Lactobacillus acidophilus PIN7 paraprobiotic supplementation ameliorates DSS-induced colitis through anti-inflammatory and immune regulatory effects.

AIMS: This study aimed to evaluate the efficacy of paraprobiotics Lactobacillus acidophilus PIN7 supplementation against dextran sodium sulphate (DSS)-induced colitis in mice and to determine their mechanisms of the action. METHODS AND RESULTS: Ten-week-old female BALB/C mice were randomly divided into five groups. Each group was administered with PBS (control and DSS group), live PIN7 (LIVE group), heat-killed PIN7 (HEAT group) or lysozyme-treated PIN7 (LYSOZYME group) for 10 days followed by 2.5% DSS supply in drinking water for 5 days except for the control group. Colitis-associated DAI scores were significantly (p < 0.05) attenuated in HEAT and LYSOZYME group. The HEAT group exhibited significantly (p < 0.05) lower colonic tissue damage score compared to the DSS group. Furthermore, HEAT and LYSOZYME groups showed significantly (p < 0.05) higher colonic expressions of toll-like receptor (TLR) 6 and intestinal junction protein E-cadherin and occludin compared to the DSS group. LYSOZYME group showed significantly (p < 0.05) lower colonic expressions of Th2 cell-associated pro-inflammatory molecules, namely GATA3 and IL-4, and higher expression of anti-inflammatory NLRP6 and IL-18 compared to the DSS group. Also, HEAT group exhibited significantly (p < 0.05) lower colonic p-IκBα expression compared to the DSS group, while COX-2 expression was significantly (p < 0.05) suppressed by both paraprobiotics supplementation. Paraprobiotics significantly altered the composition of the intestinal microbiota. CONCLUSION: Paraprobiotic L. acidophilus PIN7 ameliorated DSS-induced colitis by regulating immune-modulatory TLR6 signalling and gut microbiota composition. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests paraprobiotic L. acidophilus PIN7 are superior candidates to prevent intestinal inflammation associated with dysregulated immune responses.

New glucogenesis inhibition model based on complete α-glucosidases from rat intestinal tissues validated with various types of natural and pharmaceutical inhibitors.

BACKGROUND: Inhibition of intestinal α-glucosidases from rat intestinal acetone powder (RIAP) has been widely used in research focused on regulating glucogenesis to be applied as a strategy to control obesity and type II diabetes. However, the crude extract has different compositions of α-glucosidases than a complete RIAP suspension due to enzymes anchored on the intestinal tissues after the extraction. Here, the inhibitory effects of different pharmaceutical and food-grade inhibitors on the enzymes in the RIAP suspension were investigated. RESULTS: Instead of crude extracts from RIAP, the RIAP suspension without the extraction process was applied to optimize the α-glucosidase inhibitory model by pharmaceutical/natural inhibitors. The results clearly showed that the half-maximal inhibitory concentration ratios of four individual α-glucosidases by various inhibitors were different between the RIAP suspension and the crude extract. In particular, isomaltase from the RIAP suspension required more inhibitors than the crude extraction did, as this enzyme is still anchored to the remaining intestinal tissue from the extraction process. CONCLUSION: The crude extract from RIAP contains only a portion of the enzymes, which poses limitations for determining the precise inhibitory properties by various types of enzyme inhibitors. On the contrary, an in vitro assay with RIAP suspension that has all the α-glucosidases is a more suitable method for determining digestibility of glycemic carbohydrates. This new approach can be applied to the development of natural/synthetic α-glucosidase inhibitors to attenuate the postprandial glycemic response more accurately. © 2022 Society of Chemical Industry.

Biocatalytic Fabrication of α-Glucan-Coated Porous Starch Granules by Amylolytic and Glucan-Synthesizing Enzymes as a Target-Specific Delivery Carrier.

In this study, we created biocatalytically coated porous starch granules (PSGs) using amylosucrase from Neisseria polysaccharea to apply them as an encapsulant for target-specific delivery. Field-emission scanning electron and confocal laser scanning microscopic images showed that the PSGs were completely concealed by the α-glucan coating layer. This carbohydrate-based encapsulant displayed higher amount of resistant glucan contents due to the elongated chains of the glucan coating, resulting in lower digestibility of these PSGs in simulated digestive fluid systems. Among the various PSGs evaluated, the highest loading efficiency for the bioactive molecule crocin was observed with the ß-amylase-induced PSGs (ß-PSGs) that had the smallest nanosize pores. Furthermore, α-glucan-coated ß-PSGs showed the highest capacity to preserve the loaded crocin when incubated in simulated digestive fluids. This suggests that the α-glucan-coated ß-PSGs can potentially be used for the delayed release of the core material in the upper region of the gastrointestinal tract. Therefore, this system can be potentially utilized as an effective carrier for colon-specific delivery, and the release of the bioactive compound can be triggered by beneficial intestinal microbiota.

Structural Analysis of Gluco-Oligosaccharides Produced by Leuconostoc lactis and Their Prebiotic Effect.

Leuconostoc lactis CCK940, which exhibits glycosyltransferase activity, produces oligosaccharides using sucrose and maltose as donor and receptor molecules, respectively. The oligosaccharides produced were purified by Bio-gel P2 chromatography and the purified oligosaccharides (CCK-oligosaccharides) consisted of only glucose. 1H-NMR analysis revealed that the CCK-oligosaccharides were composed of 77.6% α-1,6 and 22.4% α-1,4 glycosidic linkages, and the molecular weight of the CCK-oligosaccharides was found to be 9.42 × 102 Da. To determine the prebiotic effect of the CCK-oligosaccharides, various carbon sources were added in modified media. Growth of six probiotic strains, Lactobacillus casei, L. pentosus, L. plantarum, Weissella cibaria, Bifidobacterim animalis, and Saccharomyces cerevisiae, was better when the CCK-oligosaccharides were used as the sole carbon source compared to fructo-oligosaccharides, which are widely used as prebiotics. These results showed that the CCK-oligosaccharides produced from Leu. lactis CCK940 could serve as good candidates for novel prebiotics.

Maltase Has Most Versatile α-Hydrolytic Activity Among the Mucosal α-Glucosidases of the Small Intestine.

Complete digestion of the glycemic carbohydrates to glucose takes place through the combined action of the 4 mucosal α-glucosidases (maltase-glucoamylase and sucrase-isomaltase) in the small intestine. Maltase digests α-1,2- and α-1,3-disaccharides better than the other α-glucosidases, and has, as well, the capability to effectively hydrolyze α-1,4 and α-1,6 linkages that form the major backbone of a starch molecule. This broad hydrolytic activity on α-linkages makes it an enzyme that has the most versatile α-hydrolytic activity among the 4mucosal α-glucosidases. The slowly digestible properties of the unusual linkages from this research suggest the development of new glycemic oligosaccharides which will be hydrolyzed slowly, compared to α-1,4 linkages, for modulating the postprandial glycemic response. In addition, using mammalian mucosal α-glucosidases is a better fit to characterize carbohydrate digestion properties, compared to fungal amyloglucosidase which is currently applied in in vitro assays.

Altering the Structure of Carbohydrate Storage Granules in the Cyanobacterium Synechocystis sp. Strain PCC 6803 through Branching-Enzyme Truncations.

UNLABELLED: Carbohydrate storage is an important element of metabolism in cyanobacteria and in the chloroplasts of plants. Understanding how to manipulate the metabolism and storage of carbohydrate is also an important factor toward harnessing cyanobacteria for energy production. While most cyanobacteria produce glycogen, some have been found to accumulate polysaccharides in the form of water-insoluble α-glucan similar to amylopectin. Notably, this alternative form, termed "semi-amylopectin," forms in cyanobacterial species harboring three branching-enzyme (BE) homologs, designated BE1, BE2, and BE3. In this study, mutagenesis of the branching genes found in Synechocystis sp. strain PCC 6803 was performed in order to characterize their possible impact on polysaccharide storage granule morphology. N-terminal truncations were made to the native BE gene of Synechocystis sp. PCC 6803. In addition, one of the two native debranching enzyme genes was replaced with a heterologous debranching enzyme gene from a semi-amylopectin-forming strain. Growth and glycogen content of mutant strains did not significantly differ from those of the wild type, and ultrastructure analysis revealed only slight changes to granule morphology. However, analysis of chain length distribution by anion-exchange chromatography revealed modest changes to the branched-chain length profile. The resulting glycogen shared structure characteristics similar to that of granules isolated from semi-amylopectin-producing strains. IMPORTANCE: This study is the first to investigate the impact of branching-enzyme truncations on the structure of storage carbohydrates in cyanobacteria. The results of this study are an important contribution toward understanding the relationship between the enzymatic repertoire of a cyanobacterial species and the morphology of its storage carbohydrates.

Human α-amylase present in lower-genital-tract mucosal fluid processes glycogen to support vaginal colonization by Lactobacillus.

Lactobacillus colonization of the lower female genital tract provides protection from the acquisition of sexually transmitted diseases, including human immunodeficiency virus, and from adverse pregnancy outcomes. While glycogen in vaginal epithelium is thought to support Lactobacillus colonization in vivo, many Lactobacillus isolates cannot utilize glycogen in vitro. This study investigated how glycogen could be utilized by vaginal lactobacilli in the genital tract. Several Lactobacillus isolates were confirmed to not grow in glycogen, but did grow in glycogen-breakdown products, including maltose, maltotriose, maltopentaose, maltodextrins, and glycogen treated with salivary α-amylase. A temperature-dependent glycogen-degrading activity was detected in genital fluids that correlated with levels of α-amylase. Treatment of glycogen with genital fluids resulted in production of maltose, maltotriose, and maltotetraose, the major products of α-amylase digestion. These studies show that human α-amylase is present in the female lower genital tract and elucidates how epithelial glycogen can support Lactobacillus colonization in the genital tract.

Characterization of green banana starch from "Songkibab" species cultivated in the southern part of Korea.

Resistant starch (RS) has advantages for regulating the colon health as prebiotics and dietary fibers, and green banana has interested due to containing high amounts of RS. Here, the structural, physicochemical, and digestible characteristics of green banana starch from newly bred Songkibab (SB) were determined to evaluate its suitability for application as a new crop in response to global warming and for obtaining genetic diversity. SB starch has structural similarities to the Cavendish (CD) banana, which is widely consumed in Southeast Asia, in its ratio of B3-chains (in high amounts), flattened shapes of smooth surfaces, and B-type crystallinity. Physiochemically, SB shows comparable swelling power, amylose content, and viscosity pattern but a higher RS content. Conclusively, this study suggests that SB banana may be a good resource for replacing CD species with novel varieties in East Asia because of the high degree of similarity in the various characteristics. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01331-z.

α-Glucan-type exopolysaccharides with varied linkage patterns: Mitigating post-prandial glucose spike and prolonging the glycemic response.

Microbial exopolysaccharides (EPSs) are traditionally known as prebiotics that foster colon health by serving as microbiota nutrients, while remaining undigested in the small intestine. However, recent findings suggest that α-glucan structures in EPS, with their varied α-linkage types, can be hydrolyzed by mammalian α-glucosidases at differing rates. This study explores α-glucan-type EPSs, including dextran, alternan, and reuteran, assessing their digestive properties both in vitro and in vivo. Notably, while fungal amyloglucosidase - a common in vitro tool for carbohydrate digestibility analysis - shows limited efficacy in breaking down these structures, mammalian intestinal α-glucosidases can partially degrade them into glucose, albeit slowly. In vivo experiments with mice revealed that various EPSs elicited a significantly lower glycemic response (p < 0.05) than glucose, indicating their nature as carbohydrates that are digested slowly. This leads to the conclusion that different α-glucan-type EPSs may serve as ingredients that attenuate post-prandial glycemic responses. Furthermore, rather than serving as mere dietary fibers, they hold the potential for blood glucose regulation, offering new avenues for managing obesity, Type 2 diabetes, and other related-chronic diseases.

The branching ratio of enzymatically synthesized α-glucans impacts microbiome and metabolic outcomes of in vitro fecal fermentation.

The aim of this study was to evaluate the impacts of enzymatically synthesized α-glucans possessing α-1,4- and α-1,6-glucose linkages, and varying in branching ratio, on colonic microbiota composition and metabolic function. Four different α-glucans varying in branching ratio were synthesized by amylosucrase from Neisseria polysaccharea and glycogen branching enzyme from Rhodothermus obamensis. The branching ratios were found to range from 0 % to 2.8 % using GC/MS. In vitro fecal fermentation analyses (n = 8) revealed that the branching ratio dictates the short-chain fatty acid (SCFA) generation by fecal microbiota. Specifically, slightly branched (0.49 %) α-glucan resulted in generation of significantly (P < 0.05) higher amounts of propionate, compared to more-branched counterparts. In addition, the amount of butyrate generated from this α-glucan was statistically (P > 0.05) indistinguishable than those observed in resistant starches. 16S rRNA sequencing revealed that enzymatically synthesized α-glucans stimulated Lachnospiraceae and Ruminococcus related OTUs. Overall, the results demonstrated metabolic function of colonic microbiota can be manipulated by altering the branching ratio of enzymatically synthesized α-glucans, providing insights into specific structure-function relationships between dietary fibers and the colonic microbiome. Furthermore, the slightly branched α-glucans could be used as functional carbohydrates to stimulate the beneficial microbiota and SCFAs in the colon.

Undaria pinnatifida ameliorates nasal inflammation by inhibiting eosinophil and mast cell activation and modulating the NF-κB/MAPKs signaling pathway.

BACKGROUND: Allergic rhinitis (AR) is the most prevalent form of atopic disease. Undaria pinnatifida has potent antioxidative, antidiabetic, and anti-inflammatory properties. AIMS: We investigated the immunomodulatory effect of Undaria pinnatifida extract (UPE) on allergic inflammation in an AR mouse model. MATERIALS & METHODS: Mice were sensitized and intranasally challenged with ovalbumin (OVA), and the Th1/Th2 and Th17/Treg-related cytokines and histopathology were exanimated after UPE treatments. Enzyme-linked immunosorbent assay was performed using serum samples and NALF to detect OVA-specific immunoglobulins and inflammatory cytokines. Mitogen-activated protein kinases (MAPKs) were measured by western blotting analysis, and an in vitro study measured mast cell activation induced by compound 48/80. RESULTS: After UPE treatment, nasal and lung allergy symptoms, nasal mucosal swelling, and goblet cell hyperplasia were ameliorated. Oral UPE regulated the balance of Th1/Th2 and Th17/Treg cell differentiation in AR mice in a dose-dependent manner. In addition, UPE attenuated the migration of eosinophils and mast cells to the nasal mucosa by suppressing nuclear factor kappa B (NF-κB)/MAPKs. The levels of anti-OVA IgE and IgG1 were also decreased. DISCUSSION: UPE inhibited inflammation by regulating the NF-κB/MAPKs signaling pathway and supressing the activation of critical immune cells such as eosinophils and mast cells. CONCLUSION: UPE may have therapeutic potential for AR.

Efficient biotransformation of naringenin to naringenin α-glucoside, a novel α-glucosidase inhibitor, by amylosucrase from Deinococcus wulumuquiensis.

Amylosucrase (ASase) efficiently biosynthesizes α-glucoside using flavonoids as acceptor molecules and sucrose as a donor molecule. Here, ASase from Deinococcus wulumuqiensis (DwAS) biosynthesized more naringenin α-glucoside (NαG) with sucrose and naringenin as donor and acceptor molecules, respectively, than other ASases from Deinococcus sp. The biotransformation rate of DwAS to NαG was 21.3% compared to 7.1-16.2% for other ASases. Docking simulations showed that the active site of DwAS was more accessible to naringenin than those of others. The 217th valine in DwAS corresponded to the 221st isoleucine in Deinococcus geothermalis AS (DgAS), and the isoleucine possibly prevented naringenin from accessing the active site. The DwAS-V217I mutant had a significantly lower biosynthetic rate of NαG than DwAS. The kcat/Km value of DwAS with naringenin as the donor was significantly higher than that of DgAS and DwAS-V217I. In addition, NαG inhibited human intestinal α-glucosidase more efficiently than naringenin.

Starch source influences dietary glucose generation at the mucosal α-glucosidase level.

The quality of starch digestion, related to the rate and extent of release of dietary glucose, is associated with glycemia-related problems such as diabetes and other metabolic syndrome conditions. Here, we found that the rate of glucose generation from starch is unexpectedly associated with mucosal α-glucosidases and not just α-amylase. This understanding could lead to a new approach to regulate the glycemic response and glucose-related physiologic responses in the human body. There are six digestive enzymes for starch: salivary and pancreatic α-amylases and four mucosal α-glucosidases, including N- and C-terminal subunits of both maltase-glucoamylase and sucrase-isomaltase. Only the mucosal α-glucosidases provide the final hydrolytic activities to produce substantial free glucose. We report here the unique and shared roles of the individual α-glucosidases for α-glucans persisting after starch is extensively hydrolyzed by α-amylase (to produce α-limit dextrins (α-LDx)). All four α-glucosidases share digestion of linear regions of α-LDx, and three can hydrolyze branched fractions. The α-LDx, which were derived from different maize cultivars, were not all equally digested, revealing that the starch source influences glucose generation at the mucosal α-glucosidase level. We further discovered a fraction of α-LDx that was resistant to the extensive digestion by the mucosal α-glucosidases. Our study further challenges the conventional view that α-amylase is the only rate-determining enzyme involved in starch digestion and better defines the roles of individual and collective mucosal α-glucosidases. Strategies to control the rate of glucogenesis at the mucosal level could lead to regulation of the glycemic response and improved glucose management in the human body.

Modulation of starch digestion for slow glucose release through "toggling" of activities of mucosal α-glucosidases.

Starch digestion involves the breakdown by α-amylase to small linear and branched malto-oligosaccharides, which are in turn hydrolyzed to glucose by the mucosal α-glucosidases, maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI). MGAM and SI are anchored to the small intestinal brush-border epithelial cells, and each contains a catalytic N- and C-terminal subunit. All four subunits have α-1,4-exohydrolytic glucosidase activity, and the SI N-terminal subunit has an additional exo-debranching activity on the α-1,6-linkage. Inhibition of α-amylase and/or α-glucosidases is a strategy for treatment of type 2 diabetes. We illustrate here the concept of "toggling": differential inhibition of subunits to examine more refined control of glucogenesis of the α-amylolyzed starch malto-oligosaccharides with the aim of slow glucose delivery. Recombinant MGAM and SI subunits were individually assayed with α-amylolyzed waxy corn starch, consisting mainly of maltose, maltotriose, and branched α-limit dextrins, as substrate in the presence of four different inhibitors: acarbose and three sulfonium ion compounds. The IC(50) values show that the four α-glucosidase subunits could be differentially inhibited. The results support the prospect of controlling starch digestion rates to induce slow glucose release through the toggling of activities of the mucosal α-glucosidases by selective enzyme inhibition. This approach could also be used to probe associated metabolic diseases.

Slow digestion properties of long-sized isomaltooligosaccharides synthesized by a transglucosidase from Thermoanaerobacter thermocopriae.

Isomaltooligosaccharides (IMOs) are widely used as prebiotic ingredients that promote colon health; however, recent studies revealed that these are slowly hydrolyzed to glucose within the small intestine. Here, novel α-glucans with a higher number of α-1,6 linkages were synthesized from maltodextrins using the Thermoanaerobacter thermocopriae-derived transglucosidase (TtTG) to decrease susceptibility to hydrolysis and improve slow digestion properties. The synthesized long-sized IMOs (l-IMOs; 70.1% of α-1,6 linkages), comprising 10-12 glucosyl units, exhibited slow hydrolysis to glucose when compared to commercial IMOs under treatment with mammalian α-glucosidase level. In male mice, the ingestion of l-IMOs significantly decreased the post-prandial glycemic response compared to other samples (p < 0.05). Therefore, enzymatically synthesized l-IMOs can be applied as functional ingredients for the modulation of blood glucose homeostasis in obesity, Type 2 diabetes, and other chronic diseases.

Production of highly branched α-limit dextrins with enhanced slow digestibility by various glycogen-branching enzymes.

α-Limit dextrins (α-LDx) are slowly digestible carbohydrates that attenuate postprandial glycemic response and trigger the secretion of satiety-related hormones. In this study, more highly branched α-LDx were enzymatically synthesized to enhance the slowly digestible property by various origins of glycogen branching enzyme (GBE), which catalyzes the transglycosylation to form α-1,6 branching points after cleaving α-1,4 linkages. Results showed that the proportion of branched α-LDx in starch molecules increased around 2.2-8.1 % compared to α-LDx from starch without GBE treatment as the ratio of α-1,6 linkages increased after different types of GBE treatments. Furthermore, the enzymatic increment of branching points enhanced the slowly digestible properties of α-LDx at the mammalian α-glucosidase level by 17.3-28.5 %, although the rates of glucose generation were different depending on the source of GBE treatment. Thus, the highly branched α-LDx with a higher amount of α-1,6 linkages and a higher molecular weight can be applied as a functional ingredient to deliver glucose throughout the entire small intestine without a glycemic spike which has the potential to control metabolic diseases such as obesity and type 2 diabetes.

Macromolecular α-glucans with α-1,3/α-1,4 branching structures produced using dual glycosyltransferases: Elucidation of physicochemical and slowly digestible properties.

Amylosucrase from Neisseria polysaccharea (NpAS) produces the linear amylose-like α-glucans by the elongation property from sucrose, and 4,3-α-glucanotransferase from Lactobacillus fermentum NCC 2970 (4,3-αGT) newly synthesizes the α-1,3 linkages after cleaving the α-1,4 linkages by the glycosyltransferring property. This study focused on the synthesis of high molecular α-1,3/α-1,4-linked glucans by combining NpAS and 4,3-αGT and analyzed their structural and digestive properties. The enzymatically synthesized α-glucans have a molecular weight of >1.6 × 107 g mol-1, and the α-4,3 branching ratios on the structures increased as the amount of 4,3-αGT increased. The synthesized α-glucans were hydrolyzed to linear maltooligosaccharides and α-4,3 branched α-limit dextrins (α-LDx) by human pancreatic α-amylase, and the amounts of produced α-LDx were increased depending on the ratio of synthesized α-1,3 linkages. In addition, approximately 80 % of the synthesized products were partially hydrolyzed by mammalian α-glucosidases, and the glucose generation rates decelerated as the amounts of α-1,3 linkages increased. In conclusion, new types of α-glucans with α-1,4 and α-1,3 linkages were successfully synthesized by a dual enzyme reaction. These can be utilized as slowly digestible and prebiotic ingredients in the gastrointestinal tract due to their novel linkage patterns and high molecular weights.

Modulation of gut microbiota by rice starch enzymatically modified using amylosucrase from Deinococcus geothermalis.

Amylosucrase can increase the amount of resistant starch (RS) in starch by transferring glucose from sucrose to amylopectin. Here, rice starch was modified using amylosucrase from Deinococcus geothermalis (DgAS). DgAS-modified rice starch (DMRS) increased the side-chain length of amylopectin and appeared in the form of B-type crystals. In vitro digestion analyses revealed that DMRS had a higher RS contents and lower digestion rate than native rice starch. When high-fat diet (HFD)-induced C57BL/6 mice were orally administered DMRS, body weight and white fat tissues of DMRS-fed HFD mice were not significantly different. However, serum leptin and glucose levels were significantly decreased and serum glucagon like peptide-1was increased in these mice. The cecal microbiome in DMRS-fed HFD mice was identified to investigate the role of DMRS in gut microbiota regulation. DMRS supplementation increased the relative abundance of Bacteroides, Faecalibaculum, and Ruminococcus in mouse gut microbiota. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01238-1.

Structural analysis and prebiotic activity of exopolysaccharide produced by probiotic strain Bifidobacterium bifidum EPS DA-LAIM.

Exopolysaccharide (EPS)-producing Bifidobacterium bifidum EPS DA-LAIM was isolated from healthy human feces, the structure of purified EPS from the strain was analyzed, and its prebiotic activity was evaluated. The EPS from B. bifidum EPS DA-LAIM is a glucomannan-type heteropolysaccharide with a molecular weight of 407-1007 kDa, and its structure comprises 2-mannosyl, 6-mannosyl, and 2,6-mannosyl residues. The purified EPS promoted the growth of representative lactic acid bacteria and bifidobacterial strains. Bifidobacterium bifidum EPS DA-LAIM increased nitric oxide production in RAW 264.7 macrophage cells, indicating its immunostimulatory activity. Bifidobacterium bifidum EPS DA-LAIM also exhibited high gastrointestinal tract tolerance, gut adhesion ability, and antioxidant activity. These results suggest that EPS from B. bifidum EPS DA-LAIM is a potentially useful prebiotic material, and B. bifidum EPS DA-LAIM could be applied as a probiotic candidate. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01213-w.

Effect of highly branched α-glucans synthesized by dual glycosyltransferases on the glucose release rate.

Increasing α-1,6 linkages in starch molecules generates a large amount of α-limit dextrins (α-LDx) during α-amylolysis, which decelerate the release of glucose at the intestinal α-glucosidase level. This study synthesized highly branched α-glucans from sucrose using Neisseria polysaccharea amylosucrase and Rhodothermus obamensis glycogen branching enzyme to enhance those of slowly digestible property. The synthesized α-glucans (Mw: 1.7-4.9 × 107 g mol-1) were mainly composed of α-1,4 linkages and large proportions of α-1,6 linkages (7.5%-9.9%). After treating the enzymatically synthesized α-glucans with the human α-amylase, the quantity of branched α-LDx (36.2%-46.7%) observed was higher than that for amylopectin (26.8%) and oyster glycogen (29.1%). When the synthetic α-glucans were hydrolyzed by mammalin α-glucosidases, the glucose generation rate decreased because the amount of embedded branched α-LDx increased. Therefore, the macro-sized branched α-glucans with high α-LDx has the potential to be used as slowly digestible material to attenuate postprandial glycemic response.