Microbial enzymatic production and applications of short-chain fructooligosaccharides and inulooligosaccharides: recent advances and current perspectives.

The industrial production of short-chain fructooligosaccharides (FOS) and inulooligosaccharides is expanding rapidly due to the pharmaceutical importance of these compounds. These compounds, concisely termed prebiotics, have biofunctional properties and hence health benefits if consumed in recommended dosages. Prebiotics can be produced enzymatically from sucrose elongation or via enzymatic hydrolysis of inulin by exoinulinases and endoinulinases acting alone or synergistically. Exoinulinases cleave the non-reducing ß-(2, 1) end of inulin-releasing fructose while endoinulinases act on the internal linkages randomly to release inulotrioses (F3), inulotetraoses (F4) and inulopentaoses (F5) as major products. Fructosyltransferases act by cleaving a sucrose molecule and then transferring the liberated fructose molecule to an acceptor molecule such as sucrose or another oligosaccharide to elongate the short-chain fructooligosaccharide. The FOS produced by the action of fructosyltransferases are 1-kestose (GF2), nystose (GF3) and fructofuranosyl nystose (GF4). The production of high yields of oligosaccharides of specific chain length from simple raw materials such as inulin and sucrose is a technical challenge. This paper critically explores recent research trends in the production and application of short-chain oligosaccharides. Inulin and enzyme sources for the production of prebiotics are discussed. The mechanism of FOS chain elongation and also the health benefits associated with prebiotics consumption are discussed in detail.

Inulin-type prebiotics--a review: part 1.

This article is part 1 of a two-part review of inulin-type prebiotics. Prebiotics are a category of nutritional compounds grouped together by the ability to promote the growth of specific beneficial (probiotic) gut bacteria. Inulin-type prebiotics contain fructans of the inulin-type. Fructans are a category of nutritional compounds that encompasses naturally occurring plant oligo- and polysaccharides in which one or more fructosyl-fructose linkages comprise the majority of glycosidic bonds. To be inulin-type a fructan must have beta (2(1) fructosyl-fructose glycosidic bonds, which gives inulin its unique structural and physiological properties, allowing it to resist enzymatic hydrolysis by human salivary and small intestinal digestive enzymes. Inulin-type prebiotics include fructooligosaccharides (FOS), oligofructose, and inulin - terms that have been used inconsistently in both the scientific literature and in food applications. Commercially available inulin-type prebiotics can be extracted from food (typically chicory root) or synthesized from a more fundamental molecule (typically sucrose). Depending on the starting source and degree of processing, inulin-type prebiotics can be produced with very different chemical compositions. Some inulin-type prebiotics are relatively high in free sugars (the monosaccharides fructose and glucose and the disaccharide sucrose), while others have most or all free sugars removed. Processing can also result in mixes consisting exclusively of inulin-type oligosaccharides, polysaccharides, or both. Because inulin, oligofructose, and FOS resist enzymatic digestion in the upper gastrointestinal tract, they reach the colon virtually intact where they undergo bacterial fermentation. All inulin-type prebiotics are bifidogenic - stimulating the growth of Bifidobacteria species. The effects they have on other gut organisms are less consistent. A minimal dose of inulin-type prebiotic appears to be needed to produce a bifidogenic effect. However, intra-individual response to an identical dose of the same inulin-type prebiotic, in terms of stimulation of total number of Bifidobacteria and individual Bifidobacteria species, can be variable. Research on therapeutic uses of inulin-type prebiotics will be covered in part 2 of this review.

The effect of fructan on the phospholipid organization in the dry state.

Fructans, a family of oligo- and polyfructoses, are implicated to play a drought-protecting role in plants. Inulin-type fructan is able to preserve the membrane barrier during dehydration. However, whether other fructans would be able to perform this function is unknown. In addition, almost nothing is known about the organization of these systems, which could give insight into the protective mechanism. To get insight into these questions the effect of different fructans on phosphatidylcholine-based model systems under conditions of dehydration was analyzed. Using a vesicle leakage assay, it was found that both levan- and inulin-type fructans protected the membrane barrier. This suggests that fructans in general would be able to protect the membrane barrier function. Furthermore, both fructan-types inhibited vesicle fusion to a large extent as measured using a lipid-mixing assay. Using x-ray diffraction, it was found that in the presence of both inulin- and levan-type fructans the lamellar repeat distance increased considerably. From this it was concluded that fructans are present between the lipid bilayers during drying. Furthermore, they stabilize the L(alpha) phase. In contrast to fructans, dextran did not increase the lamellar repeat distance and it even promoted L(beta) phase formation. These data support the hypothesis that fructans can have a membrane-protecting role during dehydration, and give insight into the mechanism of protection.

Structural requirements of the fructan-lipid interaction.

Fructans are a group of fructose-based oligo- and polysaccharides. They are proposed to be involved in membrane protection of plants during dehydration. In accordance with this hypothesis, they show an interaction with hydrated lipid model systems. However, the structural requirements for this interaction are not known both with respect to the fructans as to the lipids. To get insight into this matter, the interaction of several inulins and levan with lipids was investigated using a monomolecular lipid system or the MC 540 probe in a bilayer system. MD was used to get conformational information concerning the polysaccharides. It was found that levan-type fructan interacted comparably with model membranes composed of glyco- or phospholipids but showed a preference for lipids with a small headgroup. Furthermore, it was found that there was an inulin chain-length-dependent interaction with lipids. The results also suggested that inulin-type fructan had a more profound interaction with the membrane than levan-type fructan. MD simulations indicated that the favorable conformation for levan is a helix, whereas inulin tends to form random coil structures. This suggests that flexibility is an important determinant for the fructan-lipid interaction.