In November 2008, a group of scientists met at the 6th Meeting of the International Scientific Association of Probiotics and Prebiotics (ISAPP) in London, Ontario, Canada, to discuss the functionality of prebiotics. As a result of this, it was concluded that the prebiotic field is currently dominated by gastrointestinal events. However, in the future, it may be the case that other mixed microbial ecosystems may be modulated by a prebiotic approach, such as the oral cavity, skin and the urogenital tract. Therefore, a decision was taken to build upon the current prebiotic status and define a niche for 'dietary prebiotics'. This review is co-authored by the working group of ISAPP scientists and sets the background for defining a dietary prebiotic as ''a selectively fermented ingredient that results in specific changes in the composition and/or activity of the gastrointestinal microbiota, thus conferring benefit(s) upon host health''.

The objective of this study was to isolate and characterize the meat factor(s) that enhances nonheme iron bioavailability using various analytical and in vitro cell culture techniques. Nonheme iron bioavailability was measured via radiolabeled iron uptake or ferritin formation in Caco-2 cells. Fish haddock fillet was cooked and lyophilized to be used as the muscle tissue of choice because of its low intrinsic iron content. It was demonstrated that the low pH of the stomach (pH 2.0) was the primary factor responsible for initiating the enhancing effect of fish on nonheme iron uptake. Subsequently, cooked fish samples were titrated with HCl to pH 2.0 and incubated for 1 h without digestive enzymes to release the factor(s) from the fish. The supernatant of this acidic digest was then used as a starting material for the meat factor isolation procedures. Fractions generated through Sephadex G-25 size exclusion increased Caco-2 cell iron uptake approximately 9-fold. Subsequent chromatography of these fractions via C18 reverse-phase HPLC were conducted, and enhancing activity was observed only in the "injection peak." This observation coupled with protein measurement and amino acid composition analysis revealed that the active fractions contained negligible amounts of proteins or amino acids. Active fractions were highly enriched with carbohydrates. Subsequent chromatography via high performance anion exchange chromatography with pulsed amperometric detection yielded 3 active peaks that increased Caco-2 cell iron uptake 3.4- to 4.9-fold. Our results indicate that specific carbohydrates contribute to the enhancing effect of meat on iron uptake by the enterocyte. These carbohydrates may be oligosaccharides originating from glycosaminoglycans in the extracellular matrix of muscle tissue.

The three-dimensional structure of a complex between the pectate lyase C (PelC) R218K mutant and a plant cell wall fragment has been determined by x-ray diffraction techniques to a resolution of 2.2 A and refined to a crystallographic R factor of 18.6%. The oligosaccharide substrate, alpha-D-GalpA-([1-->4]-alpha-D-GalpA)3-(1-->4)-D-GalpA , is composed of five galacturonopyranose units (D-GalpA) linked by alpha-(1-->4) glycosidic bonds. PelC is secreted by the plant pathogen Erwinia chrysanthemi and degrades the pectate component of plant cell walls in soft rot diseases. The substrate has been trapped in crystals by using the inactive R218K mutant. Four of the five saccharide units of the substrate are well ordered and represent an atomic view of the pectate component in plant cell walls. The conformation of the pectate fragment is a mix of 21 and 31 right-handed helices. The substrate binds in a cleft, interacting primarily with positively charged groups: either lysine or arginine amino acids on PelC or the four Ca2+ ions found in the complex. The observed protein-oligosaccharide interactions provide a functional explanation for many of the invariant and conserved amino acids in the pectate lyase family of proteins. Because the R218K PelC-galacturonopentaose complex represents an intermediate in the reaction pathway, the structure also reveals important details regarding the enzymatic mechanism. Notably, the results suggest that an arginine, which is invariant in the pectate lyase superfamily, is the amino acid that initiates proton abstraction during the beta elimination cleavage of polygalacturonic acid.

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The survival of single strains of Bifidobacterium breve, Bifidobacterium longum, Lactobacillus acidophilus, and Lactobacillus reuteri was investigated in synbiotics that included 10 mg/ml of fructo-oligosaccharides, inulin and pectic-oligosaccharides in an alginate matrix under refrigerated (4 °C) aerobic storage conditions. When the matrices were cross-linked with calcium (45 mM), 102-103 cfu/ml of L. acidophilus and L. reuteri, and 0-103 cfu/ml of B. breve and B. longum survived refrigerated aerobic storage for 28 days. Following refrigerated storage, acetic (3-9 mM), butyric (0-2 mM), propionic (5-16 mM) and lactic acids (1-48 mM) were produced during the growth of probiotics in BHI broth at 37 °C, suggesting their metabolic activity after storage was stressed. When calcium cross-linking was not used in synbiotics, the matrix remained more gel-like after inoculation when compared to the calcium cross-linked matrix. At least 107 cfu/ml of probiotic bacteria survived after 21 days of storage within these gel-like alginate matrices. Significantly higher levels of B. breve, L. acidophilus and L. reuteri were obtained from the synbiotic matrices supplemented with fructo-oligosaccharides, inulin and pectic-oligosaccharides compared to alginate alone. B. longum survival was the same (~7 logs) in all gel-like synbiotic matrices. These results show that synbiotics protected probiotic bacteria and extended their shelf-life under refrigerated aerobic conditions. Synbiotics represent a viable delivery vehicle for health-promoting bacteria.