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Levan is a bioactive polysaccharide that can be synthesized by various microorganisms. In this study, the physicochemical properties and bioactivity of levan synthesized by recombinant levansucrase from Erwinia tasmaniensis were investigated. The synthesis conditions, including the enzyme concentration, substrate concentration, and temperature, were optimized. The obtained levan generally appeared as a cloudy suspension. However, it could transform into a hydrogel at concentrations exceeding 10 % (w/v). Then, ultrasonication was utilized to reduce the molecular weight and increase the bioavailability of levan. Dynamic light scattering (DLS) and gel permeation chromatography (GPC) indicated that the size of levan was significantly decreased by ultrasonication, whereas Fourier transform infrared spectroscopy

Plastic pollution is increasing day by day and the search for new, environmentally friendly products continues. Herein, for the first time, different degrees of mucilage were obtained from chia seeds and the film-forming behavior of levan biopolymer with these mucilages was investigated. Glycerol and sorbitol were used as plasticizers in the film design. Films prepared with sorbitol were characterized physically, mechanically and morphologically. The antioxidant and antimicrobial effects of the films were examined. The films formed as nanocomposites of levan and chia seed mucilages obtained at different temperatures (25 °C, 55 °C and 80 °C) exhibited structurally and mechanically different properties. It was observed that the films obtained with chia mucilages and levan preserved their antibacterial properties but lost their antifungal properties. In addition, quorum sensing property of the mucilage obtained at 55 °C during the investigation of the antibacterial property was reported for the first time with this study. The levan-based chia seed mucilages films obtained have the potential to be used in industrial and medical fields, and the nature-friendly nature of these films is very important for our green world.

Erwinia amylovora, the causative agent of fire blight, uses flagella-based motilities to translocate to host plant natural openings; however, little is known about how this bacterium migrates systemically in the apoplast. Here, we reveal a novel surface motility mechanism, defined as sliding, in E. amylovora. Deletion of flagella assembly genes did not affect this movement, whereas deletion of biosynthesis genes for the exopolysaccharides (EPSs) amylovoran and levan resulted in non-sliding phenotypes. Since EPS production generates osmotic pressure that potentially powers sliding, we validated this mechanism by demonstrating that water potential positively contributes to sliding. In addition, no sliding was observed when the water potential of the surface was lower than -0.5 MPa. Sliding is a passive motility mechanism. We further show that the force of gravity plays a critical role in directing E. amylovora sliding on unconfined surfaces but has a negligible effect when cells are sliding in confined microcapillaries, in which EPS-dependent osmotic pressure acts as the main force. Although amylovoran and levan are both required for sliding, we demonstrate that they exhibit different roles in bacterial communities. In summary, our study provides fundamental knowledge for a better understanding of mechanisms that drive bacterial sliding motility.

Halomonas smyrnensis AAD6

Fructans are important biocompounds because of their health-promoting effects as dietary fiber and prebiotics and also because of their harmful effects as fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAP) particularly in people suffering from irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD), and recently as potential triggers of non-celiac wheat/gluten sensitivity. In this work, we have analyzed the fructan contents as well as its degree of polymerization (DP) in a genetically diverse set of wheat varieties, modern and landraces, from different commonly consumed species (

Three signal peptides from α-mating factor (α-MF), inulinase (INU) and native levansucrase (LS) were compared for secretion efficiency of Bacillus subtilis levansucrase SacB-T305A in Pichia pastoris GS115. The first complete secretion of bacterial levansucrase in yeasts under methanol induction was achieved while using α-MF signal. The secreted recombinant Lev(α-MF) proved to be glycosylated by combination of NanoLC-MS/MS and Endo H digestion. Interestingly, glycosylation not only improved significantly the polymerase thermostability, but also reversed the products profiles to favor synthesis of high molecular weight (HMW) levan which accounted for approximately 73 % to total levan-type polysaccharides. It indicated for the first time that the glycosylation of recombinant B. subtilis levansucrase affected significantly the products molecular weight distribution. It also provided a promising enzymatic way to effectively product HMW levan from sucrose resources.

The rhizosphere context of inulin-accumulating plants, such as Jerusalem artichoke (Helianthus tuberosus), is an ideal starting basis for the discovery of inulolytic enzymes with potential for bio fructose production. We isolated a Glutamicibacter mishrai NJAU-1 strain from this context, showing exo-inulinase activity, releasing fructose from fructans. The growth conditions (pH 9.0; 15 °C) were adjusted, and the production of inulinase by Glutamicibacter mishrai NJAU-1 increased by 90% (0.32 U/mL). Intriguingly, both levan and inulin, but not fructose and sucrose, induced the production of exo-inulinase activity. Two exo-inulinase genes (inu1 and inu2) were cloned and heterologously expressed in Pichia pastoris. While INU2 preferentially hydrolyzed longer inulins, the smallest fructan 1-kestose appeared as the preferred substrate for INU1, also efficiently degrading nystose and sucrose. Active site docking studies with GFn- and Fn-type small inulins (G is glucose, F is fructose, and n is the number of β (2-1) bound fructose moieties) revealed subtle substrate differences between INU1 and INU2. A possible explanation about substrate specificity and INU's protein structure is then suggested. KEY POINTS: • A Glutamicibacter mishrai strain harbored exo-inulinase activity. • Fructans induced the inulolytic activity in G. mishrai while the inulolytic activity was optimized at pH 9.0 and 15 °C. • Two exo-inulinases with differential substrate specificity were characterized.

A new exopolysaccharide (EPS) producing Gram-positive bacterium was isolated from the rhizosphere of Bouteloua dactyloides (buffalo grass) and its EPS product was structurally characterized. The isolate, designated as LB1-1A, was identified as Bacillus paralicheniformis based on 16S rRNA gene sequence and phylogenetic tree analysis. The EPS produced by LB1-1A was identified as a levan, having β(2 → 6) linked backbone with β(2 → 1) linkages at the branch points (4.66%). The isolate LB1-1A yielded large amount (~ 42 g/l) of levan having high weight average molecular weight (Mw) of 5.517 × 10

The influence of agave fructans (AF) (1-10%) and xanthan (from 0.03% to 0.25%) in combination with sodium caseinate (SC) at 1% on the rheological and physicochemical properties of aqueous phases and emulsions was evaluated. Steady-state flow behavior, particle size distribution, and stability studies were used to characterize the systems. The aqueous systems displayed the shear-thinning behavior characteristic of xanthan solutions; however, this behavior was modified by the presence of SC and AF due to interactions between AF-SC and AF-xanthan based on predominant hydrogen bonding because of the hydroxyl groups on AF. In emulsions, an increase in viscosity due to the effect of the AF concentration reflects a probable association of fructan aggregates on the surface of SC particles that reinforce the interfacial layer of SC, while xanthan contributes to an increase in the viscosity of the continuous phase, which effectively prevents coalescence and floc formation even at higher concentrations, despite the possible existence of a depletion flocculation effect attenuated by the interaction between AF-SC and AF-xanthan. PRACTICAL APPLICATION: These results can be of use, in an important way, in the design of stable functional emulsions in which there is an application for agave fructans recognized as dietary fiber, also considering their peculiar way of interacting with xanthan favoring its stabilizing functionality.

This symposium includes thirteen personal narratives from people who have received at least one organ transplant from a living or deceased donor. These narratives foster better understanding of the experiences of life-saving organ recipients and their families, including post-transplant difficulties experienced-sometimes requiring multiple transplants. This issue also includes three commentaries by Macey L. Levan, Heather Lannon, and Vidya Fleetwood, Roslyn B. Mannonamp; Krista L. Lentine. Dr. Levan is a living kidney donor and associate professor of surgery and population health. She writes and teaches on organ transplant and has expertise and experience in governance at the federal level as a member of the board of directors for the U.S. Organ Procurement and Transplantation Network. Dr. Lannon is a social worker and transplant advocate. Her work and interest in the topic were inspired by her husband who needed a heart transplant. Dr. Fleetwood is a surgeon specializing in liver transplantation, pancreas transplantation, kidney transplantation, as well as living donor nephrectomy. Dr. Mannon is a nephrologist and professor of medicine, pathology and microbiology. Her work focuses on understanding and improving long-term kidney transplant outcomes for patients, donors, and caregivers. Dr. Lentine is a nephrologist specializing in treatment of adult patients with kidney disease. She is a researcher and co-chaired the international work group that developed guidelines for living donor kidney transplantation.

Fructans such as inulin and levan accumulate in certain taxonomic groups of plants and are a reserve carbohydrate alternative to starch. Onion (Allium cepa L.) is a typical plant species that accumulates fructans, and it synthesizes inulin-type and inulin neoseries-type fructans in the bulb. Although genes for fructan biosynthesis in onion have been identified so far, no genes for fructan degradation had been found. In this study, phylogenetic analysis predicted that we isolated a putative vacuolar invertase gene (AcpVI1), but our functional analyses demonstrated that it encoded a fructan 1-exohydrolase (1-FEH) instead. Assessments of recombinant proteins and purified native protein showed that the protein had 1-FEH activity, hydrolyzing the β-(2,1)-fructosyl linkage in inulin-type fructans. Interestingly, AcpVI1 had an amino acid sequence close to those of vacuolar invertases and fructosyltransferases, unlike all other FEHs previously found in plants. We showed that AcpVI1 was localized in the vacuole, as are onion fructosyltransferases Ac1-SST and Ac6G-FFT. These results indicate that fructan-synthesizing and -degrading enzymes are both localized in the vacuole. In contrast to previously reported FEHs, our data suggest that onion 1-FEH evolved from a vacuolar invertase and not from a cell wall invertase. This demonstrates that classic phylogenetic analysis on its own is insufficient to discriminate between invertases and FEHs, highlighting the importance of functional markers in the nearby active site residues.

Inulin consumption in both humans and animal models is recognized for its prebiotic action with the most consistent change that lies in enhancing the growth and functionality of Bifidobacterium bacteria, as well as its effect on host gene expression and metabolism. Further, inulin-type fructans are utilized in the colon by bacterial fermentation to yield short-chain fatty acids (SCFAs), which play important role in its biological effects both locally inside the gut and in systemic actions. The gut symbiosis sustained by inulin supplementation among other dietary fibers exerts preventive and/or therapeutic options for many metabolic disorders including obesity, type 2 diabetes mellitus, cardiometabolic diseases, kidney diseases and hyperuricemia. Although, gastrointestinal negative effects due to inulin consumption were reported, such as gastrointestinal symptoms in humans and exacerbated inflammatory bowel disease (IBD) in mice. This comprehensive review aims to present the whole story of how inulin functions as a prebiotic at cellular levels and the interplay between physiological, functional and immunological responses inside the animal or human gut as influenced by inulin in diets, in context to its structural composition. Such review is of importance to identify management and feed strategies to optimize gut health, for instance, consumption of the tolerated doses to healthy adults of 10 g/day of native inulin or 5 g/day of naturally inulin-rich chicory extract. In addition, inulin-drug interactions should be further clarified particularly if used as a supplement for the treatment of degenerative diseases (e.g., diabetes) over a long period. The combined effect of probiotics and inulin appears more effective, and more research on this synergy is still needed.

There is great interest in the search for new alternatives to antimicrobial drugs, and the use of prebiotics and probiotics is a promising approach to this problem. This study aimed to assess the effect of inulin-type fructans, used in synbiotic combinations with Lactobacillus paracasei or Lactobacillus plantarum, on the production of short-chain fatty acids and antimicrobial activity against Candida albicans. The inhibition assay using the L. paracasei and L. plantarum supernatants resulting from the metabolization of inulin-type fructans displayed growth inhibition and antibiofilm formation against C. albicans. Inhibition occurred at concentrations of 12.5, 25, and 50% of the L. paracasei supernatant and at a concentration of 50% of the L. plantarum supernatant. The analysis of short-chain fatty acids by gas chromatography showed that lactic acid was the dominating produced metabolite. However, acetic, propionic, and butyric acids were also detected in supernatants from both probiotics. Therefore, the synbiotic formulation of L. paracasei or L. plantarum in the presence of inulin-type fructans constitutes with anticandidal effect is a possible option to produce antifungal drugs or antimicrobial compounds.

Carbohydrates such as fructans can be involved in priming or defence stimulation, and hence potentially provide new strategies for crop protection against biotic stress. Chicory (Cichorium intybus) is a model plant for fructan research and is a crop with many known health benefits. Using the chicory-Botrytis cinerea pathosystem, we tested the effectiveness of fructan-induced immunity, focussing on different plant and microbial fructans. Sugar dynamics were followed after priming and subsequent pathogen infection. Our results indicated that many higher plants might detect extracellular levan oligosaccharides (LOS) of microbial origin, while chicory also detects extracellular small inulin-type fructooligosaccharides (FOS) of endogenous origin, thus differing from the findings of previous fructan priming studies. No clear positive effects were observed for inulin or mixed-type fructans. An elicitor-specific burst of reactive oxygen species was observed for sulfated LOS, while FOS and LOS both behaved as genuine priming agents. In addition, a direct antifungal effect was observed for sulfated LOS. Intriguingly, LOS priming led to a temporary increase in apoplastic sugar concentrations, mainly glucose, which could trigger downstream responses. Total sugar and starch contents in total extracts of LOS-primed leaves were higher after leaf detachment, indicating they could maintain their metabolic activity. Our results indicate the importance of balancing intra- and extracellular sugar levels (osmotic balance) in the context of 'sweet immunity' pathways. ispartof: JOURNAL OF EXPERIMENTAL BOTANY vol:73 issue:12 pages:4214-4235 ispartof: location:England status: published

The present work reported the extraction, purification and characterization of an inulin fructan from Codonopsis pilosula (CPW1) and its application in stabilization of selenium nanoparticles (SeNPs). The morphology, stability, and stabilization mechanism of CPW1 stabilized SeNPs (CPW1-Se) were explored, and the results showed that the SeNPs were amorphous state, with size of 54-79 nm, and kept stable within 15 days due to the interaction between SeNPs and the hydroxyl groups on the surface of CPW1. Moreover, the effects on proliferation and apoptosis of CPW1-Se to both normal cells (293T) and liver cancer cells (Huh-7 and HepG2) were evaluated systematically by using the CCK8 assay, plate clone formation assay, flow cytometry and western blot. The results indicated that CPW1-Se possessed selective anti-hepatoma activities without side effects on normal cells, which exhibited strong potential application in liver cancer treatments.

This study aimed to evaluate the effect of supplementation with inulin-type fructans (ITFs) on the intestinal immune function in the context of dysbiosis resulting from antibiotic cocktail (ABx) treatment. BALB/c mice (8-9 weeks of age) were treated with an ABx for 3 weeks and then allowed to recover spontaneously or with ITF supplementation (5%) for 4 weeks. Our results showed that ABx treatment can induce gut microbiota dysbiosis and intestinal inflammation in mice. After 4 weeks of recovery, ITF supplementation restored the composition of the intestinal microbial community. However, compared with spontaneous recovery, ITF supplementation delayed inflammation recovery in the intestine and upregulated diamine oxidase (DAO) activity and increased lipopolysaccharide (LPS) content in serum. In addition, ITF supplementation delayed the regulatory T (Treg) cell and B cell recovery in the lamina propria (LP). Furthermore, compared with spontaneous recovery, ITF supplementation inhibited the relative expression of certain proinflammatory genes, such as for inducible nitric oxide synthase (iNOS) and tumour necrosis factor α (Tnf-α), in the colon, but it reduced the secretion of the anti-inflammatory mediator transforming growth factor β1 (TGF-β1) in serum, reduced the secretion of secretory immunoglobulin A (SIgA) in the colon and promoted the secretion of the proinflammatory cytokine interleukin (IL)-17A. In conclusion, these data supported the hypothesis that the influence of ITFs on the host's intestinal status is not always beneficial in the context of ABx-induced biological disorder. However, the significance of these findings needs to be determined by advanced studies KEY POINTS: • ITFs did not promote the recovery of microbial community composition. • ITFs delayed the recovery of ABx-induced colonic inflammation. • ITFs reduced the secretion of TGF-β1 and SIgA. • ITFs delayed the recovery of Treg and B cells in the LP.

Polycystic ovary syndrome (PCOS) is associated with several cardiovascular risk factors. Prebiotics were proposed to beneficially affect risk factors associated with metabolic disorders. The aim of this study was to investigate and compare the effects of inulin-type fructans (ITFs), as well-studied prebiotics, with different degrees of polymerization, on markers of inflammation, oxidative stress and endothelial dysfunction in PCOS patients.A randomized, double-blind, placebo-controlled trial.Seventy-five PCOS women were randomly assigned to receive 10 g/day of either high-performance inulin (HPI) or oligofructose-enriched inulin (OEI) or placebo for 12 weeks.Biochemical indices and blood pressure levelswere assessed before and after the intervention.In the intent-to-treat analysis, high-sensitive C-reactive protein (hs-CRP) decreased in HPI and OEI groups, over the 12 weeks, and the changes were significant in the HPI group, compared to placebo (changes from baseline in the HPI group: -0.11 vs. placebo group: 0.004 mg/L [conversion factor to SI units (nmol/L): 9/5238]; p = .007). Serum levels of nitric oxide (NO) increased, and endothelin-1 and total oxidant status decreased in HPI and OEI groups, at the end of the trial; however, these changes were not significantly compared to placebo (p = .07, .36 and .22, respectively). No differences in systolic and diastolic blood pressure were found. Per-protocol analysis (n = 68) yielded consistent results for all endpoints, with the exception that the significant effect of ITFs on serum hs-CRP levels in the unadjusted ITT analysis became nonsignificant in the per-protocol analysis (p = .06).A 12-week supplementation with long-chain ITFs had favourable effects on inflammatory status among PCOS patients.

FODMAPs produce similar small bowel water and colonic gas in patients with irritable bowel syndrome (IBS) and healthy controls (HCs), despite IBS patients reporting increased gastrointestinal (GI) symptoms.To unravel the mechanisms underlying FODMAP-induced symptom reporting, we investigated gut and brain responses to fructan administration in IBS patients and HC.This randomised, double-blind, cross-over study consisted of three visits where fructans (40 g/500 mL saline), glucose (40 g/500 mL saline) or saline (500 mL) were infused intragastrically during 1 h MR brain scanning; abdominal MRI was performed before, 1 h, and 2 h post-infusion. Symptoms were rated using validated scales.In IBS (n = 13), fructans induced more cramps, pain, flatulence and nausea compared to glucose (P = 0.03, 0.001, 0.009 and0.001 respectively), contrary to HC (n = 13) (all P 0.14), with between-group differences for cramps and nausea (P = 0.004 and 0.023). Fructans increased small bowel motility and ascending colonic gas and volume equally in IBS and HC (between-group P 0.25). The difference in colonic gas between fructans and saline covaried with differences in bloating and cramps in IBS (P = 0.008 and 0.035 respectively). Pain-related brain regions responded differentially to fructans in IBS compared to HC, including the cerebellum, supramarginal gyrus, anterior and midcingulate cortex, insula and thalamus (pFructans increase small bowel motility and colon gas and volume similarly in IBS patients and HC. Increased symptom responses to fructans in IBS covary with altered brain responses in pain-related regions, indicating that gut-brain axis dysregulation may drive FODMAP-induced symptom generation in IBS.

Here, gold nanoparticles (AuNPs) were synthesized upon exposure to nano-pulsed Nd-YAG laser irradiation in de-ionized water (PLAL) and functionalized with levan polysaccharide for assessing the anticancer efficacy of doxorubicin (DOX)-conjugated levan-capped AuNPs complexes to MCF-7 breast cancer cells. According to the physicochemical test results, the increments in levan amount enhanced the colloidal stability and the drug encapsulation efficiency (DEE) significantly. For the 10L-AuNP group having the highest levan amount (10 mg/mL levan), DEE was calculated as 92.21 ± 0.56%. The lean levan, uncapped AuNPs, and 10L-AuNP were found non-cytotoxic (80% cell viability) in the studied concentrations with 48 h MTT assays. At higher DOX loadings (25, 50, and 100 μg/mL) of 10L-AuNP, the cell viability reduced significantly compared to free DOX. Overall, these nanoparticle complexes could be proposed as potent drug delivery vehicles for cancer drugs such as DOX, as well as other drugs in the prospective studies.

Rye flour is used as the main ingredient of sourdough bread, which has technological and gastronomic benefits and increased nutritional value. The transformations observed during fermentation and baking may enable the conversion or degradation of rye dietary fiber carbohydrates built mainly of arabinoxylans, fructans, and β-glucans. This study aimed to determine the dynamics of the changes in the contents of complex carbohydrates in sourdoughs inoculated with potential probiotic microorganisms as well as the polysaccharide composition of the resulting bread. Sourdoughs were inoculated with the potential probiotic microorganisms Saccharomyces boulardii, Lactiplantibacillus plantarum, Lacticaseibacillus rhamnosus, and Bacillus coagulans, and spontaneous fermentation was performed as a control. Samples of the sourdoughs after 24 and 48 h of fermentation and of bread obtained with these sourdoughs were analyzed for the content of individual dietary fiber components. The present study demonstrated that the treatments applied contributed to an increased total content of arabinoxylans in the breads, and the inoculation of the sourdoughs with the potential probiotic strains improved their solubility in water. The use of the S.boulardii strain may seem prospective as it allowed for the greatest reduction in fructans in the rye bread. Rye sourdough bread is an attractive source of dietary fiber and can be modified for different nutritional needs.

FODMAPs are fermentable oligo-, di-, monosaccharides, and polyols. The application of homofermentative lactic acid bacteria (LAB) has been investigated as a promising approach for producing low-FODMAP whole-wheat bread. The low-FODMAP diet is recommended to treat irritable bowel syndrome (IBS). Wheat flour is staple to many diets and is a significant source of fructans, which are considered FODMAPs. The reduction of fructans via sourdough fermentation, generally associated with heterofermentative lactic acid bacteria (LAB), often leads to the accumulation of other FODMAPs. A collection of 244 wild-type LAB strains was isolated from different environments and their specific FODMAP utilisation profiles established. Three homofermentative strains were selected for production of whole-wheat sourdough bread. These were Lactiplantibacillus plantarum FST1.7 (FST1.7), Lacticaseibacillus paracasei R3 (R3), and Pediococcus pentosaceus RYE106 (RYE106). Carbohydrate levels in flour, sourdoughs (before and after 48 h fermentation), and resulting breads were analysed via HPAEC-PAD and compared with whole-wheat bread leavened with baker’s yeast. While strain R3 was the most efficient in FODMAP reduction, breads produced with all three test strains had FODMAP content below cut-off levels that would trigger IBS symptoms. Results of this study highlighted the potential of homofermentative LAB in producing low-FODMAP whole-wheat bread.

This review summarizes the nature, types, and properties of inulin polysaccharides and their applications as prebiotic dietary fibers. Natural food and commercial plant sources of inulin and extraction methods are presented. The physicochemical and functional properties of inulin are summarized. The prebiotic roles of inulin and their mechanisms of action are detailed. Inulin acts as prebiotic dietary fiber with multiple putative health benefits. It reduces caloric intake and contributes to reduced blood glucose and plasma lipid/cholesterol levels when used as sugar and fat replacers. It also stimulates immune systems and protects the colon mucosa against carcinogenesis and inflammation. Inulin also alters the composition and population of the gut microbiota. It stimulates the growth and activities of health beneficial microorganisms while inhibiting enteropathogenic bacteria. The beneficial microorganisms ferment inulin and produces acids including short‐chain fatty acids that lower the pH in the colon and inhibit pathogens. The health beneficial bacteria also produce other metabolites that positively influence human health. The consumption of inulin is however, associated to symptoms of gastrointestinal discomfort, when consumed at higher levels to meet the daily recommendation of dietary fiber. Potential solutions to the limitations are forwarded as future research ideas and policy inputs.

Gut microbial fermentation of soluble dietary fibers promotes general and substrate-specific health benefits. In this study, the fermentation characteristics of two soluble branched-dietary fibers, namely, agavin (a type of agave fructans) and digestion-resistant maltodextrin (RD) were investigated against cellulose, using a simulated colonic fermenter apparatus employing a mixed culture of swine fecal bacteria. After 48 h of complete fermentation period, the microbial composition was different among all groups, where Bifidobacterium spp. and Lactobacillus spp. dominated the agavin treatment, while the members of the families Lachnospiraceae and Prevotellaceae dominated the RD treatment. Agavin treatment exhibited a clearly segregated two-phased prolonged fermentation trend compared to RD treatment as manifested by the fermentation rates. Further, the highest short-chain fatty acids production even at the end of the fermentation cycle, acidic pH, and the negligible concentration of ammonia accumulation demonstrated favorable fermentation attributes of agavin compared to RD. Therefore, agavin might be an effective and desirable substrate for the colonic microbiota than RD with reference to the expressed microbial taxa and fermentation attributes. This study revealed a notable significance of the structural differences of fermentable fibers on the subsequent fermentation characteristics.

The complex of polysaccharides of the grain transforms during processing and modifies the physical and chemical characteristics of bread. The aim of the research was to characterize the changes of glucans, mannans and fructans in hull-less barley and wholegrain wheat breads fermented with spontaneous hull-less barley sourdough, germinated hull-less barley sourdough and yeast, as well as to analyze the impact of polysaccharides on the physical parameters of bread. By using the barley sourdoughs for wholegrain wheat bread dough fermentation, the specific volume and porosity was reduced; the hardness was not significantly increased, but the content of β-glucans was doubled. Principal component analysis indicates a higher content of β-glucans and a lower content of starch, total glucans, fructans and mannans for hull-less barley breads, but wholegrain wheat breads fermented with sourdoughs have a higher amount of starch, total glucans, fructans and mannans, and a lower content of β-glucans. The composition of polysaccharides was affected by the type of flour and fermentation method used.

Rising temperatures and pronounced drought are significantly affecting biodiversity worldwide and reducing yields and quality of Brassica crops. To elucidate the mechanisms of tolerance, 33 kale accessions (

Herbs and their combinations due to the wide range of biologically active substances can influence on various links of the pathogenetic mechanism of development of diabetes mellitus and its complications. One of such combinations is an antidiabetic herbal mixture with established hypoglycemic, hypolipidemic, antioxidant, hepatoprotective, pancreatoprotective activity in previous pharmacological study in vivo that including an inulin-containing component – Taraxacum officinale L. roots. Thus, the aim of this study was to determine the quantitative content of inulin and fructans in Taraxacum officinale L. Quantity content of inulin was determined by the difference between fructose as a product of enzymatic hydrolysis and D-fructose, a constituent of sucrose and free D-fructose, taking into account the empirical factor for the conversion of D-fructose from inulin. Carbohydrates used in the calculation of inulin were separated by gas chromatography-mass spectrometry after conversion into volatile derivatives as aldononitrile acetate. According to the results, Taraxacum officinale L. roots contain 436.29 mg/g of inulin. Total content of fructans was determined by spectrophotometric analysis as a product of acid hydrolysis of 5-(hydroxymethyl)furfural. The results show that Taraxacum officinale L. roots contain 39.49% of fructans. The obtained results are evidence that this plant component should be included in the herbal antidiabetic mixture, because due to the presence of fructans and inulin causes hypoglycemic, hypolipidemic and detoxification activity.

This article comments on: Oku S, Ueno K, Sawazaki Y, Maeda T, Jitsuyama Y, Suzuki T, Onodera S, Fujino K, Shimura H. 2022. Functional characterization and vacuolar localization of fructan exohydrolase derived from onion (Allium cepa). Journal of Experimental Botany 73,4908–4922. ispartof: JOURNAL OF EXPERIMENTAL BOTANY vol:73 issue:14 pages:4620-4623 ispartof: location:England status: published

Fructan is a polysaccharide composed of fructose and can be classified into several types, such as inulin, levan, and fructo-oligosaccharides, based on their linkage patterns and degree of polymerization. Owing to its structural and functional diversity, fructan has been used in various fields including prebiotics, foods and beverages, cosmetics, and pharmaceutical applications. With increasing interest in fructans, efficient and straightforward production methods have been explored. Since the 1990s, yeast cells have been employed as producers of recombinant enzymes for enzymatic conversion of fructans including fructosyltransferases derived from various microbes and plants. More recently, yeast cell factories are highlighted as efficient workhorses for fructan production by direct fermentation. In this review, recent advances and strategies for fructan biosynthesis by yeast cell factories are discussed.

Levan-type fructooligosaccharides (L-FOSs) are a prominent class of non-digestible oligosaccharides with potential as nutritional prebiotics. Endo-levanase, which randomly hydrolyzes β-(2,6)-linkages in fructans, is a promising enzyme for short-chain FOS production. In this work, a recombinant levanase (LevBk) from Bacillus koreensis strain HL12 was characterized. Soluble LevBk protein was produced in Escherichia coli BL21(DE3) system at 40 mg/L of culture medium. Based on sequence and structural analysis, LevBk was classified as a member of endo-levanase in GH32 family containing N-terminal substrate binding pocket and C-terminal β-sandwich domains. LevBk optimally worked at 45 °C, pH 6.0 with the specific activity of 2.43 U/mg. Based on enzymatic hydrolysis, short-chain L-FOSs with degree of polymerization (DP) of 2-4 were produced from hydrolysis of timothy grass levan under optimal conditions for 9-24 h. With its ability to produce L-FOSs with specific chain lengths, LevBk could be attractively applied for converting of levan containing material to high value-added sweetener in the biorefinery industry.

Despite the recognition of Agave tequilana Weber var. Azul as raw material for producing tequila and obtaining prebiotics, there are other highly relevant Agave species in Mexico. Oaxaca contains a startlingly diverse range of Agave species; Agave angustifolia Haw. and Agave potatorum Zucc. are two classic specimens with great commercial potential. In this study, we examined the fructan fluctuation in these two species during their lifetime in the field (from 1 to 6 years old). First, we analyzed their morphological diversity based on vegetative characteristics. Subsequently, fructan extracts were analyzed by TLC, FT-IR, and HPAEC-PAD to identify carbohydrates. Multivariate analyses of the morphological parameters indicated a morphological divergence between the two species. Furthermore, we found that the concentration of simple carbohydrates and fructans, as well as the fructan DP, changed during plant development. Glucose, fructose, and fructooligosaccharides (FOS) were more abundant in A. potatorum, while A. angustifolia showed a greater amount of sucrose and fructans with a high DP. Fructan DP heatmaps were constructed using HPAEC-PAD profiles—the heatmaps were very helpful for establishing an easy correlation between age and the carbohydrate types present in the fructan extracts. This study is an important contribution to the agave fructan knowledge of the Mexican agave diversity.

This review aims to assess the effects of dietary supplementation with inulin-type fructans (ITF) compared with no supplementation on cardiovascular disease risk factors in adults and assess the quality of trial reporting using the Consolidated Standards of Reporting Trials (CONSORT) and CONSORT for abstract (CONSORT-A) checklists.We will search randomised controlled trials (RCTs) in MEDLINE, EMBASE, CINAHL, Emcare, AMED and the Cochrane Database of Systematic Reviews from inception to 31 March 2022, without any language restrictions. The RCTs need to administer ITF in adults for at least 2 weeks and assess effects on at least one cardiovascular risk factor. We will exclude RCTs that (1) assessed the postprandial effects of ITF; (2) included pregnant or lactating participants; (3) enrolled participants undergoing treatment that might affect the response to ITF. We will assess the study risk of bias (RoB) using V.2 of the Cochrane RoB tool for RCTs (RoB 2) and the certainty of the evidence using the Grading of Recommendations, Assessment, Development and Evaluations (GRADE) approach. We will pool data using a random-effects model. We will use the χEthics approval is not required for secondary analysis of already published data. We will publish the reviews in a peer-review journal.CRD42019136745.

Ryegrass (Lolium perenne L.) regrowth after defoliation results from the mobilization of sugar reserves (mainly fructans) and, simultaneously, the efficient lateral transport of sucrose toward growing tissues. However, as for grasses overall, it is not yet known if the induction of this transport is solely linked to the sugar demand of growing tissues via the modification of sugar content at the tissue or cellular level or if it could be triggered by a wounding signal due to the defoliation itself. Ryegrass plants were therefore submitted to total or partial defoliation, pinning of the leaf blades to simulate wounding, or to leaf spraying with 100 μM methyl jasmonate (MeJA), a phytohormone related to wounding. As a response to total or partial defoliation, fructans were mobilized, and the expression of the sucrose lateral transporter LpSUT1 was induced. This highlights an efficient intra-plant compensatory partitioning of sugar resources between defoliated and intact tillers, resulting in the adaptation to regrow after moderate to severe defoliation. The MeJA treatment strongly decreased fructan content. Pinning and especially MeJA largely and quickly increased sucrose content and LpSUT1 transcript levels in leaf sheaths and elongating leaf bases, suggesting a direct effect of wounding on the upregulation of the sucrose lateral transporter. The overall results suggest that sucrose transport capacity and fructan degradation are induced by defoliation through the modification of source-sink relationships for sugars at the plant level and are mediated by phytohormones associated with wounding, such as jasmonates.

To provide theoretical guidance and technical support for oat production in dry farming area, we exa-mined the effects of moisture and humic acid (HA) on the accumulation and distribution of non-structural carbohydrates (NSC) in different organs of oat, as well as the mechanism of sugar metabolism and grain weight formation of oat. Two oat cultivars, 'Mengnong Dayan 1' and 'Neiyan 5', were used as experimental material. HA and clean water (CK) were foliar sprayed under dry framing (without irrigation) and limited irrigation (irrigated twice at jointing and heading stage). The dynamics of NSC components in stems, leaves and panicles, as well as the changes of carbon metabolism-related enzyme activities at different growth stages of oats after flowering were measured. Results showed that the trend of the contents of NSC in stems, leaves and panicles, in both two oat cultivars increased firstly and then decreased with the prolong of post-anthesis time. The contents of NSC in different organs were similar between two oat cultivars. Under irrigation treatment, the fructan content in panicle for Mengnong Dayan 1 of HA was higher than that of CK, with magnitude of enhancement being significantly greater than that corresponding treatment under dry farming. Under irrigation condition, the fructan, invertase activities in leaves and grain weight per panicle for Mengnong Dayan 1 of HA were increased by 27.1%, 30.6% and 55.9% compared with CK, respectively. Further, the increase trend under irrigation was stronger than that under dry farming condition. For Mengnong Dayan 1, the 1000-grain weight and grain weight per panicle were positively correlated with the content of fructan in leaves. In conclusion, the synergistic effect of moisture and humic acid could effectively regulate the accumulation of oat fructans and the activities of carbon metabolic enzymes, consequently promoting the formation of yield.探讨水分和腐植酸(HA)对燕麦不同器官非结构性碳水化合物(NSC)积累与分配的影响,进一步明确水分和HA对燕麦糖代谢和粒重形成的作用机制,可为旱作地区燕麦的推广种植提供理论指导和技术支撑。试验以‘蒙农大燕1号'和‘内燕5号'两个燕麦品种为材料,分别在旱作(无灌溉)和有限灌溉(拔节期和抽穗期每次灌水60 mm)两个水分条件下喷施HA与清水(CK),研究燕麦开花后不同时期NSC组分在茎、叶、穗中的动态变化以及叶片中碳代谢相关酶活性的变化。结果表明: 两个燕麦品种茎、叶、穗中的NSC组分含量均随开花后时间的延长先升高后降低,且两品种各器官中的NSC组分含量大致相同;与CK相比,在灌水条件下喷施HA后蒙农大燕1号穗部的果聚糖含量提升幅度明显大于旱作条件;喷施HA后蒙农大燕1号叶片中果聚糖外水解酶和转化酶活性分别显著提高了27.1%和30.6%,单穗粒重显著提高了55.9%,且与旱作条件下相比提高幅度更大;蒙农大燕1号籽粒千粒重和单穗粒重与叶片果聚糖含量呈显著正相关关系。综上,水分和腐植酸协同作用可以有效调节燕麦果聚糖的积累及主要代谢酶活性,从而提高千粒重和单穗粒重,促进产量形成。.

[Background] The consumption of wheat/gluten is associated with adverse reactions for human health. Gluten and fructans are identified as the major compounds triggering and worsening adverse reactions to wheat, which are increasing, and as a consequence, avoidance of gluten/wheat is the common strategy of many individuals of the western population. Although bread is a product of daily consumption, there is a lack of information on the gluten and fructan contents and the influence of artisanal or industrial processes., [Objective] The aim of this study is to carry out a comparative characterization between artisan bakeries and hypermarkets in Spain for gluten and fructan contents in daily sold breads., [Design] A total of 48 types of bread highly consumed in Spain sold in artisan bakeries (long fermentation) and hypermarkets (short fermentations) were selected for comparing the gluten and fructan contents. Methods such as reverse phase-high performance liquid chromatography (RP-HPLC), R5 monoclonal antibody (moAb), and fructans protocols were used for the quantification of these compounds., [Results] Great variation for the content of gluten and fructans has been found between all bread categories. Although breads produced using long fermentation (artisan bakeries) contain significantly lower gluten, they have higher fructans than those using short fermentations (hypermarkets). Durum wheat breads had the lowest content of gluten. Moreover, spelt breads from artisan bakeries had the lowest content of fructans but not those from hypermarkets., [Discussion] In this study, we report the comparative characterizarion of the breads of the Spanish market. These food products presented variation in the amount of gluten and fructans, ligated in most of the cases to the nature of the providers: artisan bakeries against hypermarkets. Depending on the type of bread, the differences for the daily consumption of gluten and fructan can be 4.5 and 20 times, respectively., [Conclusions] We found strong differences for gluten and fructan contents among breads. These information may contribute to designing strategies to improve the management of gluten and fructans in bread., This research was supported by grant PID2019-110847RB-I00 funded by MCIN/AEI/10.13039/501100011033 and grant P20_01005 funded by Consejería de Transformación Económica, Industria, Conocimiento y Universidades (Junta de Andalucía), and ‘ERDF A way of making Europe’ by the ‘European Union’

Dietary fibers such as fructans can protect the intestinal epithelial barrier integrity, but the mechanisms underlying this protection are not completely understood. We aimed to study the protective effect of β(2→1)-β(2→6) branched graminan-type fructans (GTFs) on gut epithelial barrier function that was disrupted by three different agents which impact the barrier function

Background Levan is a well-known homopolymer of fructose composed predominantly of β-(2, 6) fructofuranosyl linkages in the backbone with occasional β-(2, 1) linkages in the branch chains with varied applications. However, high production cost due to low yield of microbial levan has become a bottleneck for its practical applications. Furthermore, factors affecting the molecular mass of the synthesized levan by Leuconostoc spp. during prolonged cultivation is not fully elucidated. Methods The cultivation condition for Leuconostoc citreum BD1707 to synthesize levan was optimized by single-factor experiments and subsequently with response surface methodology (RSM). The average molecular weight (Mw) of levan synthesized by the strain L.citreum BD1707 under the optimized cultivation conditions was monitored by high-performance size exclusion chromatography (HPSEC). Finally, the enzyme with levan-degrading activity was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results The levan yield of BD1707 reached 34.86 g/L with a corresponding productivity of 7.47 g/L/d under the optimal cultivation conditions deduced by RSM, i.e., cultivation at 26 °C and 200 rpm for 112 h in tomato juice supplemented with 172 g/L sucrose with an initial pH value of 6.12. The Mw of levan reached a peak value of 2.320 × 107 Da at 6 h of cultivation under the optimized cultivation conditions and then gradually decreased to 8.809 × 106 Da after 120 h of cultivation. Conclusion The levan yield of the strain L.citreum BD1707 could be sufficiently enhanced via cultivation condition optimization. The decrease in molecular mass of the synthesized levan was attributed predominantly to the hydrolytic activity of levansucrase secreted by L.citreum BD1707 during cultivation, with an estimated Mw of 130 KD by SDS-PAGE, while the effect of acid hydrolysis could be nearly neglected.

We profiled the grain oligosaccharide content of 154 two-row spring barley genotypes and quantified 27 compounds, mainly inulin- and neoseries-type fructans, showing differential abundance. Clustering revealed two profile groups where the ‘high’ set contained greater amounts of sugar monomers, sucrose, and overall fructans, but lower fructosylraffinose. A genome-wide association study (GWAS) identified a significant association for the variability of two fructan types: neoseries-DP7 and inulin-DP9, which showed increased strength when applying a novel compound ratio-GWAS approach. Gene models within this region included three known fructan biosynthesis genes (fructan:fructan 1-fructosyltransferase, sucrose:sucrose 1-fructosyltransferase, and sucrose:fructan 6-fructosyltransferase). Two other genes in this region, 6(G)-fructosyltransferase and vacuolar invertase1, have not previously been linked to fructan biosynthesis and showed expression patterns distinct from those of the other three genes, including exclusive expression of 6(G)-fructosyltransferase in outer grain tissues at the storage phase. From exome capture data, several single nucleotide polymorphisms related to inulin- and neoseries-type fructan variability were identified in fructan:fructan 1-fructosyltransferase and 6(G)-fructosyltransferase genes. Co-expression analyses uncovered potential regulators of fructan biosynthesis including transcription factors. Our results provide the first scientific evidence for the distinct biosynthesis of neoseries-type fructans during barley grain maturation and reveal novel gene candidates likely to be involved in the differential biosynthesis of various types of fructan in barley.

Levan is an exopolysaccharide produced by Bacillus licheniformis (strain FRI MY-55) that shows promising pharmacological activity. Phosphorylation is a chemical modification that can increase the biological and antioxidant properties of levan. In this study, levan was phosphorylated by microwave-assisted synthesis to achieve a degree of substitution of 0.29. The hydroxyl radical scavenging activity of microwave-assisted phosphorylated levan (microwave P) increased significantly (6-fold) over native levan; this activity was only slightly lower than vitamin C. Other free radical scavenging and reducing power tests revealed that Microwave P activity was increased by 30–40%. Microwave P inhibited the proliferation of HCT-116 and A549 cancer cell lines more readily than native levan with an IC50 of 1.03 mg/mL and 1.38 mg/mL for HCT-116 and A549 cells, respectively. Cells treated with native levan and its derivatives remained in the sub-G1 phase according to cell cycle analysis, whereas Microwave P treatment increased the proportion of cells undergoing apoptosis. Furthermore, Microwave P effectively upregulated pro-apoptosis marker Bax and downregulated anti-apoptosis marker Bcl-2, in addition to inducing the expression of caspase-9 and caspase-3. These findings show that levan phosphorylated via microwave-assisted synthesis showed increased antioxidant and antitumor activity over native levan or levan phosphorylated via traditional long-term heating. In particular, Microwave P possesses antiproliferative activity and can induce apoptosis through mitochondrial pathways in cancerous cells.

Levan, a β-2,6-glycosidic linked fructan, is a promising alternative for the inulin dominated fructan market. Although levan is already used in some cosmetic products, the commercial availability of the fructan is still limited. Here we show that Gluconobacter japonicus LMG 1417 is a potent levan-forming organism and a promising platform for the industrial production of levan. The levansucrase LevS1417, which is produced by G. japonicus LMG 1417 and secreted by a signal-peptide-independent pathway, exhibited extraordinary high activity (4726 ± 821 U mg−1 at 50 °C). A cell-free levan production based on the supernatant of the investigated strain led to a final levan yield of 157.9 ± 7.6 g L−1. The amount of secreted levansucrase was more than doubled by plasmid-mediated homologous overproduction of LevS1417 in G. japonicus LMG 1417. Accordingly, the space-time yield of cell-free levan production was doubled using the plasmid-bearing mutant.

Streptococcus mutans, the primary etiologic agent of human dental caries, and a variety of oral Streptococcus and Actinomyces spp. synthesize high molecular mass homopolymers of fructose (fructans) with predominantly β2,1- (inulins) or β2,6-linkages (levans). The ability of S. mutans to degrade fructans contributes to the severity of dental caries. The extracellular product of

Inulin is an important reserve polysaccharide in Asteraceae plants, and is also widely used as a sweetener, a source of dietary fibre and prebiotic. Nevertheless, a lack of genomic resources for inulin-producing plants has hindered extensive studies on inulin metabolism and regulation. Here, we present chromosome-level reference genomes for four inulin-producing plants: chicory (Cichorium intybus), endive (Cichorium endivia), great burdock (Arctium lappa) and yacon (Smallanthus sonchifolius), with assembled genome sizes of 1.28, 0.89, 1.73 and 2.72 Gb, respectively. We found that the chicory, endive and great burdock genomes were shaped by whole genome triplication (WGT-1), and the yacon genome was shaped by WGT-1 and two subsequent whole genome duplications (WGD-2 and WGD-3). A yacon unique whole genome duplication (WGD-3) occurred 5.6-5.8 million years ago. Our results also showed the genome size difference between chicory and endive is largely due to LTR retrotransposons, and rejected a previous hypothesis that chicory is an ancestor of endive. Furthermore, we identified fructan-active-enzyme and transcription-factor genes, and found there is one copy in chicory, endive and great burdock but two copies in yacon for most of these genes, except for the 1-FEH II gene which is significantly expanded in chicory. Interestingly, inulin synthesis genes 1-SST and 1-FFT are located close to each other, as are the degradation genes 1-FEH I and 1-FEH II. Finally, we predicted protein structures for 1-FFT genes to explore the mechanism determining inulin chain length.

We assessed the extent to which fructans from various sources and added in various forms (raw materials in diet alone or incorporated into a strawberry matrix) differ in their effectiveness towards selected parameters related to bone health under calcium hypoalimentation in growing female Wistar rats. The aim of this study was to evaluate the levels of selected parameters involved in calcium metabolism, in response to a 12-week restriction of Ca intake: serum ions (Ca, Mg, P); the activity of alkaline phosphatase—using a BS 120 analyzer; the markers of bone turnover (osteocalcin, CTX; using a Rat-MidTMOsteocalcinEIA Kit and RatLapsTMEIA, respectively); and the bone mineral content (BMC) and density (BMD), using a Norland Excell Plus Densitometer. Among the examined markers, the CTX concentration increased dramatically under calcium hypoalimentation. The presence of Jerusalem artichoke (independently of the form of addition) and yacon root powder (with strawberry sorbet matrix) in the rats’ diet led to a significantly lower CTX concentration than was observed in the low-calcium control group. The type of fructan influenced the bone mass content. When fructan was added to the low-calcium diet as an ingredient of sorbet, it exerted more pronounced effects on the biochemical parameters of bone metabolism than when added alone, in the growing-female-rat model.

The study focused on the evaluation of the possibility of using a levan-rich digestate extract in the production of safe and functional body wash cosmetics. Model shower gels were designed and formulated on the basis of raw materials of natural origin. Prepared prototypes contained various extract concentrations (16.7; 33; 50%). A gel without extract was used as a reference. The samples were evaluated for their safety in use and functionality. The results showed that the use of high-concentration levan-rich digestate extract in a shower gel resulted in a significant reduction in the negative impact on the skin. For example, the zein value decreased by over 50% in relation to the preparation without the extract. An over 40% reduction in the emulsifying capacity of hydrophobic substances was also demonstrated, which reduces skin dryness after the washing process. However, the presence of the extract did not significantly affect the parameters related to functionality. Overall, it was indicated that levan-rich digestate extract can be successfully used as a valuable ingredient in natural cleansing cosmetics.

Microbial levansucrases (LSs, EC 2.4.1.10) have been widely studied for the synthesis of β-(2,6)-fructans (levan) from sucrose. LSs synthesize levan-type fructo-oligosaccharides, high-molecular-mass levan polymer or combinations of both. Here, we report crystal structures of LS from the G

Conventional fossil fuel-based packaging materials often brings of food safety and serious environmental pollution. It is significant to develop an environmentally-friendly packaging material. In this work, a levan-chitosan (LE/CS) blend film was fabricated via the solution casting method. The films were evaluated by Fourier transform infrared spectroscopy and X-ray diffraction, indicating the formation of hydrogen bonds between chitosan and levan. The mechanical properties of LE/CS films demonstrated a mechanical strength higher than CS films, and the best tensile strength appeared at a ratio of LE/CS (1:1) up to 18.78 ± 0.73 MPa. The addition of levan caused a significant increase in absorption of UV light with a reduction in swelling water of the blend films from 29.13 ± 0.53 % of chitosan film to 2.07 ± 0.27 % of LE/CS (1:1) film. A higher contact angle and lower WVP were observed for LE/CS blend films. LE/CS blend films were then used as packaging material for fresh pork and were well maintained the qualities. The study suggested that the new blend film might have a good prospect as a food packaging material.

High fructan content in the grain of cereals is an important trait in agriculture such as environmental resilience and dietary fiber food production. To understand the mechanism in determining final grain fructan content and achieve high fructan cereal, a cross breeding strategy based on fructan synthesis and hydrolysis activities was set up and have achieved barley lines with 11.8% storage fructan in the harvested grain. Our study discovered that high activity of fructan hydrolysis at later grain developmental stage leads to the low fructan content in mature seeds, simultaneously increasing fructan synthesis at early stage and decreasing fructan hydrolysis at later stage through crossing breeding is an efficient way to elevate grain diet-fiber content. A good correlation between fructan and beta glucans was also discovered with obvious interest. Field trials showed that the achieved high fructan barley produced over seven folds higher fructan content than control barley and pull carbon-flux to fructan through decreasing fructan hydrolysis without disruption starch synthesis will probably not bring yield deficiency.

The aim of this work has been to establish and validate a simple and efficient method to detect the concentration of inulin-type fructan CPA from the roots of Codonopsis pilosula (Franch.) Nannf. in biosamples, and then apply it to evaluate the pharmacokinetics behavior, distribution character in tissue and excretion in mice. In this work, fluorescein isothiocyanate (FITC) was used to label CPA. Then FCPA was intravenously and orally administered to mice at different doses. In both i.v and p.o administration, FCPA concentration slowly declined in the circulatory system with a much longer

Cultivation of crops in salt-affected soils is a major challenge for growers. Despite the use of multiple amendments, salinity stresses adversely affect the crops to some extent. On the other hand, imbalance in the use of boron (B) as a nutrient also creates toxicity. Mismanagement of B fertilizer application decreases the growth and yield of crops. It is necessary to study in depth the adverse effects of salinity and B toxicity. This is why the current research work was conducted in a glass house at Murdoch University, Perth, Australia. The aim of study was to investigate the influence of salinity and B toxicity on carbohydrate partitioning, growth, and ionic composition of two Australian wheat varieties. There were four treatments, i.e., control, high B (15 kg ha−1), salinity (15 dS m−1), and B + salinity. The results showed that the salt-tolerant Halberd (HB) variety accumulated more Na+, B, and Cl− in their leaf sheath and kept the leaf blades free of these toxic ions as compared to the sensitive variety Westonia (WS). Water-soluble carbohydrate (WSC; i.e., glucose, sucrose, fructose, and fructans) concentration increased in response to individual as well as combined constrains of soil salinity and toxic B in the leaf blade of both tolerant and sensitive wheat varieties, but the increase was higher in the tolerant variety as compared to the sensitive one. The concentration of WSCs in leaf sheath of the salt-tolerant wheat variety was increased in response to stress conditions, but those remained low in salt-sensitive ones. Therefore, the salt-tolerant HB genotype was found to be a good source for future wheat breeding programs or to be grown by farmers in B toxic, saline, and B toxic–saline conditions.

Resumen Los fructanos son carbohidratos de estructura química diversa distribuidos en diferentes taxa. En las plantas, además de constituir una importante fuente de carbono, han sido asociados con la tolerancia a diferentes tipos de estrés. La biosíntesis de estos compuestos se lleva a cabo por la acción de las enzimas fructosiltransferasas, mientras que su degradación es mediada por las exohidrolasas fructosílicas, ambos tipos forman parte de la familia 32 de las enzimas hidrolasas glicosídicas. Las exohidrolasas fructosílicas son exoenzimas que liberan secuencialmente residuos de fructosa terminales de los fructanos para hacerlos disponibles como fuente de carbono en diferentes procesos celulares o bien moléculas de señalización. En esta revisión se hace una descripción de los fructanos y las exohidrolasas fructosílicas en algunas plantas importantes para el consumo humano y para el aprovechamiento industrial, con un enfoque particular en el género Agave, específicamente en A. tequilana. Concluimos, que el estudio de las exohidrolasas fructosílicas en agaves podría ser útil en varias aplicaciones biotecnológicas como en la hidrólisis de estructuras de fructanos no digeridos durante la etapa de cocción del agave en el proceso de producción de tequila. Sin embargo, el primer paso es conocer la funcionalidad de estas enzimas, lo que podría facilitar su incorporación en diferentes procesos biotecnológicos. Abstract Fructans are carbohydrates of diverse chemical structure that are distributed in different taxa. In plants, in addition to be an important source of carbon, they have been associated with tolerance to different types of stress. The biosynthesis of these compounds is carried out by the action of enzymes called fructosyltransferases, while their degradation is mediated by fructan exohydrolases, both types are part of the 32 family of glycosidic hydrolase enzymes. Fructan exohydrolases are exo-enzymes that sequentially release terminal fructose residues from fructans to make them available as carbon sources for different cellular processes or as signaling molecules. In this review, a description is made of fructans, y of fructosyl exohydrolases in some plants important for human consumption or for industrial use, such as the Agave genus, specifically in A. tequilana. We conclude that the study of fructan exohydrolases in agaves could be useful in various biotechnological applications, for example, in the hydrolysis of undigested fructan structures during the agave cooking stage in the tequila production process. However, the first step is to determinate the enzymatic activity in which they are involved, for its posterior inclusion in biotechnology processes. Resumo Os frutanos são carboidratos de estrutura química diversificada distribuídos em diferentes táxons. Nas plantas, além de serem uma importante fonte de carbono, eles têm sido associados à tolerância a diferentes tipos de estresse. A biossíntese desses compostos é realizada pela ação das enzimas fructosiltransferases, enquanto sua degradação é mediada por frutossil exohidrolases, ambos os tipos fazem parte da família 32 de enzimas glicosídicas da hidrolase. As exo-hidrolases de frutossil são exo-enzimas que liberam seqüencialmente resíduos terminais de frutose dos frutanos para torná-los disponíveis como fonte de carbono em diferentes processos celulares, ou como sinalizadores. Nesta revisão, é feita uma descrição dos frutanos e das frutossil exohidrolases em algumas plantas importantes para o consumo humano ou para uso industrial, como o gênero Agave, especificamente no A. tequilana. Concluímos que o estudo de frutossil exo-hidrolases em agaves pode ser útil em várias aplicações biotecnológicas, por exemplo, na hidrólise de estruturas de frutano não digeridas durante a fase de cozimento da agave no processo de produção de tequila. Para isso, o primeiro passo é conhecer o tipo de atividade enzimática que eles desempenham, para sua posterior inclusão no campo da biotecnologia.

Asparagus (Asparagus officinalis L.) accumulates inulin and inulin neoseries-type fructans in root, which are synthesized by three fructosyltransferases-sucrose:sucrose 1-fructosyltransferase (1-SST, EC 2.4.1.99), fructan:fructan 1-fructosyltransferase (1-FFT, EC 2.4.1.100), and fructan:fructan 6G-fructosyltransferase (6G-FFT, EC 2.4.1.243). Fructans in roots are considered as energy sources for emerging of spears, and it has been demonstrated that a gradual decrease in root fructan content occurs during the spear harvesting season (budding and shooting up period). However, the roles of certain three fructosyltransferases during the harvest season have not yet been elucidated. Here, we investigated the variation in enzymatic activities and gene expression levels of three fructosyltransferases and examined sugar contents in roots before and during the spear harvest period. Two cDNAs, aoft2 and aoft3, were isolated from the cDNA library of roots. The respective recombinant proteins (rAoFT2 and rAoFT3), produced by Pichia pastoris, were characterized: rAoFT2 showed 1-FFT activity (producing nystose from 1-kestose), whereas rAoFT3 showed 1-SST activity (producing 1-kestose from sucrose). These reaction profiles of recombinant proteins were similar to those of native enzymes purified previously. These results indicate that aoft2 and aoft3 encoding 1-FFT and 1-SST are involved in fructan synthesis in roots. A gradual downregulation of fructosyltransferase genes and activity of respective enzymes was observed in roots during the harvest period, which also coincided with the decrease in fructooligosaccharides and increase in fructose due to fructan exohydrolase activity. These findings suggest that downregulation of fructosyltransferases genes during harvest time may contribute to efficient degradation of fructan required for the emergence of spears.