Oligosaccharides Raffinose Research Articles

Purification and characterization of α-galactosidases from Penicillium griseoroseum for efficient soymilk hydrolysis

Soybean utilization is limited by the presence of raffinose oligosaccharides (RFO), which are not digested by humans and cause gastrointestinal discomfort. This study explores the potential of α-galactosidases from Penicillium griseoroseum for RFO hydrolysis in soymilk. Two distinct α-galactosidase enzymes, designated α-Gal1 and α-Gal2, were purified using a combination of ion-exchange chromatography and native polyacrylamide gel electrophoresis. Both enzymes exhibited characteristics of multimeric proteins and displayed similar biochemical properties. Optimal activity was observed at a pH range of 4.5–5.0 and a temperature range of 40–45 °C. Notably, α-Gal1 demonstrated high thermostability with a half-life of 16 h at 40 °C. The α-galactosidases displayed different substrate affinitiesfor the substrates ρ-NP-αGal, o-NP-αGal, rD-raffinose, d-stachyose, and mD-melibiose. The Michaelis-Menten constant (Km) values for α-Gal1 were 1.06, 1.31, 28.74, 19.88, and 4.77 mmol/L, respectively, while those for α-Gal2 were 0.8, 1.26, 30.46, 21.74 and 5.01 mmol/L, respectively. Both α-Gal1 and α-Gal2 were strongly inhibited by metal ions (Ag⁺, Cu2⁺, Fe2⁺, and Hg2⁺) and moderately inhibited by d-melibiose. Importantly, both enzymes efficiently hydrolyzed RFOs, achieving complete d-stachyose elimination from soymilk after a 6-h incubation. These findings propose the promising application of these α-galactosidases in industrial soymilk production, potentially enhancing its nutritional value and alleviating gastrointestinal issues in consumers.

Effect of spray-drying or fermentation on the solubility and carbohydrate profile of chickpea hydrolysates for beverage formulation

Powder beverage bases were developed using extruded and hydrolyzed chickpea supernatant through freeze-drying. This study evaluated the effects of spray-drying and adjuvant incorporation for producing powders and supernatant fermentation with LAB strains prior freeze-drying on the carbohydrate composition, structural integrity, and physical properties of chickpea beverage base powders. All powders had low insoluble (<0.2%) and soluble (<3%) dietary fiber contents due to processing. Starch content decreased below 9.49% when using adjuvants and 10.61% during fermentation. Both treatments also reduce raffinose oligosaccharide content below 6.1 mM/100 g. Spray-drying and adjuvant addition promoted a more amorphous starch structure, enhancing solubility, while fermentation induced more crystallinity, according to FTIR data. Microscopy showed small particle clusters and lack of birefringence. SEM images revealed starch granule damage from both fermentation and spray-drying, with more uniform particles in spray-dried powders. Spray-drying resulted in the smallest particle sizes (455-457 nm) and higher absolute zeta potential (8.12-9.51), indicating greater stability. Fermentation had the opposite effect, negatively impacting the colloidal system stability. Samples had a pH close to 6, except fermented powders with a pH of 3.6, affecting colloidal stability. Fermentation significantly reduced solubility but without practical implications, as all samples had solubility values over 93%, which is desirable for beverage-based powders. Spray-drying and fermentation altered the carbohydrate profile and structure of the powders. Fermentation prior producing powders may provide benefits, but it slightly decreases the system stability. Spray-drying can be used instead of freeze-drying to produce stable chickpea base powders that are highly soluble.

Hydrolysis and diffusion of raffinose oligosaccharides family products in chickpeas, lentils, and beans under different pH and temperature steeping conditions

Soaking pulses in water is a traditional practice widely used both by many households and by the food industry, and depending on the specific conditions used, can effectively reduce α-galactosides. Monitoring changes in α-galactoside content in pulses under different steeping conditions can provide insights into the degradation mechanisms and help overcome the barrier to consumption caused by digestive problems. In this study, we analyzed the impact of steeping at different temperatures (30, 45, 60, 75, and 90 °C) and at different pH (4.0, 5.0, and 6.0) on α-galactosides content in chickpeas, lentils, and beans. Our results showed that the lower the pH, the faster the α-galactosides were reduced. Moreover, steeping at lower temperatures (30 °C and 45 °C) favored hydrolysis of α-galactosides, whereas steeping at higher temperatures (60, 75, and 90 °C) favored diffusion. Soaking at 45 °C at a pH of 4.0 for 3 h resulted in acceptable levels of α-galactosides (less than 1 g/100 g), i.e. a reduction of up to 65 % in chickpeas, 85 % in lentils, and 52 % in beans.

From Genes to Stress Response: Genomic and Transcriptomic Data Suggest the Significance of the Inositol and Raffinose Family Oligosaccharide Pathways in Stylosanthes scabra, Adaptation to the Caatinga Environment.

S. scabra is an important forage and extremophilic plant native to the Brazilian Caatinga semiarid region. It has only recently been subjected to omics-based investigations, and the generated datasets offer insights into biotechnologically significant candidates yet to be thoroughly examined. INSs (inositol and its derivatives) and RFO (raffinose oligosaccharide family) pathways emerge as pivotal candidates, given their critical roles in plant physiology. The mentioned compounds have also been linked to negative impacts on the absorption of nutrients in mammals, affecting overall nutritional intake and metabolism. Therefore, studying these metabolic pathways is important not just for plants but also for animals who depend on them as part of their diet. INS and RFO pathways in S. scabra stood out for their abundance of identified loci and enzymes. The enzymes exhibited genomic redundancy, being encoded by multiple loci and various gene families. The phylogenomic analysis unveiled an expansion of the PIP5K and GolS gene families relative to the immediate S. scabra ancestor. These enzymes are crucial for synthesizing key secondary messengers and the RFO precursor, respectively. Transcriptional control of the studied pathways was associated with DOF-type, C2H2, and BCP1 transcription factors. Identification of biological processes related to INS and RFO metabolic routes in S. scabra highlighted their significance in responding to stressful conditions prevalent in the Caatinga environment. Finally, RNA-Seq and qPCR data revealed the relevant influence of genes of the INS and RFO pathways in the S. scabra response to water deprivation. Our study deciphers the genetics and transcriptomics of the INS and RFO in S. scabra, shedding light on their importance for a Caatinga-native plant and paving the way for future biotechnological applications in this species and beyond.

An auxin regulated Universal stress protein (TaUSP_3B-1) interacts with TaGolS and provides tolerance under drought stress and ER stress.

A previously identified wheat drought stress responsive Universal stress protein, TaUSP_3B-1 has been found to work in an auxin dependent manner in the plant root tissues in the differentiation zone. We also found a novel interacting partner, TaGolS, which physically interacts with TaUSP_3B-1 and colocalizes in the endoplasmic reticulum. TaGolS is a key enzyme in the RFO (Raffinose oligosaccharides) biosynthesis which is well reported to provide tolerance under water deficit conditions. TaUSP_3B-1 overexpression lines showed an early flowering phenotype under drought stress which might be attributed to the increased levels of AtTPPB and AtTPS transcripts under drought stress. Moreover, at the cellular levels ER stress induced TaUSP_3B-1 transcription and provides tolerance in both adaptive and acute ER stress via less ROS accumulation in the overexpression lines. TaUSP_3B-1 overexpression plants had increased silique numbers and a denser root architecture as compared to the WT plants under drought stress.

Modulation of the nutritional and functional values of common bean by farming system: organic vs. conventional

Common bean (Phaseolus vulgaris L.) is a crop of high nutritional interest which is widespread throughout the world as a result of which it is considered as a potential crop for ensuring future food security. The effect of organic and conventional farming systems on the nutritional and functional composition of two cultivars of market class fabada (a landrace and a modern elite cultivar) is described. The findings of our study have shown that the common beans from organic farming have a higher content of protein, phenolic compounds, and relevant microelements such as iron and zinc than the seeds from conventional farming trial. However, in conventional crops a higher content of polyunsaturated fatty acids and lower concentrations of phytic acid and raffinose oligosaccharides were also detected. In contrast, no significant differences were detected in the proximate composition of the two genotypes tested, nor in their antioxidant activity or levels of phenolic compounds. The elite cultivar maintains the nutritional characteristics of the landrace from which it derives, suggesting the genomic regions modified in the improvement do not play an important role in their genetic control.

Far-Red Light Mediated Carbohydrate Concentration Changes in Leaves of Sweet Basil, a Stachyose Translocating Plant.

Photosynthetic active radiation (PAR) refers to photons between 400 and 700 nm. These photons drive photosynthesis, providing carbohydrates for plant metabolism and development. Far-red radiation (FR, 701-750 nm) is excluded in this definition because no FR is absorbed by the plant photosynthetic pigments. However, including FR in the light spectrum provides substantial benefits for biomass production and resource-use efficiency. We investigated the effects of continuous FR addition and end-of-day additional FR to a broad white light spectrum (BW) on carbohydrate concentrations in the top and bottom leaves of sweet basil (Ocimum basilicum L.), a species that produces the raffinose family oligosaccharides raffinose and stachyose and preferentially uses the latter as transport sugar. Glucose, fructose, sucrose, raffinose, and starch concentrations increased significantly in top and bottom leaves with the addition of FR light. The increased carbohydrate pools under FR light treatments are associated with more efficient stachyose production and potentially improved phloem loading through increased sucrose homeostasis in intermediary cells. The combination of a high biomass yield, increased resource-use efficiency, and increased carbohydrate concentration in leaves in response to the addition of FR light offers opportunities for commercial plant production in controlled growth environments.

The content of raffinose oligosaccharides in legumes and their importance for animals

The content of raffinose oligosaccharides in legumes and their importance for animals

Effects of Different Oligosaccharides on Growth Performance and Intestinal Function in Broilers.

ObjectiveThis study was conducted to investigate the effects of different oligosaccharides on the growth performance and intestinal function in broilers.MethodsA total of 360 1-day-old yellow-feather chickens were randomly divided into 5 groups and fed with a basal diet supplemented with 50 mg/kg chlortetracycline (ANT), 3 g/kg isomalto-oligosaccharide (IMO), 3 g/kg raffinose oligosaccharide (RFO), and 30 mg/kg chitooligosaccharide (COS). The experiment lasted for 56 days, with 1–28 days as the starter phase and 29–56 days as the grower phase.ResultsThe results showed that dietary supplementation with RFO and COS significantly improved average daily gain (ADG) and average daily feed intake (ADFI) (p < 0.05). Relative to the control group, diets supplemented with oligosaccharides dramatically increased the level of serum IgM (RFO, COS), T-SOD (COS), and GSH-Px (IMO and RFO) and the expression of ZO-1(IMO) and claudin-1 (RFO) (p < 0.05). Adding antibiotics or oligosaccharides to the diet could remarkedly increase the villus height and villus height (VH)/crypt depth (CD) ratio of each group (p < 0.05). Through the ileum α-diversity analysis and comparison of OTU number in each group showed that the microbial richness of the IMO group increased in the starter phase, and that of the RFO and CSO group increased in the grower phase. Additionally, compared with the control group, IMO supplementation increased the level of ileum sIgA (p < 0.05) and the content of valeric acid (p < 0.05) in the cecum.ConclusionsIn summary, the addition of oligosaccharides in diet can improve the immune function and antioxidant capacity and improve intestinal health of broilers.

Suppression of SDP1 Improves Soybean Seed Composition by Increasing Oil and Reducing Undigestible Oligosaccharides.

In developing soybean seeds, carbon is partitioned between oil, protein and carbohydrates. Here, we demonstrate that suppression of lipase-mediated turnover of triacylglycerols (TAG) during late seed development increases fatty acid content and decreases the presence of undigestible oligosaccharides. During late stages of embryo development, the fatty acid content of soybean seed decreases while the levels of the oligosaccharides raffinose and stachyose increase. Three soybean genes orthologous to the Arabidopsis lipase gene SUGAR-DEPENDENT1 (SDP1) are upregulated at this time. Suppression of these genes resulted in higher oil levels, with lipid levels in the best lines exceeding 24% of seed weight. In addition, lipase-suppressed lines produced larger seeds compared to wild-type plants, resulting in increases of over 20% in total lipid per seed. Levels of raffinose and stachyose were lower in the transgenic lines, with average reductions of 15% in total raffinose family oligosaccharides observed. Despite the increase in oil, protein content was not negatively impacted and trended higher in the transgenic lines. These results are consistent with a role for SDP1 in turning over TAG to supply carbon for other needs, including the synthesis of oligosaccharides, and offer new strategies to further improve the composition of soybean seeds.

Contrasting Metabolic Fingerprints and Seed Protein Profiles of Cucurbita foetidissima and C. radicans Fruits from Feral Plants Sampled in Central Mexico.

Cucurbita foetidissima and C. radicans are scarcely studied wild pumpkin species that grow in arid and semi-arid areas of Mexico and the United States. This study describes the morphological, proximal composition, metabolic finger-prints and seed protein profiles of C. foetidissima and C. radicans fruits collected in the wild during a one-year period in different locations of central-western Mexico. The results obtained complement the limited information concerning the fruit composition of C. foetidissima and greatly expand information in this respect regarding C. radicans. Morphology and proximal composition of their fruits varied significantly. Different metabolic fingerprints and seed protein profiles were detected between them and also with the chemical composition of domesticated Cucurbita fruits. The neutral lipids in seed, pulp and peels were rich in wax content and in unsaturated compounds, probably carotenoids and tocopherols, in addition to tri-, di- and mono-acylglycerols. The tri- and diacylglycerol profiles of their seed oils were different from commercial seed oils and between each other. They also showed unusual fatty acid compositions. Evidence of a possible alkaloid in the pulp and peel of both species was obtained in addition to several putative cucurbitacins. An abundance of phenolic acids was found in all fruit parts, whereas flavonoids were only detected in the peels. Unlike most cucurbits, globulins were not the main protein fraction in the seeds of C. radicans, whereas the non-structural carbohydrate and raffinose oligosaccharide content in their fruit parts was lower than in other wild cucurbit species. These results emphasize the significantly different chemical composition of these two marginally studied Cucurbita species, which was more discrepant in C. radicans, despite the notion regarding C. foetidissima as an aberrant species with no affinity to any other Cucurbita species.

Design and Characterization of a Novel Fermented Beverage from Lentil Grains.

The experimental activities carried out in this study aimed at designing a lentil-based beverage rich in soluble and digestible proteins. In order to extract soluble proteins, lentil grains were soaked in water overnight, blended, treated with proteolytic enzymes and fermented with Lactobacillus strains. Protein enzymatic hydrolysis, carried out with four commercial food grade enzyme preparations, showed that bromelin, at the enzyme to substrate ratio of 10%, was the best solution to produce this novel beverage. Even though the seven Lactobacillus strains were all able to ferment aqueous extract within 24 h, L. acidophilus ATCC 4356, L. fermentum DSM 20052 and L. paracasei subsp. paracasei DSM 20312 showed the highest growth rate and the lowest pH values. In fermented lentil-based beverages, the antinutritional factor phytic acid decreased up to 30%, similarly, the highest reduction in raffinose oligosaccharides content reached about 12% the initial concentration. It is worthy of note that the viable density of all strains remained higher than 7 log cfu/mL after 28 days of cold storage. The results here reported show for the first time the possibility to obtain a probiotic lentil-based beverage rich in soluble proteins, peptides and amino acids with low content in main antinutritional factors.

Integrated Proteomics and Metabolomics Analysis Highlights Correlative Metabolite-Protein Networks in Soybean Seeds Subjected to Warm-Water Soaking.

Soaking of soybean seeds is a prerequisite for the production of soy foods, and it has been shown that the extent of water absorbed during different imbibition conditions directly affects the quality of the subsequent soybean seed products by yet unknown mechanisms. In order to elucidate the molecular changes in soybean seeds during different soaking temperatures, we performed an integrated proteomics and metabolomics analysis of seeds soaked at 4, 25, and 55 °C. Proteomics analysis revealed that various enzymes related to carbohydrate and protein hydrolysis were activated in soybean seeds during water soaking at 55 °C. Interestingly, results obtained from this integrated proteomics and metabolomics study showed changes in various metabolites, including isoflavones, amino acids, and sugars, that were positively correlated with proteome changes occurring upon soaking at 55 °C. Furthermore, soaking of soybean seeds at 55 °C resulted in degradation of indigestible anti-nutrients such as raffinose oligosaccharides. Taken together, our results suggest that the seed soaking at a high temperature (55 °C) increases the nutritional value of soybean seeds by decreasing the contents of some of the common anti-nutrients.

Determining 1-kestose, nystose and raffinose oligosaccharides in grape juices and wines using HPLC: method validation and characterization of products from Northeast Brazil.

The objective of this work was to validate a method for direct determination in grape juice and wine of 1-kestose, nystose and raffinose oligosaccharides by reversed-phase high-performance liquid chromatography with refractive index detection using a new type of RP-C18 column (150 × 4.6mm, 4µm) with polar end-capping. The validated methodology was also used to characterize grape juice and fine wine products from Northeastern Brazil; and presented suitable linearity, precision, recovery, limits of detection and quantification. The method presented good specificity, revealing that sugars, organic acids, and ethanol (the main interferences in refraction detection) did not influence the quantification of the studied oligosaccharides. The main oligosaccharide found was 1-kestose (approximately 50% of the samples), followed by raffinose (20% of the samples). The results obtained in this are an indication that grape juices and wines have the potential to be functional beverages in relation to the presence of prebiotics.

Expression of &lt;i&gt;Oryza sativa&lt;/i&gt; galactinol synthase gene in maize (Zea may L.)

Galactinol synthase (GolS) is a key biological catalyst for the synthesis of the raffinose oligosaccharides (RFOs) which play important roles in abiotic stress adaptation of plants, especially drought tolerance. GolS gene has been isolated on a variety of plants in order to create material resources for generating transgenic plants resistant to adverse environmental factors. In our previous research, we have isolated a GolS gene from drought stress cDNA library of Oryza sativa L. Moctuyen (named OsGolS). In this study, the expression vector pCAM-Rd/OsGolS carrying the isolated OsGolS gene under the control of stress-inducible Rd29A promoter was constructed and introduced into Agrobacterium tumefaciens LBA4404, which was used for maize transformation. PCR and Real-time PCR assay indicated that transgene was integrated in the genome of the regenerated Zea mays plants. Reverse transcription-PCR showed that the OsGolS was transcribed into mRNA in Zea mays and was highly expressed. These results provide a basis for the study of the function of OsGolS in drought responses and for the development of drought stress tolerant crops.

Effect of Lactic Acid Bacteria Starter Cultures on Vitamin and Oligosaccharide Composition of Milk Extracted from Three Common Bean (Phaselous Vulgaris L) Varieties

Fermented foods have in recent times attracted consumer interest mainly due to perceived health benefits of probiotic microorganisms. This study characterized changes in the concentrations of selected B-complex vitamins and oligosaccharides of common bean milk during fermentation by a common dairy starter culture, YF L-903 (Streptococcus thermophilus + Lactobacillus Bulgaricus subs Debulgaricus), and three probiotic cultures namely ABT (Lactobacillus acidophilus La-5 + Bifidobacterium animalis Bb-12 + Streptococcus thermophilus), Yoba (Lactobacillus rhamnosus yoba + Streptococcus thermophilus), and Yoba Fiti (Lactobacillus rhamnosus GR1 + Streptococcus thermophilus). Bean milk was prepared from three common bean varieties. It was found that, apart from thiamine (vitamin B1) and riboflavin (vitamin B2), fermentation with each of the mixed cultures caused significant increase in the vitamin B complex. Significant reductions (p&amp;lt;0.05) in the oligosaccharides concentration of the bean milks were observed upon fermentation. Highest reduction in the oligosaccharide sugars of 77.8% was found in milk from pinto bean variety fermented with ABT culture. These findings suggest that LAB probiotic cultures have a potential for improving biosynthesis of vitamins and removal of the verbascose, stachyose and raffinose oligosaccharides, thus making the product more digestible and the nutrients more bioavailable.

Stability of the Metabolite Signature Resulting from the MIPS1 Mutation in Low Phytic Acid Soybean ( Glycine max L. Merr.) Mutants upon Cross-Breeding.

The low phytic acid ( lpa) soybean ( Glycine max L. Merr.) mutant Gm-lpa-TW-1-M, resulting from a 2 bp deletion in GmMIPS1, was crossed with a commercial cultivar. F3 and F5 progenies were subjected to nontargeted GC-based metabolite profiling, allowing analysis of a broad array of low molecular weight constituents. In the homozygous lpa mutant progenies the intended phytic acid reduction was accompanied by remarkable metabolic changes of nutritionally relevant constituents such as reduced contents of raffinose oligosaccharides and galactosyl cyclitols as well as increased concentrations in sucrose and various free amino acids. The mutation-induced metabolite signature was nearly unaffected by the cross-breeding and consistently expressed over generations and in different growing seasons. Therefore, not only the primary MIPS1 lpa mutant but also its progenies might be valuable genetic resources for commercial breeding programs to produce soybean seeds stably exhibiting improved phytate-related and nutritional properties.

Chromatographic fingerprinting of Lupinus luteus L. (Leguminosae) main secondary metabolites: a case of domestication affecting crop variability

Secondary metabolites (SMs), such as alkaloids and raffinose oligosaccharides (RFOs), play important roles in plant physiology. Although alkaloid and RFO phenotypic variation has been reported for yellow lupin (Lupinus luteus L.), most evaluations have used a reduced number of accessions; thus, limiting the understanding of accumulation patterns and variation ranges. The main goal of this research was to assess alkaloid and RFO content in a diverse L. luteus sample to understand possible SM accumulation patterns across this legume species. Eighteen yellow lupin accessions were analyzed using high performance thin layer chromatography to provide insights on seed and leaf RFO and alkaloid phenotypic variation. Co-dominant markers (170) were used to examine genetic relationships among L. luteus accessions and possible accumulation patterns across closely related genotypes. Significant differences were observed for seed and leaf RFOs. Total seed RFO accumulation ranged from 79.738 to 131.079 mg g−1. Raffinose, stachyose, and verbascose were observed in all genotypes’ seeds, but at different RFO concentrations. Raffinose was the only RFO detected in leaves (2.793–0.4224 mg g−1). Total alkaloid accumulation ranged from 0.22 to 15.12 and 0.00 to 8.007 mg g−1 for seeds and leaves, respectively. Lupinine, sparteine, and gramine were observed in seeds and leaves, and showed a wide range of variation. Neighbor-Joining (NJ) analysis showed an apparent pattern of seed alkaloid accumulation, most likely due to domestication events. However, high RFO accumulating accessions were scattered across the NJ tree. Alkaloid and RFO significant phenotypic variation will not only help to understand the roles of these SMs in L. luteus, but also to uncover the genetic basis behind their accumulation.

Exploring traditional aus-type rice for metabolites conferring drought tolerance

BackgroundTraditional varieties and landraces belonging to the aus-type group of rice (Oryza sativa L.) are known to be highly tolerant to environmental stresses, such as drought and heat, and are therefore recognized as a valuable genetic resource for crop improvement. Using two aus-type (Dular, N22) and two drought intolerant irrigated varieties (IR64, IR74) an untargeted metabolomics analysis was conducted to identify drought-responsive metabolites associated with tolerance.ResultsThe superior drought tolerance of Dular and N22 compared with the irrigated varieties was confirmed by phenotyping plants grown to maturity after imposing severe drought stress in a dry-down treatment. Dular and N22 did not show a significant reduction in grain yield compared to well-watered control plants, whereas the intolerant varieties showed a significant reduction in both, total spikelet number and grain yield. The metabolomics analysis was conducted with shoot and root samples of plants at the tillering stage at the end of the dry-down treatment. The data revealed an overall higher accumulation of N-rich metabolites (amino acids and nucleotide-related metabolites allantoin and uridine) in shoots of the tolerant varieties. In roots, the aus-type varieties were characterised by a higher reduction of metabolites representative of glycolysis and the TCA cycle, such as malate, glyceric acid and glyceric acid-3-phosphate. On the other hand, the oligosaccharide raffinose showed a higher fold increase in both, shoots and roots of the sensitive genotypes. The data further showed that, for certain drought-responsive metabolites, differences between the contrasting rice varieties were already evident under well-watered control conditions.ConclusionsThe drought tolerance-related metabolites identified in the aus-type varieties provide a valuable set of protective compounds and an entry point for assessing genetic diversity in the underlying pathways for developing drought tolerant rice and other crops.

Isolation of a protease-resistant and pH-stable α-galactosidase displaying hydrolytic efficacy toward raffinose family oligosaccharides from the button mushroom Agaricus bisporus

A 45-kDa monomeric acidic α-galactosidase with a specific activity of 193.12 units/mg was isolated from the fruiting bodies of Agaricus bisporus. Blast search of internal peptide sequences suggested that it is a member of GH family 27 and it is most similar to hypothetical protein AGABI2DRAFT_70106. The enzyme displayed maximal activity at pH 4.0 and 60°C, respectively. The enzyme remained stable within the pH range 2.0–9.0 but its activity was markedly suppressed in the presence of Cu2+, Hg2+, Fe3+ and Ag+ ions. It displayed resistance to α-chymotrypsin and neutral protease. Moreover, it manifested degradative activity toward both oligosaccharides and polysaccharides. The enzyme manifested Km values of 0.30mM, 10.65mM and 19.21mM, toward pNPGal, stachyose and raffinose respectively. These results suggest that Agaricus bisporus α-galactosidase is a promising candidate for elimination of raffinose oligosaccharides (RFOs) in biotechnological applications.