucose, galactose, fructose (2.36,1.55, 2.76-fold respectively) in soymilk supplemented with 3% FOS. Synbiotic soymilk with 3%RFOs showed a 99-fold increase in methyl propionate, while the one supplemented with 3%FOS showed an increase in methyl butyrate. The highest acceptability was for soymilk fermented with 2%RFOs + 2%FOS + 2% Table sugar + 1% vanillin (7.87 ± 0.52) with high Mouth feel, product consistency, taste, and flavour. This study shows that simultaneous administration of soy with probiotic bacteria and prebiotic oligosaccharides like FOS and RFOs enhance the synergistic interaction between them, which up-graded the nutritional and sensory quality of synbiotic soymilk.

Our study investigated the effectiveness of 446 strains of lactic acid bacteria (LAB) belonging to different species and isolated from diverse sources (food, human, and animal) as potential probiotic candidates, with the perspective of producing dietary supplements or pharmacological formulations suitable for enhancing gastrointestinal digestion. The survival capability of all the isolates under harsh gastrointestinal tract conditions was evaluated, in which only 44 strains, named high-resistant, were selected for further food digestibility investigations. All 44 strains hydrolyzed raffinose and exhibited amino and iminopeptidase activities but at various extents, confirming species- and strain-specificity. After partial in vitro digestion mimicking oral and gastric digestive phases, food matrices were incubated with single strains for 24 h. Fermented partially digested matrices provided additional functional properties for some investigated strains by releasing peptides and increasing the release of highly bio-accessible free phenolic compounds. A scoring procedure was proposed as an effective tool to reduce data complexity and quantitively characterize the probiotic potential of each LAB strain, which could be more useful in the selection procedure of powerful probiotics.

The composition and properties of potatoes have been comprehensively studied; however, effects of different cultivation periods on these factors remain unclear. Hence, we aim to provide insights for cultivating potatoes with desirable properties by controlling the potato growth time and cultivars. Physico-chemical quality characteristics of three potato cultivars-Chubaek, Superior, and Atlantic-were analysed at different cultivation periods (80, 90, and 100 days). Regardless of cultivation period, total starch and amylose content of the cultivars was as follows: Atlantic > Superior > Chubaek, whereas sucrose and glucose content was in reverse order. The Atlantic cultivar showed the highest dry matter content, whereas Chubaek showed the highest moisture content. In all cultivars, glutamic acid and aspartic acid were the predominant amino acids. Peak viscosity was the highest in Atlantic cultivar and increased as the cultivation period progressed. Final, peak, and breakdown viscosities were positively correlated with starch and amylose content and negatively correlated with glucose, sucrose, and most amino acid content. Hierarchical clustering heatmaps showed that all three cultivars cultivated for 100 days exhibited different patterns from those cultivated for 80 and 90 days. Overall, quality characteristics of the potatoes were more influenced by genetic characteristics than by the cultivation period.

Raffinose family oligosaccharides (RFOs) are known to have important physiological functions in plants. However, the presence of RFOs in legumes causes flatulence, hence are considered antinutrients. To reduce the RFOs content to a desirable limit without compromising normal plant development and functioning, the identification of important regulatory genes associated with the biosynthetic pathway is a prerequisite. In the present study, through comparative RNA sequencing in contrasting genotypes for seed RFOs content at different seed maturity stages, differentially expressed genes (DEGs) associated with the pathway were identified. The DEGs exhibited spatio-temporal expression patterns with high RFOs variety showing early induction of RFOs biosynthetic genes and low RFOs variety showing a late expression at seed maturity. Selective and seed-specific differential expression of raffinose synthase genes (AhRS14 and AhRS6) suggested their regulatory role in RFOs accumulation in peanut seeds, thereby serving as promising targets in low RFOs peanut breeding programs. Despite stachyose being the major seed RFOs fraction, differential expression of raffinose synthase genes indicated the complex metabolic regulation of this pathway. The transcriptomic resource and the genes identified in this study could be studied further to develop low RFOs varieties, thus improving the overall nutritional quality of peanuts.

Compositional analyses of sequentially pearled fractions of high-tannin (HT, Athena) and low-tannin (LT, Snowbird) faba bean (Vicia faba L.) cultivars demonstrated that the decreasing trend of protein, ash and total dietary fibre (TDF) contents from outer to inner layers was offset by starch contents. LT and HT proteins were positively correlated (r = 0.9, P < 0.0001) with ash and total dietary fibre contents while negatively correlated (r = −0.9, P < 0.0001) with starch contents with increasing degree of pearling. The 55-57 % single-step pearling flour had a higher (P < 0.05) protein content (LT:39.07 ± 0.06 % and HT:37.37 ± 0.03 %, N × 6.25) as compared to whole beans (LT:30.51 ± 0.39 and HT:29.63 ± 0.20 %). The starch contents of the pearled beans were significantly higher (P < 0.05) than whole beans. These single-step pearling flour fractions subjected to aqueous fractionation to isolate proteins (>94 % purity) resulted in starch isolates and dietary fibre concentrates as co-products. Hybrid fractionation had minimal impact on the native protein secondary structures where β-sheets were dominant. This study suggests that the inclusion of pearling as an upstream processing step prior to wet fractionation of both LT and HT faba beans has the potential to generate novel “clean label” ingredients for the food industry.