1-kestose and fungitetraose

The nutritional and therapeutic benefits of prebiotics have attracted the keen interest of consumers and food processing industry for their use as food ingredients. Fructo-oligosaccharides (FOS), new alternative sweeteners, constitute 1-kestose, nystose, and 1-beta-fructofuranosyl nystose produced from sucrose by the action of fructosyltransferase from plants, bacteria, yeast, and fungi. FOS has low caloric values, non-cariogenic properties, and help gut absorption of ions, decrease levels of lipids and cholesterol and bifidus-stimulating functionality. The purified linear fructose oligomers are added to various food products like cookies, yoghurt, infant milk products, desserts, and beverages due to their potential health benefits. This review is focused on the various aspects of biotechnological production, purification and potential applications of fructo-oligosaccharides.

Topics: Food Technology; Hexosyltransferases; Humans; Oligosaccharides; Prebiotics; Trisaccharides

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 β-(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.

Topics: Bacteria; Fructans; Fructose; Fungi; Glycoside Hydrolases; Hexosyltransferases; Inulin; Oligosaccharides; Prebiotics; Sucrose; Trisaccharides

The aim of the present study was to determine the effects of dietary administration of a fructooligosaccharide preparation rich in kestose and nestose on growth performance and gastrointestinal parameters in young turkeys. The kestose and nestose preparation was obtained through bioconversion of sucrose using fungi fructosyl transferase and contained in DM 39.9% of kestose, 17.6% of nystose, as well as 26.5% of glucose and 14.7% of sucrose. Three dietary levels of the sum of kestose and nystose (0.3, 0.6, and 1.2%) were fed to growing turkeys for 8 wk. When compared with the control treatment, addition of the kestose and nestose preparation had no effect on feed intake, feed conversion, and BW. The kestose and nestose-supplemented diet, especially the medium level of kestose and nystose, influenced microbial metabolism, especially in the ceca. Compared with the control group, the medium level of kestose and nestose decreased relative weight of gizzard (from 18.67 to 16.51 g/kg of BW) and weight of small intestine tissue (from 23.3 to 19.6 g/kg of BW) and increased weight of ceca digesta (from 3.51 to 4.77 g/kg of BW) as well as activities of microbial beta-glucosidase (an increase from 0.22 to 0.38 U/g) and alpha-galactosidase (an increase from 0.90 to 1.61 U/g), pH of digesta (a decrease from 6.13 to 5.79), concentration of NH3 (an increase from 0.60 to 0.98 mg/g), and concentration of total short-chain fatty acids (an increase from 81.1 to 107.7 micromol/g) in the cecal digesta. A high content of kestose and nestose in the diet caused a decrease in ileal and cecal pH (to 5.42 and 5.49, respectively).

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Cecum; Colon; Diet; Dietary Supplements; Dose-Response Relationship, Drug; Drug Therapy, Combination; Gastrointestinal Tract; Male; Oligosaccharides; Organ Size; Trisaccharides; Turkeys; Weight Gain

In this study, various levels of ultra-high pressure (UHP) were combined with the enzymatic synthesis of the fructooligosaccharide (FOS) using Pectinex Ultra SP-L and inulinase. The combination enhanced the FOS yields up to 2.5- and 1.5-fold, respectively, compared to atmospheric condition (0.1 MPa). However, the enzymatic reaction was dependent on the levels of pressure, the reaction times, and the initial sucrose concentrations. The combined UHP and inulinase showed that the maximum FOS yield (71.81%) was obtained under UHP at 200 MPa for 20 min with 300 g/L of initial sucrose as a substrate, while the FOS yield (57.13%) using Pectinex Ultra SP-L was obtained under UHP at 300 MPa for 15 min with 600 g/L of initial sucrose as a substrate. The FOS composition produced by Pectinex Ultra SP-L under the UHP was 1-kestose (GF

Topics: Aspergillus niger; Glycoside Hydrolases; Hydrolases; Hydrolysis; Industrial Microbiology; Oligosaccharides; Pressure; Sucrose; Trisaccharides

We report the production of the functional trisaccharide 1-kestose, O-β-D-fructofuranosyl-(2→1)-β-D-fructofuranosyl α-D-glucopyranoside, by β-fructofuranosidase from Aspergillus japonicus using sugar cane molasses as substrate. Sucrose in cane sugar molasses acted as a fructosyl donor and acceptor for the enzyme. The tetrasaccharide nystose, O-β-D-fructofuranosyl-(2→1)-β-D-fructofuranosyl-(2→1)-β-D-fructofuranosyl α-D-glucopyranoside, was produced from 1-kestose. Cane sugar molasses mixed with water provided a better substrate solution for β-fructofuranosidase compared to undiluted molasses due to the high concentration of product inhibitors such as glucose and fructose in molasses. The maximum concentration of 1-kestose obtained was 84.9 mg/ml and the maximum production efficiency was 32.3% after 24 h reaction at 40 °C. The maximum efficiency of combined fructo-oligosaccharide (1-kestose and nystose) production was 40.6%. 1-Kestose was therefore produced via a fructosyl-transfer reaction catalyzed by β-fructofuranosidase from A. japonicus.

Topics: Aspergillus; beta-Fructofuranosidase; Biocatalysis; Fungal Proteins; Kinetics; Molasses; Oligosaccharides; Saccharum; Trisaccharides

Fructooligosaccharides (FOS) are usually added in milk powder as a kind of prebiotic. Thus, quantitative analysis of the FOS is very important for the quality control of milk powder. In this study, a simple method for the simultaneous determination of three FOS components with degrees of polymerization (DP) 3-5 in milk powder was developed by high performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD). The samples were extracted with 50% (v/v) ethanol aqueous solution and defatted by an On Guard RP pretreatment column. The separation was performed on a CarboPac PA200 column by gradient elution using deionized water, 0. 2 mol/L NaOH solution and 0. 4 mol/L NaAc solution as the mobile phases. The flow rate was 0. 4 mL/min. The column temperature was 30 °C; and the injection volume was 25 µL. Good linear response was observed in the concentration range of 0.05-10 mg/L (r2 >0. 9993). The limits of quantification were 0. 02, 0. 005 and 0. 02 mg/L for 1-kestose, nystose and fructofuranosyl-nystose, respectively. The mean recoveries varied from 86. 0% to 114. 0% at three spiked levels of 0. 5, 1. 0 and 5. 0 mg/L. The short-chain fructooligosaccharides from inulooligosaccharides (IOS) were successfully separated by the developed HPAEC-PAD method. The method is simple, accurate, sensitive, and helpful for the quality control of milk powder.

Topics: Animals; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Milk; Oligosaccharides; Trisaccharides

Elevated [CO2 ] is suggested to mitigate the negative effects of water stress in plants; however responses vary among species. Fructans are recognised as protective compounds against drought and other stresses, as well as having a role as reserve carbohydrates. We analysed the combined effects of elevated [CO2 ] and water deficit on fructan metabolism in the Cerrado species Viguiera discolor Baker. Plants were cultivated for 18 days in open-top chambers (OTC) under ambient (∼380 ppm), and high (∼760 ppm) [CO2 ]. In each OTC, plants were submitted to three treatments: (i) daily watering (control), (ii) withholding water (WS) for 18 days and (iii) re-watering (RW) on day 11. Analyses were performed at time 0 and days 5, 8, 11, 15 and 18. High [CO2 ] increased photosynthesis in control plants and increased water use efficiency in WS plants. The decline in soil water content was more distinct in WS 760 (WS under 760 ppm), although the leaf and tuberous root water status was similar to WS 380 plants (WS under 380 ppm). Regarding fructan active enzymes, 1-SST activity decreased in WS plants in both CO2 concentrations, a result consistent with the decline in photosynthesis and, consequently, in substrate availability. Under WS and both [CO2 ] treatments, 1-FFT and 1-FEH seemed to act in combination to generate osmotically active compounds and thus overcome water deficit. The proportion of hexoses to sucrose, 1-kestose and nystose (SKN) was higher in WS plants. In WS 760, this increase was higher than in WS 380, and was not accompanied by decreases in SKN at the beginning of the treatment, as observed in WS 380 plants. These results suggest that the higher [CO2 ] in the atmosphere contributed to maintain, for a longer period, the pool of hexoses and of low DP fructans, favouring the maintenance of the water status and plant survival under drought.

Topics: Asteraceae; Brazil; Carbon Dioxide; Droughts; Fructans; Oligosaccharides; Photosynthesis; Soil; Sucrose; Trisaccharides; Water

The thin-layer chromatographic analysis of the crude oligosaccharides extracted from yacon revealed the presence of glucose, fructose, sucrose, 1-kestose and nystose. The qualitative and quantitative analysis was carried out on oligosaccharides by high pressure liquid chromatography and the results showed that the contents of d-glucose, fructose, sucrose, 1-kestose, nystose and 1-fructofuranosyl nystose in oligosaccharides were 38.30%, 16.44%, 14.58%, 12.29%, 12.17%, 6.20%, respectively. The content of the fructooligosaccharides in oligosaccharides was 30.66%. The crude oligosaccharides were separated and purified by silica gel column chromatography. The two fractions obtained from crude oligosaccharides were 1-kestose and nystose, which were identified by mass spectra. The yield of 1-kestose and nystose were 10.36% and 9.73%, respectively. The purity of 1-kestose was 82.9% and of nystose was 73.6%.

Topics: Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Oligosaccharides; Plant Extracts; Spectrometry, Mass, Electrospray Ionization; Trisaccharides

The utilization of 1-kestose (GF(2)) and nystose (GF(3)), the main components of fructooligosaccharides (FOS), by Lactobacillus and Bacteroides species was examined. Of seven Lactobacillus and five Bacteroides strains that utilized FOS, L. salivarius, L. rhamnosus, L. casei, and L. gasseri utilized only GF(2), whereas L. acidophilus and all the Bacteroides strains utilized both GF(2) and GF(3). Only the strains able to utilize both GF(2) and GF(3) had β-fructosidase activity in the culture supernatants. The culture supernatants of the Lactobacillus strains had higher β-fructosidase activity for GF(2) than for GF(3), whereas those of the Bacteroides strains had higher activity for GF(3) than for GF(2). Furthermore, β-fructosidase activity of the culture supernatants of the Lactobacillus cells grown in the GF(3) medium was much higher than that of the cells grown in the GF(2) medium, whereas the activity of the culture supernatants of the Bacteroides cells grown in the GF(3) medium was almost the same as that of the cells grown in the GF(2) medium. These results indicate that Lactobacillus species metabolize FOS in a different way from that of Bacteroides species.

Topics: Bacteroides; beta-Fructofuranosidase; Lactobacillus; Oligosaccharides; Species Specificity; Trisaccharides

Continuous production of fructooligosaccharides (FOS) by Aureobasidium pullulans immobilized on calcium alginate beads with a packed bed was investigated at a plant scale reactor. Optimum conditions were with 770 g sucrose/l, being fed at 200 l/h at 50°C which gave a productivity of 180 g FOS/l h. Initial activity was maintained for more than 100 days. The reactor was successfully scaled up to a production scale of 1.2 m(3).

Topics: Ascomycota; Bioreactors; Cells, Immobilized; Enzyme Stability; Glucose; Hexosyltransferases; Industrial Microbiology; Oligosaccharides; Sucrose; Trisaccharides

An extracellular beta-fructofuranosidase from the yeast Xanthophyllomyces dendrorhous was characterized biochemically, molecularly, and phylogenetically. This enzyme is a glycoprotein with an estimated molecular mass of 160 kDa, of which the N-linked carbohydrate accounts for 60% of the total mass. It displays optimum activity at pH 5.0 to 6.5, and its thermophilicity (with maximum activity at 65 to 70 degrees C) and thermostability (with a T(50) in the range 66 to 71 degrees C) is higher than that exhibited by most yeast invertases. The enzyme was able to hydrolyze fructosyl-beta-(2-->1)-linked carbohydrates such as sucrose, 1-kestose, or nystose, although its catalytic efficiency, defined by the k(cat)/K(m) ratio, indicates that it hydrolyzes sucrose approximately 4.2 times more efficiently than 1-kestose. Unlike other microbial beta-fructofuranosidases, the enzyme from X. dendrorhous produces neokestose as the main transglycosylation product, a potentially novel bifidogenic trisaccharide. Using a 41% (wt/vol) sucrose solution, the maximum fructooligosaccharide concentration reached was 65.9 g liter(-1). In addition, we isolated and sequenced the X. dendrorhous beta-fructofuranosidase gene (Xd-INV), showing that it encodes a putative mature polypeptide of 595 amino acids and that it shares significant identity with other fungal, yeast, and plant beta-fructofuranosidases, all members of family 32 of the glycosyl-hydrolases. We demonstrate that the Xd-INV could functionally complement the suc2 mutation of Saccharomyces cerevisiae and, finally, a structural model of the new enzyme based on the homologous invertase from Arabidopsis thaliana has also been obtained.

Topics: Basidiomycota; beta-Fructofuranosidase; DNA, Fungal; Enzyme Stability; Fungal Proteins; Glycoproteins; Hydrogen-Ion Concentration; Molecular Sequence Data; Molecular Weight; Oligosaccharides; Phylogeny; Protein Structure, Tertiary; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Substrate Specificity; Sucrose; Temperature; Trisaccharides

Fusarium head blight caused by Fusarium graminearum is a disease of cereal crops that not only reduces crop yield and quality but also results in contamination with trichothecenes such as nivalenol and deoxynivalenol (DON). To analyze the trichothecene induction mechanism, effects of 12 carbon sources on the production of DON and 3-acetyldexynivalenol (3ADON) were examined in liquid cultures incubated with nine strains of 3ADON-producing F. graminearum. Significantly high levels of trichothecene (DON and 3ADON) production by sucrose, 1-kestose and nystose were commonly observed among all of the strains tested. On the other hand, the levels of trichothecene biosynthesis induced by the other carbon sources were strain-specific. Tri4 and Tri5 expressions were up-regulated in the sucrose-containing medium but not in glucose. Trichothecene accumulation in the sucrose-containing medium was not repressed by the addition of glucose, indicating that trichothecene production was not regulated by carbon catabolite repression. These findings suggest that F. graminearum recognizes sucrose molecules, activates Tri gene expression and induces trichothecene biosynthesis.

Topics: Carbon; Fusarium; Gene Expression Profiling; Gene Expression Regulation, Fungal; Molecular Sequence Data; Oligosaccharides; Sucrose; Trichothecenes; Trisaccharides

A methods for the simultaneous determination for 1-kestose (GF2) and nystose (GF3) in foods and vegetables by high performance anion exchange chromatography with integrated pulsed amperometric detection (HPAEC-PAD) was developed in this study.. With a gradient program, and NaOH and sodium acetate as mobile phase, the two fructo-oligosaccharides (FOS) were separated on a CarboPac PA1 column and then detected by a gold electrode.. The linear range were 1.22 - 15.20 microg/g and 1.64 - 18.30 microg/g for GF2 and GF3, and the calculated method detection limit (MDL) were 0.085 microg/g and 0.089 microg/g, respectively. The relative standard deviation of GF2 (6.08 microg/g) and GF3 (7.32 microg/g) were 2.80% and 1.68% (n = 6). The contents of the two fructo- oligosaccharides in chicory root, artichoke and oral liquid were also determined. The results shows chicory root possess the highest GF2 and GF3 contents in all vegetable tissues, 6.61g/100g dw and 6.51g/100g dw, whereas the highest composition of kestose and nystose in food was found in Quantum FOS powder (raw material for additives), 34.13g/100g and 32.12g/100g respectively. The method recoveries of GF2 and GF3 were 93.97% and 97.14%.. This method was sensitive, selective, and simple for FOS quantification in vegetables and food products.

Topics: Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Electrochemistry; Food Analysis; Oligosaccharides; Trisaccharides; Vegetables

We have previously shown that non-digestible saccharides (NDS) stimulate intestinal Ca absorption via tight junctions. However, the cellular mechanisms activated by the NDS are not yet known. We investigated the effects of four NDS, difructose anhydride (DFA) III, DFAIV, fructo-oligosaccharides, and maltitol, on intracellular Ca signalling in isolated rat small-intestinal enterocytes. The changes in intracellular Ca(2+) concentration were measured before and after the addition of capric acid (7.5 or 15 mmol/l, a positive control), glycerol, or each NDS (1 or 10 mmol/l) to fura-2-loaded enterocytes. Treatment with capric acid or each NDS caused an immediate and dose-dependent rise in intracellular Ca(2+) concentration. Mechanical and osmotic stimulation achieved by adding glycerol had no effect on intracellular Ca(2+) concentration. The intracellular Ca(2+) concentration in enterocytes treated with DFAIII and fructo-oligosaccharides reached a peak level at about 30 s after stimulation, but those treated with DFAIV and maltitol showed further increases after the initial rapid rise. The maximum change in intracellular Ca(2+) concentration obtained by the application of maltitol was higher than that of DFAIII at 10 mmol/l. These findings suggest that each of the four NDS directly stimulates rat enterocytes, and increases intracellular Ca(2+) concentration. Thus, molecular structure may be more important than the size of the NDS in the induction of Ca signalling in the cells.

Topics: Animals; Antifungal Agents; Calcium; Decanoic Acids; Disaccharides; Enterocytes; Intestine, Small; Male; Maltose; Oligosaccharides; Rats; Rats, Sprague-Dawley; Sugar Alcohols; Sweetening Agents; Trisaccharides

By screening a cDNA library of artichoke (Cynara scolymus) blossom discs for fructosyltransferases, we isolated a clone designated Cy21. The deduced amino acid sequence shows homology to acid beta-fructosyl hydrolases and to the sucrose-fructan 6-fructosyltransferase (6-SFT) of barley. Transiently expressed in Nicotiana tabacum protoplasts, the Cy21 gene-product synthesized 1-kestose, indicating that Cy21 codes for a sucrose sucrose 1-fructosyltransferase (1-SST). The enzyme worked at physiologically relevant sucrose concentrations (25 mM sucrose). In the protoplast system, 1-kestose seemed to be the only fructan product of the 1-SST. The enzyme activity was not affected by pyridoxal-HCl, an inhibitor of both the beta-fructosyl hydrolase and the fructosyltransferase activity of invertases. The fructosyltransferase activity of the Cy21 gene-product, however, could be inhibited by Zn2+, Ag+ and Cu2+ ions. In artichoke plants the Cy21 transcript was highly abundant in primary roots and blossom discs. Transgenic potato tubers expressing Cy21 contain high levels of 1-kestose along with nystose and traces of fructosyl-nystose, supporting the conclusion that the Cy21 clone encodes a sucrose sucrose 1-fructosyltransferase.

Topics: Amino Acid Sequence; Cations, Divalent; Cloning, Molecular; Enzyme Inhibitors; Gene Library; Hexosyltransferases; Kinetics; Molecular Sequence Data; Nicotiana; Oligosaccharides; Plant Roots; Plants, Genetically Modified; Plants, Toxic; Protoplasts; Recombinant Proteins; Solanum tuberosum; Sucrose; Transfection; Trisaccharides; Vegetables

We developed a new type of anal cup for prevention of coprophagy and determined whether the absorption of Ca and Mg and the stimulatory effects of feeding fructo-oligosaccharides (FO) on the absorption of Ca and Mg were altered by prevention of coprophagy in rats. Rats were fed on a FO-free diet or a diet containing 50 g FO/kg for 2 weeks with or without prevention of coprophagy. FO-feeding increased the apparent absorptive ratio of Ca and Mg in rats with or without prevention of coprophagy. However, in the FO-fed groups the absorptive ratio of Mg in rats with prevention of coprophagy was higher than in rats without prevention of coprophagy. The Ca content of the femur was higher in rats fed on the FO-diet than in rats fed on the FO-free diet both with and without coprophagy. In conclusion, FO-feeding increased the absorption of Ca and Mg in rats both with and without coprophagy. Moreover, prevention of coprophagy enhanced the absorption of Mg in rats fed with FO. Coprophagy has to be considered when the effects of luminal fermentation or mineral absorption are examined in rats.

Topics: Animals; Calcium; Coprophagia; Fatty Acids, Volatile; Feces; Femur; Hydrogen-Ion Concentration; Intestinal Absorption; Magnesium; Male; Oligosaccharides; Rats; Rats, Sprague-Dawley; Trisaccharides

Topics: Carbohydrate Conformation; Carbohydrate Sequence; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Oligosaccharides; Sucrose; Trisaccharides