Isomaltooligosaccharides Research Articles

Prebiotic potential of novel carbohydrates in an in vitro co-inoculation fermentation model of the bacteria isolated from pig intestine and Salmonella

Innovative plant biomass fractionation methods produce new feed additives that could modulate pig intestinal microbial communities. Such novel indigestible carbohydrates (CHO) were investigated for their prebiotic potential and their influence on Salmonella Typhimurium in a co-inoculation in vitro fermentation model of pig intestines. Inulin, cellobiose, pectic oligosaccharides (POS), isomalto-oligosaccharides (IMO), xylooligosaccharides (XOS), and gluconic acid (GLU) were fermented for 72 h by a bacterial fecal inoculum from pigs in an in vitro model in the presence of Salmonella. Gluconic acid was the fastest fermenting CHO followed by inulin and IMO (P = 0.01). After 6 h, cellobiose yielded the highest lactate molar ratio (0.484). Pectic oligosaccharides fermented more slowly. Xylooligosaccharides and GLU were little fermented (150 and 175 mg short-chain fatty acids/g after 24 h). Nonetheless, GLU yielded the highest butyrate molar ratio of all CHO (0.290 at 12 h; P < 0.01). Although Salmonella counts did not differ, inulin and IMO displayed prebiotical properties, because they supported the highest Lactobacillus and Bifidobacterium populations after 12 and 24 h of fermentation (7.38 to 8.86 log cfu/mL; P < 0.01). Cellobiose and GLU scored well for Lactobacillus too but poorly supported Bifidobacterium (6.41 to 6.92 log cfu/mL; P < 0.01). It is concluded that IMO seems the most promising prebiotic, but owing to their specific fermentation patterns, cellobiose and GLU deserve further consideration.

Effect of fructo-oligosaccharide and isomalto-oligosaccharide addition on baking quality of frozen dough

The baking quality of frozen doughs containing different levels of fructo-oligosaccharides (FO) or isomalto-oligosaccharides (IMO) (3–9%, w/w flour), and stored for 0–8weeks at −18°C, was examined. The addition of FO or IMO increased the proof volume of the dough and the loaf volume of bread prepared from frozen dough. A 6% addition of FO or IMO was optimum, giving the highest proof volume and bread loaf volume, but a higher concentration than 6% induced low baking quality including lower proof volume and bread loaf volume. The bread crumb was moister and softer after the addition of FO or IMO before, and even after, frozen storage. Darker crumb colour was observed in the bread after the addition of FO or IMO. The oligosaccharides added to the frozen dough were effective in improving the quality of bread made from frozen dough, except for resulting in a darker bread crumb.

Production of isomaltooligosaccharides (IMO) using simultaneous saccharification and transglucosylation from starch and sustainable sources

Isomaltooligosaccharide (IMO), a well-known prebiotic, was produced from starch by a novel approach of simultaneous saccharification and transglucosylation (SST). The dosage of enzyme mixture consisting of Fungamyl 800L, for saccharification and α-glucosidase, for transglucosylation was optimized by Nelder-Mead simplex algorithm. The optimal conditions observed at the end of five iterations were 0.06U for Fungamyl 800L and 1.05U for α-glucosidase per 5mL of liquefied slurry. The production of IMO from soluble starch using these optimal concentration of enzymes in SST was found to be 95.08±1.22g/L at the end of 12h in comparison to 89.66±2.64 that was obtained using consecutive saccharification and translucosylation (CST) at the end of 24h. Thus produced IMO via SST comprised of (in g/L) isomaltose: 13.83, maltotriose: 5.58, panose: 39.40, tetrasaccharides (DP4): 16.1, pentasaccharides (DP5): 11.29 and other higher oligosaccharides (≥DP6): 8.86. The optimized SST process was also extrapolated to two sustainable starch sources viz., potato processing waste (PPW) and broken rice (BR). The IMO produced using PPW and BR as substrates were found to be 92.17±3.43g/L and 85.11±4.30g/L, respectively. These results indicate that SST could be a promising process intensification alternative for the sustainable production of IMO in comparison to existing methods.

Determination of phosphoryl‐oligosaccharides obtained from Canna edulis Ker starch

Phosphoryl oligosaccharides (POs) are a new type of functional oligosaccharide that promote the absorption of calcium. Canna edulis Ker (edible canna) starch contains a small amount of phosphorus and may be a source in the production of POs. Herein, the POs fraction was separated from Canna edulis Ker starch for the first time using enzymatic reactions that produce isomalto‐oligosaccharides (IMOs). Fourier transform infrared (FT‐IR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, and high‐performance anion exchange chromatography (HPAEC) were used to determine the presence and composition of the isolated POs. The POs that were obtained from Canna edulis Ker starch were mainly composed of phosphoryl maltotriose, maltotetraose, maltopentaose, maltohexaose, and maltoheptaose. These findings provide new insight into the simultaneous production of IMOs and POs from Canna edulis Ker starch by enzymatic reaction.

Optimization of Isomaltooligosaccharide Size Distribution by Acceptor Reaction of Weissella confusa Dextransucrase and Characterization of Novel α-(1→2)-Branched Isomaltooligosaccharides.

Long-chain isomaltooligosaccharides (IMOs) are promising prebiotics. IMOs were produced by a Weissella confusa dextransucrase via maltose acceptor reaction. The inputs of substrates (i.e., sucrose and maltose, 0.15-1 M) and dextransucrase (1-10 U/g sucrose) were used to control IMO yield and profile. According to response surface modeling, 1 M sucrose and 0.5 M maltose were optimal for the synthesis of longer IMOs, whereas the dextransucrase dosage showed no significant effect. In addition to the principal linear IMOs, a homologous series of minor IMOs were also produced from maltose. As identified by MS(n) and NMR spectroscopy, the minor trisaccharide contained an α-(1→2)-linked glucosyl residue on the reducing residue of maltose and thus was α-d-glucopyranosyl-(1→2)-[α-d-glucopyranosyl-(1→4)]-d-glucopyranose (centose). The higher members of the series were probably formed by the attachment of a single unit branch to linear IMOs. This is the first report of such α-(1→2)-branched IMOs produced from maltose by a dextransucrase.

In vitroselection of a synbiotic andin vivoevaluation on intestinal microbiota, performance and physiological response of rainbow trout (Oncorhynchus mykiss) fingerlings

The aim of this study was to determine the best synbiotic combination (based on growth and short chain fatty acids production) between Pediococcus acidilactici and five prebiotics in vitro, inulin, fructooligosaccharide (FOS), xylooligosaccharide (XOS), galactooligosaccharide (GOS), isomaltooligosaccharide (IMO). The best in vitro growth under aerobic and anaerobic culture condition was observed in the treatment, P. acidilactici + GOS. Furthermore, the study of short chain fatty acids (SCFA) production at the end of exponential growth phase revealed that the major SCFA produced was propionic acid in all synbiotic treatments except for the P. acidilactici + GOS treatment where butyric acid production was significantly higher compared with the other treatments. Based on the results of in vitro studies, P. acidilactici + GOS that were selected as synbiotic and rainbow trout fingerlings (15.04 ± 0.52 g) were fed on diet supplemented with the selected synbiotic. After 8 weeks of feeding, growth performance, intestinal microbiota and physiological responses were evaluated. Growth performance was significantly improved by synbiotic feeding. No significant differences of haematological parameters were revealed between the different treatments. Level of presumptive autochthonous lactic acid bacteria was elevated, while total autochthonous bacterial level remained unaffected. These results showed that P. acidilactici + GOS can be considered as an efficient synbiotic for rainbow trout.

Prebiotic preferences of human lactobacilli strains in co-culture with bifidobacteria and antimicrobial activity against Clostridium difficile.

To evaluate robustness, prebiotic utilization of Lactobacillus paracasei F8 and Lactobacillus plantarum F44 in mono- and co-cultures with Bifidobacterium breve 46 and Bifidobacterium animalis sub sp. lactis 8:8 and antimicrobial activity of co-culture against Clostridium difficile. The two Lactobacillus strains showed a high acid and bile tolerance. Lactobacillus plantarum F44 showed maximum growth in de Man Rogosa Sharpe basal broth with glucose and lactulose compared to growth in galacto-oligosaccharides (GOS) and isomalto-oligosaccharides (IMOS). In co-culture system, the amylolytic Bif.breve 46 stimulated the growth of a nonamylolytic Lact.paracasei F8, probably by producing intermediate metabolites of starch metabolism. A higher growth of four strains Lact.paracasei F8, Lact.plantarum F44, Bif.breve 46 and Bif.animalis ssp lactis 8:8 with different prebiotic combinations was found in a MRSC basal broth with SS (soluble starch) + IMOS + GOS and IMOS + GOS respectively. The two Lactobacillus strains exhibited a high antimicrobial activity against four clinical Cl.difficile strains and a hypervirulent NAP1/027strain and suppressed the toxin titres possibly through the production of organic acids and heat stable antimicrobial proteins when grown on glucose and through the production of acids when grown on prebiotics. Culture supernatants from synbiotic combinations inhibited the growth of the Cl.difficile NAP1/027 strain and its toxin titres. Lactobacillus paracasei F8, Lact.plantarum F44 exhibited potential probiotic properties. Further, the two Lactobacillus and two bifidobacteria strains were compatible with each other and exhibited high growth in co-cultures in presence of prebiotics and SS and antimicrobial activity against clinical Cl.difficile strains and a hypervirulent NAPI/027 strain. Results are promising for the development of a multi-strain synergistic synbiotic supplement for protection against Cl.difficile infection.

Enzyme-resistant isomalto-oligosaccharides produced from Leuconostoc mesenteroides NRRL B-1426 dextran hydrolysis for functional food application.

The extracellular dextransucrase from Leuconostoc mesenteroides NRRL B-1426 was produced and purified using polyethylene glycol fractionation. In our earlier study, it was reported that L. mesenteroides dextransucrase synthesizes a high-molecular mass dextran (>2×10(6) Da) with ∼85.5% α-(1→6) linear and ∼14.5% α-(1→3) branched linkages. Isomalto-oligosaccharides (IMOs) were synthesized through depolymerization of dextran by the action of dextranase. The degree of polymerization of IMOs was 2-10 as confirmed by mass spectrometry. The nuclear magnetic resonance spectroscopic analysis revealed the presence of α-(1→3) linkages in the synthesized IMOs. The IMOs were resistant to dextranase, α-glucosidase, and α-amylase, and therefore can have potential application as food additives in the functional foods.

Effects of Graded Levels of Isomaltooligosaccharides on the Performance, Immune Function and Intestinal Status of Weaned Pigs.

The objective of this study was to investigate the effects of graded levels of isomaltooligosaccharides (IMO) on the performance, immune function and intestinal microflora and intestinal mucosal morphology of weaned pigs. In a 28-day experiment, one hundred eighty, twenty eight-day-old, crossbred (Duroc×Large White×Landrace), weaned pigs, with an initial body weight of 8.19±1.45 kg, were fed either an unsupplemented corn-soybean meal based diet or similar diets supplemented with 0.2%, 0.4%, 0.6%, or 0.8% IMO added at the expense of corn. Each treatment was replicated six times with six pigs (three barrows and three gilts) per pen. From day 0 to 14, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p = 0.05) linearly declined as the IMO level increased. On d 14, the level of the immunoglobulins IgA, IgM, and IgG in the serum of pigs were linearly increased (p<0.05) with increasing IMO supplementation. Interleukin-6 (IL-6) was linearly (p<0.05) and quadratically (p<0.05) decreased as IMO intake increased. From day 15 to 28, there was a trend for weight gain to be linearly increased, and IL-2 was linearly (p<0.05) increased as IMO supplementation increased on d 28. Over the entire experiment, weight gain was linearly increased (p<0.05), while gain:feed (p<0.05) was linearly improved and diarrhea rate (p<0.05) was linearly decreased as the IMO level increased. Supplementation with IMO had no effect on the intestinal microflora of pigs in the ileum and cecum of pigs, as well as the villus height and crypt depth in the ileum and jejunum (p>0.05). These results indicate that dietary inclusion of IMO increases weight gain, gain:feed and enhanced the immune status of pigs, and could be a valuable feed additive for use in weaned pigs, particularly during the period immediately after weaning.

Preparation, Characterization, and Antioxidant Activity of an Isomaltooligosaccharide–Iron Complex (IIC)

GRAPHICAL ABSTRACT

Simple synthesis of isomaltooligosaccharides during Sauerkraut fermentation by addition of Leuconostoc starter and sugars

Isomaltooligosaccharides (IMOs), prebiotic compounds stimulating the growth of intestinal bacteria, were synthesized in sauerkraut by inoculating psychrotrophic Leuconostoc citreum KACC 91035 strain with high dextransucrase activity. For the glucose transferring reaction of dextransucrase, sucrose and maltose were added in sauerkraut (29 and 28 mM, respectively, w/v) as glucosyl donor and acceptor molecule, respectively. For 12 days of fermentation at 10℃, IMOs were gradually produced by consuming sucrose and maltose, and the synthesis rate and maximal concentration of IMOs were 2.04 and 20.2 mM, respectively. This result demonstrates a simple method to manufacture a synbiotic sauerkraut product by adding probiotic lactic acid bacterium and sugars as ingredients.

Synthesis of long-chain isomaltooligosaccharides from tapioca starch and an in vitro investigation of their prebiotic properties

The recombinant wild-type (WT) or Y101S-mutated amylomaltase prepared from the gene screened from soil DNA was used in combination with Aspergillus niger transglucosidase to produce isomaltooligosaccharides (IMOs) from tapioca starch. The highest IMO yield was obtained when 30% (w/v) soluble tapioca starch was incubated with 120 Units of both amylomaltases and 6 Units of transglucosidase at 40°C for 30min with WT or for 1h with Y101S-mutated enzymes. Mass spectrometry and 1H NMR analysis of the IMOs synthesized from WT and Y101S-mutated enzymes showed the presence of α-1,4 and α-1,6 glycosidic bonds with a polymerization degree of ≤9. HPAEC-PAD analysis showed that the sizes of the IMOs synthesized from both enzymes were significantly larger than those of commercial IMOs. A smaller proportion of long-chain IMOs was observed when Y101S-amylomaltase was used. As for physical and biological properties, the degree of sweetness of both synthesized IMOs was lower than those of sucrose and commercial IMOs, while the brown color from the Maillard reaction, the viscosity and the hygroscopicity were higher than those of commercial IMOs. The prebiotic properties of both synthesized IMOs showed that they were able to tolerate acidic conditions, heat, and human digestive enzymes. In addition, these IMOs could stimulate the growth of Lactobacillus casei and resulted in a decrease of the culture pH, similar to the effect of commercial IMOs.

Chemical Compositions of Egg Yolks and Egg Quality of Laying Hens Fed Prebiotic, Probiotic, and Synbiotic Diets.

A 16-wk feeding experiment was conducted to investigate the effects of a prebiotic, isomaltooligosaccharide (IMO), a probiotic, PrimaLac®, and their combination as a synbiotic on the chemical compositions of egg yolks and the egg quality of laying hens. One hundred and sixty 16-wk-old Hisex Brown pullets were randomly assigned to 4 dietary treatments: (i) basal diet (control), (ii) basal diet + 1% IMO (PRE), (iii) basal diet + 0.1% PrimaLac® (PRO), and (iv) basal diet + 1% IMO + 0.1% PrimaLac® (SYN). PRE, PRO, or SYN supplementation not only significantly (P < 0.05) decreased the egg yolk cholesterol (24- and 28-wk-old) and total saturated fatty acids (SFA; 28-, 32-, and 36-wk-old), but also significantly (P < 0.05) increased total unsaturated fatty acids (UFA; 28-, 32-, and 36-wk-old), total omega 6 and polyunsaturated fatty acids (PUFA), including linoleic and alpha-linolenic acid levels in the eggs (28-wk-old). However, the total lipids, carotenoids, and tocopherols in the egg yolks were similar among all dietary treatments in the 24-, 28-, 32-, and 36-wk-old hens. Egg quality (Haugh unit, relative weights of the albumen and yolk, specific gravity, shell thickness, and yolk color) was not affected by PRE, PRO, or SYN supplementation. The results indicate that supplementations with IMO and PrimaLac® alone or in combination as a synbiotic might be useful for improving the cholesterol content and modifying the fatty acid compositions of egg yolk without affecting the quality of eggs from laying hens between 24 and 36 wk of age.

Isomaltooligosaccharide increases the Lactobacillus rhamnosus viable count in Cheddar cheese

Five batches of Cheddar cheese were manufactured containing different levels of isomaltooligosaccharide (IMO) and a probiotic strain of Lactobacillus rhamnosus to study the effect of IMO on the survival of starter lactococci and probiotic micro‐organisms, on proteolytic patterns, cheese composition and sensory properties. The cheese was exposed to conditions simulating those found in the gastrointestinal tract to evaluate the survival of Lb. rhamnosus. Results demonstrated that the addition of Lb. rhamnosus and IMO did not affect the main compositional variables of Cheddar cheese. The counts of starter culture and probiotic organisms increased in cheese which contained Isomaltooligosaccharide (Batches 3, 4 and 5) more than in the control (Batches 1 and 2) during the fermentation. The probiotic counts in fresh cheese (B‐4) was 9.23 log10 cfu/g which was more than one log cycle greater than in the control (B‐2). The probiotic counts remained above 8 log10 cfu/g at the end of the manufacturing process. Primary proteolysis was not affected by the addition of probiotic bacteria and IMO, but the level of secondary proteolysis was slightly higher compared with the control group. The addition of IMO improved the texture and sensory quality of the cheese and the probiotic bacterium had the same effect. Under conditions that simulated the gastrointestinal tract, the probiotic bacteria in cheese (B‐4) exhibited good survival and remained above the recommended 6–7 log10 cfu/g.

Modeling of enzymatic production of isomaltooligosaccharides: a mechanistic approach

In the current investigation the production of isomaltooligosaccharides (IMO) by transglucosylation of maltose using α-glucosidase was modeled by a mechanistic approach. Parameters are estimated by a genetic algorithm followed by sensitivity analysis and experimental validation.

Quality Characteristics of Tangor Jam including Fructo Oligosaccharide and Isomalto Oligosaccharide

Quality Characteristics of Tangor Jam including Fructo Oligosaccharide and Isomalto Oligosaccharide

Quality Characteristics of Tangor Jam including Fructo Oligosaccharide and Isomalto Oligosaccharide

Quality Characteristics of Tangor Jam including Fructo Oligosaccharide and Isomalto Oligosaccharide

Pickering Emulsion Stabilized by Immobilized Bienzyme Nanoparticles: A Novel and Robust System for Enzymatic Purification of Isomaltooligosaccharide

A bienzyme system aiming at removal of glucose from isomaltooligosaccharide (IMO) was developed in this study. In this bienzymatic system, the first enzyme (glucose oxidase, GOD) catalyzed glucose to generate H2O2 and gluconic acid, while the second one (catalase, CAT) broke down the undesired H2O2 to water and oxygen, which can reduce the peroxide-induced degradation of GOD. For improving the catalytic activity, realizing the recyclability, and simplifying the separation operations of the enzymes, a Pickering emulsion stabilized by GOD- and CAT-containing silica particles was constructed. The optimal reaction temperature and pH were identified as 45 °C and 6.0, respectively. Under such conditions, the removal efficiency of glucose could be maintained at ca. 73.57%, even after five cycles in repeated batch operations. It is believed that the proposed process of glucose removal with biocatalytic Pickering emulsion is promising for the purification of IMO on an industrial scale.

Chemically defined diet alters the protective properties of fructo-oligosaccharides and isomalto-oligosaccharides in HLA-B27 transgenic rats.

Non-digestible oligosaccharides (NDO) were shown to reduce inflammation in experimental colitis, but it remains unclear whether microbiota changes mediate their colitis-modulating effects. This study assessed intestinal microbiota and intestinal inflammation after feeding chemically defined AIN-76A or rat chow diets, with or without supplementation with 8 g/kg body weight of fructo-oligosaccharides (FOS) or isomalto-oligosaccharides (IMO). The study used HLA-B27 transgenic rats, a validated model of inflammatory bowel disease (IBD), in a factorial design with 6 treatment groups. Intestinal inflammation and intestinal microbiota were analysed after 12 weeks of treatment. FOS and IMO reduced colitis in animals fed rat chow, but exhibited no anti-inflammatory effect when added to AIN-76A diets. Both NDO induced specific but divergent microbiota changes. Bifidobacteria and Enterobacteriaceae were stimulated by FOS, whereas copy numbers of Clostridium cluster IV were decreased. In addition, higher concentrations of total short-chain fatty acids (SCFA) were observed in cecal contents of rats on rat chow compared to the chemically defined diet. AIN-76A increased the relative proportions of propionate, iso-butyrate, valerate and iso-valerate irrespective of the oligosaccharide treatment. The SCFA composition, particularly the relative concentration of iso-butyrate, valerate and iso-valerate, was associated (P≤0.004 and r≥0.4) with increased colitis and IL-1 β concentration of the cecal mucosa. This study demonstrated that the protective effects of fibres on colitis development depend on the diet. Although diets modified specific cecal microbiota, our study indicates that these changes were not associated with colitis reduction. Intestinal inflammation was positively correlated to protein fermentation and negatively correlated with carbohydrate fermentation in the large intestine.

Mechanism-based antidiabetic activity of Fructo- and isomalto-oligosaccharides: Validation by in vivo, in silico and in vitro interaction potential

This study evaluates the relative beneficial effects of 10% dietary intake of fructooligosaccharides (FOSs) and isomaltooligosaccharides (IMOs) and combination of FOS+IMO in poloxamer-407 (PX-407) induced type 2 diabetic Wistar rats. FOSs was produced from Aspergillus oryzae (MTCC5154) while IMOs and standards of 1-kestose, 1-nystose, 1-fructofuranosyl nystose and panose were procured. In silico docking studies were performed by GLIDE program for each of the FOSs and IMOs for PPAR-γ activation and DPP-IV inhibition. Diabetic rats treated with FOS+IMO showed relatively more amelioration of glycemic and lipid dysmetabolism, remarkable reduction in oxidative markers, increased GLP-1 content as well as Bifidobacteria/Lactobacilli population in caecum than lone FOSs/IMOs treatment. Out of nine oligosaccharides docked from FOS and IMO; panose, nystose and kestose showed highest ranking binding mode with DPP-IV and PPAR-γ and were selected for in vitro study either alone or in combinations. On its own nystose showed potent DPP-IV inhibitory activity with an IC50 of 146.8μM while panose at 20.2μM concentrations showed 50% binding ability to PPAR-γ-LBD. Combinations of oligosaccharides tested namely Nys+Pan, Nys+Kes and Pan+Kes demonstrated significant (p<0.001) effect on PPAR-γ/DPP-IV bioassay. The results provide pharmacological evidence of FOSs and IMOs as antihyperglycemic mediated by their interaction with multiple targets operating in diabetes particularly nystose and pannose.