To compete for the dynamic stream of nutrients flowing into their ecosystem, colonic bacteria must respond rapidly to new resources and then catabolize them efficiently once they are detected. The Bacteroides thetaiotaomicron starch utilization system (Sus) is a model for nutrient acquisition by symbiotic gut bacteria, which harbor thousands of related Sus-like systems. Structural investigation of the four Sus outer membrane proteins (SusD, -E, -F, and -G) revealed that they contain a total of eight starch-binding sites that we demonstrated, using genetic and biochemical approaches, to play distinct roles in starch metabolism in vitro and in vivo in gnotobiotic mice. SusD, whose homologs are abundant in the human microbiome, is critical for the initial sensing of available starch, allowing sus transcriptional activation at much lower concentrations than without this function. In contrast, seven additional binding sites across SusE, -F, and -G are dispensable for sus activation. However, they optimize the rate of growth on starch in a manner dependent on the expression of the bacterial polysaccharide capsule, suggesting that they have evolved to offset the diffusion barrier created by this structure. These findings demonstrate how proteins with similar biochemical behavior can serve orthogonal functions during different stages of cellular adaptation to nutrients. Finally, we demonstrated in gnotobiotic mice fed a starch-rich diet that the Sus binding sites confer a competitive advantage to B. thetaiotaomicron in vivo in a manner that is dependent on other colonizing microbes. This study reveals how numerically dominant families of carbohydrate-binding proteins in the human microbiome fulfill separate and sometimes cooperative roles to optimize gut commensal bacteria for nutrient acquisition.

For digestion of starch in humans, α-amylase first hydrolyzes starch molecules to produce α-limit dextrins, followed by complete hydrolysis to glucose by the mucosal α-glucosidases in the small intestine. It is known that α-1,6 linkages in starch are hydrolyzed at a lower rate than are α-1,4 linkages. Here, to create designed slowly digestible carbohydrates, the structure of waxy corn starch (WCS) was modified using a known branching enzyme alone (BE) and an in combination with β-amylase (BA) to increase further the α-1,6 branching ratio. The digestibility of the enzymatically synthesized products was investigated using α-amylase and four recombinant mammalian mucosal α-glucosidases. Enzyme-modified products (BE-WCS and BEBA-WCS) had increased percentage of α-1,6 linkages (WCS: 5.3%, BE-WCS: 7.1%, and BEBA-WCS: 12.9%), decreased weight-average molecular weight (WCS: 1.73×108 Da, BE-WCS: 2.76×105 Da, and BEBA-WCS 1.62×105 Da), and changes in linear chain distributions (WCS: 21.6, BE-WCS: 16.9, BEBA-WCS: 12.2 DPw). Hydrolysis by human pancreatic α-amylase resulted in an increase in the amount of branched α-limit dextrin from 26.8% (WCS) to 56.8% (BEBA-WCS). The α-amylolyzed samples were hydrolyzed by the individual α-glucosidases (100 U) and glucogenesis decreased with all as the branching ratio increased. This is the first report showing that hydrolysis rate of the mammalian mucosal α-glucosidases is limited by the amount of branched α-limit dextrin. When enzyme-treated materials were gavaged to rats, the level of postprandial blood glucose at 60 min from BEBA-WCS was significantly higher than for WCS or BE-WCS. Thus, highly branched glucan structures modified by BE and BA had a comparably slow digesting property both in vitro and in vivo. Such highly branched α-glucans show promise as a food ingredient to control postprandial glucose levels and to attain extended glucose release.

For digestion of starch, α‐amylase first hydrolyzes the starch structure to α‐limit dextrins (αLDx's). Complete hydrolysis to glucose then takes place through the combined action of mucosal maltase‐glucoamylase (MGAM) and sucrase‐isomaltase (SI), which have two subunits each (N‐ and C‐ terminal). In this study, we applied the concept of “toggling” through differential inhibition of subunits to examine control of glucogenesis from αLDx's with the aim of attaining slow glucose delivery to the body. Mammalian recombinant MGAM and SI subunits were individually reacted with αLDx's with varying concentrations of acarbose, a well known α‐glucosidase inhibitor. Released glucose amounts were analyzed for hydrolysis activity. Notably, results showed selective inhibitory effect on C‐terminal subunits by acarbose for starch digestion. Furthermore, Ki values of C‐terminal subunits were lower than human α‐amylases which can be applied that certain amount of acarbose had inhibitory effect on the hydrolysis of αLDx's by C‐terminal subunits but not on α‐amylase for moderating glucogenesis. This result supports the concept of controlling starch digestion rate for slow glucose release through the “toggling” of activities of the mucosal α‐glucosidases by selective enzyme inhibition. Conceivably this approach could be used to treat Type II diabetes by extending postprandial blood glucose delivery to the body, and may apply as well to other metabolic syndrome‐associated diseases and conditions. This research was supported from Whistler Center for Carbohydrate Research and USDA/AFRI.

Turanose is a sucrose isomer naturally existing in honey and a promising functional sweetener due to its low glycemic response. In this study, the extrinsic fructose effect on turanose productivity was examined in Neisseria amylosucrase reaction. Turanose was produced, by increasing the amount of extrinsic fructose as a reaction modulator, with high concentration of sucrose substrate, which resulted in 73.7% of production yield. In physiological functionality test, lipid accumulation in 3T3-L1 preadipocytes in the presence of high amounts of pure glucose was attenuated by turanose substitution in a dose-dependent manner. Turanose treatments at concentrations representing 50%, 75%, and 100% of total glucose concentration in cell media significantly reduced lipid accumulation by 18%, 35%, and 72%, respectively, as compared to controls. This result suggested that turanose had a positive role in controlling adipogenesis, and enzymatic process of turanose production has a potential to develop a functional food ingredient for controlling obesity and related chronic diseases.

Enzymatic hydrolysis and self-assembly are considered promising methods for preparation of starch nanoparticles (SNPs) because they are environmentally friendly, and time- and cost-effective. These methods are based on the self-assembly of short-chain glucans released from the α-1,6 bonds in amylopectin. Since their discovery, many studies have described the structural and physicochemical properties of self-assembled SNPs. Self-assembled SNPs can be prepared by two methods: using only the soluble portion containing the short-chain glucans, or using the whole hydrolyzate including both insoluble and soluble fractions. Although the structural and physical properties of self-assembled SNPs can be attributed to the composition of the hydrolyzates that participate in self-assembly, this aspect has not yet been discussed. This review focuses on SNPs self-assembled with only soluble short-chain glucans and addresses their characteristics, including formation mechanisms as well as structural and physicochemical properties, compared with SNPs prepared with total hydrolyzates.

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