Enzymatic synthesis of fructooligosaccharides by inulinases from Aspergillus niger and Kluyveromyces marxianus NRRL Y-7571 in aqueous–organic medium

This work is focused on the synthesis of the fructooligosaccharides (FOS) from sucrose and inulin, using free, immobilized and pre-treated immobilized inulinase from Kluyveromyces marxianus NRRL Y 7571 and Aspergillus niger in an aqueous–organic system. Initially, the influence of pre-treatment using four different gases, propane, n-butane, CO₂ and liquefied petroleum gas (LPG), was investigated towards FOS production and best results were found when both enzymes were previously treated with LPG. The best reaction yields were obtained when the immobilized enzymes were treated with LPG. Considering FOS synthesis using the enzyme from A. niger, yields of 26.62% of GF2 (kestose), 30.62% of GF3 (nystose) and 8.47% of GF4 (fructosyl nystose) were achieved using sucrose as substrate. Using inulinases from K. marxianus NRRL Y 7571, 11.89% of GF2 and 20.83% of GF3 were obtained, using inulin as substrate. However, promising results were achieved using the free form of inulinase from A. niger (77.19% of GF2; 14.03% of GF3 and 0.07% of GF4) using inulin as substrate.     

Nutritional and functional characterisation of Andean chicuru (Stangea rhizanta)

A thorough functional and nutritional characterisation of Andean chicuru (Stangean rhizanta) root demonstrated its potential as an alternative source of fructooligosacharides (FOS), phenolic compounds, natural antioxidants and minerals. FOS of the GFn type with a degree of polymerisation (DP) of 3 (1-kestose) and 4 (nystose) were present. Important phenolic content and antioxidant activity values were obtained. The main phenolic compounds revealed by HPLC-DAD were caffeic and chlorogenic acids and their respective derivatives as well as flavan-3-ol derivatives. The nutritional analysis revealed high calcium and iron values, as well as considerable amounts of soluble and insoluble dietary fibre. The presence of low DP FOS, together with the high calcium and iron contents in chicuru, might favour its use for industrial applications.     

Fructo-oligosaccharides in table grapes and response to storage

Fructo-oligosaccharides (FOS) have been recognized as health food ingredients with a protective effect against environmental stresses in plants. We have analyzed the profiles of individual FOS in Cardinal table grape pulp, until now undetected, and quantified their changes in response to low temperature and high CO₂ levels. FOS separation and quantification was carried out using anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), and the glucose, fructose and sucrose content of the grapes was also determined. Five FOS were identified and quantified: 1-kestose, neokestose, nystose, nystose b and kestopentaose. While in non-treated table grapes the endogenous FOS remained at steady state levels during storage at 0 °C, exposure to 20% CO₂ for 3 days significant increases the levels of 1-kestose and kestopentaose, members of the inulin series. Considering the competitive advantage afforded by CO₂-treated grapes, this transitory FOS accumulation could provide protection against damage caused by low temperature storage.     

Recent trends in the microbial production,analysis and application of Fructooligosaccharides

This review focuses on the recent developments in the area of FOS research—its microbial production, functional properties and applications and an overview of the different analytical methods for the determination of the FTase. Different microbial sources of FTase reported in literature to produce FOS with different linkages to form 1-kestose, 6-kestose and neokestose in varying yields based on initial sucrose concentration is discussed. Different fermentative methods have been used for production of FOS. SSF has been used for the production of a value added product FOS utilizing various agroindustrial byproducts. The nutritional and culture parameters when optimized, the FOS yields and productivity could be improved. The use of immobilized enzymes and cells has led to the development of effective and economic methods for large-scale production of FOS. Forced flow Membrane reactor systems, biocatalyst system with a bioreactor equipped with a microfiltration systems, have been used for production of high content FOS by removing the released glucose and unreacted sucrose from the reaction mixture resulting in up to 98% FOS. The use of mixed enzyme system of Fructosyl Transferase and glucose oxidase or glucose dehydrogenase, could produce highly concentrated FOS up to 90–98%. Nano-filtration for removing glucose resulted in FOS of 90% concentration. The purified enzyme was found to produce kestose and nystose unlike the crude enzyme which produced GF₅ and GF₆ oligosaccharides Kinetic parameters (V_m, K_m, and K_i) of the enzyme were determined from experimental data on the transfructosylation rate at various substrate concentrations with and without addition of glucose Techniques like HPLC, using polar-bonded phase and resin-based HPLC columns are commonly used for separation of oligosaccharides with refractive Index Detector or pulsed amperometric detector and annular size exclusion chromatography for large scale and continuous fractionation. Other techniques like gas liquid chromatography, thin layer chromatography, NMR and Mass Spectrometry have been used for structure analyses. The functional properties like use as prebiotics, dietary fiber, role in absorption and defense/Immunity, lipid metabolism control of diabetics have been discussed. A variety of applications in food formulations are also discussed.     

Study of total fructan and fructooligosaccharide content in different onion tissues

The objective of this work was to determine the fructan and fructooligosaccharide (FOS) content of different onion tissues in order to evaluate the potential use of onion by‐products from the food industry as a source of FOS and fructans. Assays with two methods were carried out to optimise the extraction procedure. The main FOS, namely kestose (GF₂), nystose (GF₃) and fructofuranosylnystose (GF₄), were measured directly using standard sugars. The method for total fructans was based on enzymatic treatment (Novozym 230) of ethanolic/aqueous extract followed by determination of released fructose and glucose by HPLC. Data showed a clear predominance of GF₂ in every onion tissue and no occurrence of highly polymerised fructans. The tissues richest in fructans were the fleshy layers, so that the outer two fleshy layers turn out to be the best onion by‐product as a possible fructan source. © 2000 Society of Chemical Industry     

Physicochemical Properties of Bread Baked from Flour Blended with Immature Wheat Meal Rich in Fructooligosaccharides

ABSTRACT: Grain of the soft white wheat cultivar Harus was harvested weekly from anthesis to maturity and fructooligosaccharides (FOS) contents were determined by reversed-phase high-performance liquid chromatography. Tests were carried out to determine the effect of adding immature wheat meal to a base flour of cultivar Russ (hard red spring) on the quality characteristics of bread. FOS content was also analyzed in baked bread, and the effect of transglutaminase in improving bread quality was examined. Marked decreases in FOS contents, such as 1-kestose and nystose, were observed with grain maturation. The overall quality of bread appeared to be acceptable, and the added FOS were retained after baking.     

Fructo-oligosaccharides: Production,Purification and Potential Applications

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.     

高效离子色谱法分离、检测低聚果糖

采用CarboPac PA20柱和脉冲安培检测器的高效阴离子交换色谱建立一种低聚果糖的分析方法,该法可有效分离蔗果型和果果型低聚糖组分,并由此推测出低聚果糖的来源。结果表明,在优化条件下,蔗果低聚糖中蔗果三糖、新蔗果三塘、蔗果四糖和蔗果五糖的回收率分别为98.27%～99.72%、97.32%～102.56%、99.33%～101.81%和97.38%～102.12%。该法的相对标准偏差在0.55%～2.15%。蔗果三糖、新蔗果三塘、蔗果四糖和蔗果五糖的方法检出限分别为0.13、0.19、0.20 μg/g和0.41 μg/g。蔗果三糖在0.3～18 μg/mL、新蔗果三糖和蔗果四糖在0.5～20 μg/mL、蔗果五糖在0.8～23 μg/mL的范围内,其质量浓度与峰面积线性关系良好,相关系数在0.999 6～0.999 9之间。本方法简单、易于掌握,测定结果准确,适合低聚果糖产品及蛋白粉和奶粉等功能性食品中低聚果糖成分的检测分析。     

Qualitative and quantitative composition of fructooligosaccharides in bread

Short-chain fructooligosaccharides (FOS) of DP 3–5 in various bread were determined by means of HPLC method with the use of water–alcohol extracts. Crisp bread produced without yeast contains kestose, nystose and fructosyl-nystose in the total amount varying from 0.3 to 0.81 g FOS/100 g. Contents of short-chain fructooligosaccharides (FOS) of DP 3–5 in another kinds of bread examined–wheat, rye and wholemeal bread produced with the use of yeast–did not exceed 0.03 g/100 g DM.     

Lipolysis in probiotic and synbiotic cheese: The influence of probiotic bacteria, prebiotic compounds and ripening time on free fatty acid profiles

The influence of probiotic bacteria (Lactobacillus casei-01, Bifidobacterium lactis B94), prebiotic compounds (FOS and inulin) and ripening time (0-60 days) on the free fatty acid (FFA) profile of cheese, with special emphasis on the conjugated linoleic acid (CLA) content, was investigated. After 60 days of ripening, 10⁹-10¹⁰ cfu g⁻¹ cheese were recorded in both probiotic and synbiotic cheeses, despite harsh conditions of low pH values (4.1-5.1) and low moisture content (<30%, w/w). Increases in total FFA and CLA were observed throughout the ripening period, especially in synbiotic cheeses containing FOS and inulin (50:50) inoculated with B. lactis B94. The addition of FOS alone or combined with inulin did not significantly affect probiotic strain growth and viability during the ripening period; however, the advantage of the addition of prebiotic compounds in probiotic cheese manufacture is that it may allow the production of cheeses with improved performance as far as functional CLA compounds are concerned, as well as an improved nutritional quality reflected in a lower atherogenicity index.     

A nondigestible saccharide,fructooligosaccharide, increases the promotive effect of a flavonoid, α‐glucosyl‐isoquercitrin,on glucagon‐like peptide 1 (GLP‐1) secretion in rat intestine and enteroendocrine cells

This study conducted in vivo and in situ experiments with rats to investigate the glucagon‐like peptide‐1 (GLP‐1) secretion in response to oral or ileal administration of α‐glucosyl‐isoquercitrin (20–40 mmol in 2 mL; Q3G), fructooligosaccharides (200 mmol in 2 mL; FOS) and Q3G + FOS. Direct effects on GLP‐1‐producing l ‐cells were also examined by an in vitro study using a murine enteroendocrine cell line, GLUTag. To evaluate the plasma GLP‐1 level, blood samples from jugular cannula for in vivo and portal cannula for in situ experiments were obtained before and after administration of Q3G, FOS, or Q3G + FOS. We found tendencies for increases but transient stimulation of GLP‐1 secretion by Q3G in in vivo and in situ experiments. Although FOS alone did not have any effects, Q3G + FOS enhanced and prolonged high plasma GLP‐1 level in both experiments. In addition, application of Q3G on GLUTag cells stimulated GLP‐1 secretion while FOS enhanced the effect of Q3G. Our results suggest that Q3G + FOS possess the potential for the management or prevention of Type 2 diabetes mellitus (T2DM) by enhancing and prolonging the GLP‐1 secretion via direct stimulation of GLP‐1 producing l ‐cell.     

Anti-diabetic effect of the soybean extract fermented by Bacillus subtilis MORI in db/db mice

This study investigated the effects of soy bean extract fermented by Bacillus subtilis MORI (BTD-1) on blood glucose, hemoglobin A1c (HbA1c), plasma insulin, and pancreatic β islets in db/db mice. The BTD-1 (500 mg/kg) group showed significantly lower fasting blood glucose level (p<0.01) and postprandial 2 h blood glucose level (p<0.01) compared with the db control group. The BTD-1 (500 mg/kg) group showed significantly lower HbA1c level compared with the db control group (p<0.01). After 8 weeks of BTD-1 administration, the pancreatic islet architecture was preserved and the immunofluorescent intensities of insulin in BTD-1 (500mg/kg) group apparently increased compared to in the db control group. Plasma insulin levels were found to be significantly higher in the BTD-1 (500 mg/kg) group than in the db control group (p<0.05). In summary, our results suggest that BTD-1 has an anti-diabetes effect in a non-insulin dependent diabetes mellitus mouse model.     

Role of mono- and oligosaccharides from FOS as stabilizing agents during freeze-drying and storage of Lactobacillus delbrueckii subsp. bulgaricus

The aim of this work was to assess the role of mono- and oligosaccharides present in fructo-oligosaccharides (FOS) mixtures as protective agents during freeze-drying and storage of Lactobacillus delbrueckii subsp. bulgaricus CIDCA 333.Different FOS mixtures were enzymatically obtained from sucrose and further purified by removing the monosaccharides produced as secondary products. Their glass transition temperatures (T_g) were determined at 11, 22 and 33% relative humidity (RH). Bacterial cultures were freeze-dried in the presence of 20% w/v solutions of the studied FOS. Their protective effect during freeze-drying was assessed by bacterial plate counting, and by determining the lag time from growth kinetics and the uptake of propidium iodide (PI). Plate counting during bacterial storage at 4 °C, and 11, 22 and 33% RH for 80 days completed this rational analysis of the protective effect of FOS.Purification of FOS led to an increase of T_g in all the conditions assayed. Microorganisms freeze-dried in the presence of non-purified FOS were those with the shortest lag times. Bacteria freeze-dried with pure or commercial FOS (92% of total FOS) showed larger lag times (8.9–12.6 h). The cultivability of microorganisms freeze-dried with non-purified FOS and with sucrose was not significantly different from that of bacteria before freeze-drying (8.74 ± 0.14 log CFU/mL). Pure or commercial FOS were less efficient in protecting bacteria during freeze-drying. All the protectants prevented membrane damage. The cultivability of bacteria freeze-dried with FOS decayed < 1 logarithmic unit after 80 days of storage at 11% RH. When storing at 22 and 33% RH, pure and commercial FOS were those that best protected bacteria, and FOS containing monosaccharides were less efficient.The effect of FOS on bacterial protection is the result of a balance between monosaccharides, sucrose and larger FOS in the mixtures: the smallest sugars are more efficient in protecting lipid membranes, and the larger ones favor the formation of vitreous states.     

Production of fructooligosaccharides and β-fructofuranosidase by batch and repeated batch fermentation with immobilized cells of Penicillium expansum

The production of fructooligosaccharides (FOS) and ??-fructofuranosidase (FFase) by immobilized cells of Penicillium expansum was evaluated. In an initial stage, different low-cost materials including synthetic fiber, polyurethane foam, stainless steel sponge, loofah sponge, and cork oak were tested as carrier for the fungus immobilization. Additionally, the influence of the inoculum age (1 or 3?weeks) on cells immobilization, FOS and FFase production was also verified. Synthetic fiber and polyurethane foam were the best materials for P. expansum immobilization (2.21 and 1.98?g/g carrier, respectively) and FOS production (120.3 and 104.8?g/l), and gave also high results of FFase activity (23.01 and 32.42?U/ml). Then, the production of FOS and FFase by repeated batch fermentation with P. expansum immobilized on synthetic fiber was studied, aiming to improve the batch fermentation results. The results obtained in this stage were very promising with FOS yields of 87, 72, and 44?%, in the 3 initial cycles (60?h), respectively; the FFase activity was constant throughout the process (6 cycles, 96?h). Repeated batch fermentation with immobilized cells of P. expansum was found as being a technology with great potential for FOS and FFase production on industrial scale.     

Impact of Bifidobacterium animalis subsp. lactis BB-12 and, Lactobacillus acidophilus LA-5-containing yoghurt, on fecal bacterial counts of healthy adults

This randomized, placebo-controlled, double blind, parallel dose-response study investigated the impact of 4-week commercial yoghurt consumption supplemented with Bifidobacterium animalis subsp. lactis (BB-12) and Lactobacillus acidophilus (LA-5) on fecal bacterial counts of healthy adults. Fifty-eight volunteers were randomly assigned to three different groups: 1. placebo (no probiotic, no starter and no green tea extract); 2. Yoptimal (10⁹ cfu/100 g of BB-12 and LA-5 and 40 mg of green tea extract) and 3. Yoptimal-10 (10¹⁰ cfu/100 g of BB-12, 10⁹ cfu/100 g of LA-5 and 40 mg of green tea extract). These yoghurt products also contained Lactobacillus delbrueckii subsp. bulgaricus (10⁷ cfu/100 g) and Streptococcus thermophilus (10¹⁰ cfu/100 g). The quantitative PCR (qPCR) results showed that there were significant increases (P = 0.02) in bifidobacteria counts with the Yoptimal treatment as compared to baseline. The fecal numbers of B. animalis subsp. lactis and LA-5 significantly increased in the two probiotic treatments compared to the placebo treatment. Viable counts of fecal lactobacilli were significantly higher (P = 0.05) and those of enterococci were significantly lower (P = 0.04) after the intervention when compared to placebo. No significant difference was observed between treatments in volunteers' weight, waist girth, blood pressure, fasting plasma triglyceride and HDL-C concentrations, as well as cholesterol/HDL-cholesterol ratio. However, a significant increase in plasma cholesterol levels was observed in the placebo group (P = 0.0018) but the levels remained stable in the two probiotic yoghurt groups. These results show that probiotic strains supplemented in the form of yoghurt remain active during gut transit and are associated with an increase in beneficial bacteria and a reduction in potentially pathogenic bacteria. This trial was registered at clinicaltrials.gov as NCT00730626.     

Strawberry consumption improves plasma antioxidant status and erythrocyte resistance to oxidative haemolysis in humans

Significant increases in the plasma total antioxidant capacity (TAC) have already been reported after acute intake of strawberries. In addition, antihaemolitic effects of strawberry extracts have been recently demonstrated in vitro, revealing that part of the antioxidant properties of strawberry bioactive compounds could lie in their localisation within cell membranes. However, there is a lack of research evidence from in vivo protracted strawberry consumption studies. We carried out a 16-day pilot study where 12 healthy subjects ingested 500 g of antioxidants-rich strawberries daily, and we evaluated the potential effects of fruit consumption on biomarkers of plasma and cellular antioxidant status. A significant increase in fasting plasma TAC and in serum vitamin C concentrations were progressively observed during the period of strawberry supplementation. An enhanced resistance to haemolysis was also observed in both AAPH-treated and untreated erythrocytes, collected during and after the period of strawberry consumption. The results obtained in this work suggest that regular consumption of antioxidant-rich strawberries may exert an improvement on the plasma antioxidant status and an increase on the antihaemolitic defenses of human erythrocytes.     

Biochemical changes to milk following treatment by a novel,cold atmospheric plasma system

It has previously been shown that non-thermal (i.e., cold) plasma can successfully decontaminate milk from Escherichia coli. This study investigated the possible biochemical changes to the protein, free fatty acids and volatiles profiles of whole raw milk samples following application of cold plasma. Raw milk was treated with a cold plasma system at intervals of 0, 3, 6, 9, 12, 15 and 20 min. Significant changes were observed for 1 octanol (P < 0.05), 2 heptanone (P < 0.01), 2 hexenal (P < 0.01), 2 octenal (P < 0.05), nonanal and benzaldehyde (P < 0.001). Plasma treatment did not result in significant changes to the lipid composition of raw milk. However, exposure to cold plasma significantly increased the total aldehyde content following 20 min treatment. No significant difference was observed in the total ketone or alcohol levels.     

Competition mechanisms of lactic acid bacteria and bifidobacteria: Fermentative metabolism and colonization

Enzyme activities (α- and β-glucosidases, α- and β-galactosidases and β-fructofuranosidase) and organic acid production of four strains of lactic acid bacteria (LAB; Streptococcus thermophilus STY-31, Lactobacillus delbrueckii subsp. bulgaricus LBY-27, Lactobacillus casei LC-01 and Lactobacillus acidophilus LA-5) and Bifidobacterium lactis BB-12 were tested on milk and MRS fermentation broth with glucose, lactose or fructooligosaccharides (FOS) as carbon source. The highest β-galactosidase activity was found in L. acidophilus growing on milk. As compared to milk, α-glucosidase activity was increased with FOS by B. lactis, L. acidophilus and L. casei. The analysis of organic acids and short-chain fatty acids in the medium growth showed that lactate and acetate were the major fermentation metabolites produced by LAB and bifidobacteria, respectively. However, a metabolic shift towards more acetate and formate production, at the expense of lactate production, was observed during growth of L. casei on FOS. When grown on FOS as sole carbon source, L. acidophilus showed the highest production of lactate among the species tested. In addition, L. acidophilus demonstrated resistance to colonization against the intestinal pathogens Escherichia coli and Salmonella enterica in competition assays.