Search Results (28)

Fructooligosaccharides (FOS) are short fructans with different degrees of polymerization (DP) and bonds in their structure, generated by the distinct activities of fructosyltransferase enzymes, which produce distinct types of links. FOS are in high demand on the market, mainly because of their prebiotic effects. In recent years, depending on the link type in the FOS structure, prebiotic activity has been shown to be increased. Studies on β-fructanofuranosidases (Ffasa), enzymes with fructosyltransferase activity in yeasts, have reported the production of ¹F-FOS, ⁶F-FOS, and ⁶G-FOS. The aims of this investigation were to evaluate the capability of fifteen different yeasts to grow in Agave sp. juices and to determine the potential of these juices as substrates for FOS production. Additionally, the research aimed to corroborate and analyze the fructosyltransferase activity of enzymatic extracts obtained from agave yeasts by distinct induction media and to identify the role and optimal parameters (time and sucrose and glucose concentrations) for FOS and disaccharides production through Box–Behnken designs. To carry out such a task, different techniques were employed: FT-IR, TLC, and HPAEC-PAD. We found two yeasts with fructosyltransferase activity, P. kudriavzevii ITMLB97 and C. lusitaniae ITMLB85. In addition, within the most relevant results, the production of the FOS 1-kestose, 6-kestose, and neokestose, as well as disaccharides inulobiose, levanobiose, and blastose, molecules with potential applications, was determined. Overall, FOS production requires suitable yeast species, which grow in a medium under optimal conditions, from which microbial enzymes with industrial potential can be obtained. Full article

Distinguishing between endo- and exo-type enzymes within the glycoside hydrolase (GH) classification presents significant challenges. Traditional methods, often based on endpoint activity measurements, do not capture the full range of products generated, leading to inconsistencies in classification. Not all exo-acting fructanases and glucanases produce monosaccharides (like fructose or glucose), while endo-acting enzymes do not solely produce higher-degree polymerization oligosaccharides. In practice, both enzyme types can yield a variety of products throughout the reaction, complicating classification efforts. To address these challenges, we propose a refined nomenclature system for GH enzymes, including fructanases and glucanases, based on good practices and initial product formation. This system classifies enzymes into four categories for each type: Fr, Fn, Fn,n+1 and F1 for fructanases, and Gr, Gn, Gn,n+1 and G1 for glucanases, based on their mode of action (endo- or exo-based) and initial product profiles. Our refined nomenclature system will advance enzyme structure–function research and support the production and application of fructan and glucan oligosaccharides as prebiotics, priming agents, and potential signaling molecules in microbe–microbe and plant–microbe interactions. Ultimately, this system could benefit agronomy and the food industry, contributing to health improvements. Full article

Inulin is a carbohydrate that belongs to fructans; due to its health benefits, it is widely used in the food and pharmaceutical industries. In this research, cabuya (Agave americana) was employed to obtain inulin by pulsed electric field-assisted extraction (PEFAE) and FTIR analysis confirmed its presence. The influence of PEFAE operating parameters, namely, electric field strength (1, 3 and 5 kV/cm), pulse duration (0.1, 0.2 and 0.5 ms), number of pulses (10,000, 20,000 and 40,000) and work cycle (20, 50 and 80%) on the permeabilization index and energy expenditure were tested. Also, once the operating conditions for PEFAE were set, the temperature for conventional extraction (CE) and PEFAE were defined by comparing extraction kinetics. The cabuya meristem slices were exposed to PEFAE to obtain extracts that were quantified, purified and concentrated. The inulin was isolated by fractional precipitation with ethanol to be characterized. The highest permeabilization index and the lowest energy consumption were reached at 5 kV/cm, 0.5 ms, 10,000 pulses and 20%. The same extraction yield and approximately the same amount of inulin were obtained by PEFAE at 60 °C compared to CE at 80 °C. Despite, the lower amount of inulin obtained by PEFAE in comparison to CE, its quality was better because it is mainly constituted of inulin of high average polymerization degree with more than 38 fructose units. In addition, TGA analyses showed that inulin obtained by PEFAE has a lower thermal degradation rate than the obtained by CE and to the standard. Full article

Agave tequilana stems store fructan polymers, the main carbon source for tequila production. This crop takes six or more years for industrial maturity. In conducive conditions, agave wilt disease increases the incidence of dead plants after the fourth year. Plant susceptibility induced for limited photosynthates for defense is recognized in many crops and is known as “sink-induced loss of resistance”. To establish whether A. tequilana is more prone to agave wilt as it ages, because the reduction of water-soluble carbohydrates in roots, as a consequence of greater assembly of highly polymerized fructans, were quantified roots sucrose, fructose, and glucose, as well as fructans in stems of agave plants of different ages. The damage induced by inoculation with Fusarium solani or F. oxysporum in the roots or xylem bundles, respectively, was recorded. As the agave plant accumulated fructans in the stem as the main sink, the amount of these hexoses diminished in the roots of older plants, and root rot severity increased when plants were inoculated with F. solani, as evidence of more susceptibility. This knowledge could help to structure disease management that reduces the dispersion of agave wilt, dead plants, and economic losses at the end of agave’s long crop cycle. Full article

Fructans are fructose-based polymers, defined as fructooligosaccharides (FOS), when they possess a short chain. These molecules are highly appreciated in the food and pharmaceutical international market and have an increasing demand worldwide, mainly for their prebiotic activity and, therefore, for all their health benefits to those who consume them constantly. Thus, new natural or alternative FOS production systems of industrial scale are needed. In this regard, microorganisms (prokaryotes and eukaryotes) have the potential to produce them through a wide and diverse number of enzymes with fructosyltransferase activity, which add a fructosyl group to sucrose or FOS molecules to elongate their chain. Microbial fructosyltransferases are preferred in the industry because of their high FOS production yields. Some of these enzymes include levansucrases, inulosucrases, and β-fructofuranosidases obtained and used through biotechnological tools to enhance their fructosyltransferase activity. In addition, characterizing new microorganisms with fructosyltransferase activity and modifying them could help to increase the production of FOS with a specific degree of polymerization and reduce the FOS production time, thus easing FOS obtention. Therefore, the aim of this review is to compile, discuss, and propose new perspectives about the microbial potential for FOS production through enzymes with fructosyltransferase activity and describe the modulation of FOS production yields by exogenous stimuli and endogenous modifications. Full article

Fructans serve as the primary form of storage carbohydrate in cool-season grasses, but little is known about potential differences in ruminal fermentation of fructans between cattle and sheep. An ex vivo study was conducted to evaluate species differences in fructan catabolism. Buffered media containing ground orchardgrass (Dactylis glomerata L.) substrate was inoculated with uncultivated rumen microbiota obtained from cattle and sheep (n = 4 species⁻¹). Fructan profiles were monitored over the incubation period (8 h; 39 °C) using high-performance anion-exchange chromatography coupled to pulsed amperometric detection (HPAEC-PAD). In both species, disappearance of long-chain fructans (degree of polymerization [DP] > 8) was evident by 2 h of incubation (p < 0.01), whereas short-chain fructans (DP 4–8) increased from 0 to 2 h prior to subsequent degradation (p < 0.01). However, the overall rate of long-chain fructan catabolism was greater in bovine versus ovine fermentations, particularly between 2 and 4 h (p < 0.01). Additionally, rapid utilization of short-chain fructans occurred from 2 to 4 h in bovine fermentations, but was delayed in ovine fermentations, with substantial degradation occurring only after 4 h of incubation (p < 0.01). These results indicate that rumen microbiota of cattle may have a greater capacity for fructan degradation. Full article

The effect of chemical modification of fractions of native agave fructans (NAF), high performance (HPAF), and a high degree of polymerization (HDPAF) by lauroylation, acetylation, and succinylation reactions on their prebiotic activity, antibacterial properties were evaluated and survival of L. paracasei in a simulated gastrointestinal system. The characterization of the reactions was confirmed by NMR and FTIR. The lauroylated and succinylated fructan fractions showed higher antibacterial activity against pathogenic bacteria such as Escherichia coli, Enterococcus faecalis and Staphylococcus aureus than the unmodified ones. Analyses with L. paracasei showed that the acetylated fructan fractions had a greater prebiotic effect, and simulated gastrointestinal tests demonstrated that the acetylated and succinylated fractions favored the survival of L. paracasei during the gastrointestinal phase. The effect of modifying the agave fructans fractions on the evaluated properties depended on the structure, size, and polarity of each incorporated functional group, as well as the degree of polymerization and substitution of each fraction. These results show that the chemical modification of the fructan fractions analyzed improves their functional properties, offering an alternative in the food and pharmaceutical industry. Full article

Prebiotic effects have been attributed to agave fructans through bacterial and yeast fermentations, but there are few reports on their use as raw materials of a carbon source. Kefir milk is a fermented drink with lactic acid bacteria and yeast that coexist in a symbiotic association. During fermentation, these microorganisms mainly consume lactose and produce a polymeric matrix called kefiran, which is an exopolysaccharide composed mainly of water-soluble glucogalactan, suitable for the development of bio-degradable films. Using the biomass of microorganisms and proteins together can be a sustainable and innovative source of biopolymers. In this investigation, the effects of lactose-free milk as a culture medium and the addition of other carbon sources (dextrose, fructose, galactose, lactose, inulin and fructans) in concentrations of 2, 4 and 6% w/w, coupled with initial parameters such as temperature (20, 25 and 30 °C), % of starter inoculum (2, 5 and 10% w/w) was evaluated. The method of response surface analysis was performed to determine the optimum biomass production conditions at the start of the experiment. The response surface method showed that a 2% inoculum and a temperature of 25 °C were the best parameters for fermentation. The addition of 6% w/w agave fructans in the culture medium favored the growth of biomass (75.94%) with respect to the lactose-free culture medium. An increase in fat (3.76%), ash (5.57%) and protein (7.12%) content was observed when adding agave fructans. There was an important change in the diversity of microorganisms with an absence of lactose. These compounds have the potential to be used as a carbon source in a medium culture to increase kefir granule biomass. There was an important change in the diversity of microorganisms with an absence of lactose, where the applied image digital analysis led to the identification of the morphological changes in the kefir granules through modification of the profile of such microorganisms. Full article

Commercial fructans (inulin and oligofructose) are generally obtained from crops such as chicory, Jerusalem artichoke or agave. However, there are agricultural by-products, namely asparagus roots, which could be considered potential sources of fructans. In this work, the fructans extracted from asparagus roots and three commercial ones from chicory and agave were studied in order to compare their composition, physicochemical characteristics, and potential health effects. Asparagus fructans had similar chemical composition to the others, especially in moisture, simple sugars and total fructan contents. However, its contents of ash, protein and phenolic compounds were higher. FTIR analysis confirmed these differences in composition. Orafti^®GR showed the highest degree of polymerization (DP) of up to 40, with asparagus fructans (up to 25) falling between Orafti^®GR and the others (DP 10–11). Although asparagus fructan powder had a lower fructan content and lower DP than Orafti^®GR, its viscosity was higher, probably due to the presence of proteins. The existence of phenolic compounds lent antioxidant activity to asparagus fructans. The prebiotic activity in vitro of the four samples was similar and, in preliminary assays, asparagus fructan extract presented health effects related to infertility and diabetes diseases. All these characteristics confer a great potential for asparagus fructans to be included in the prebiotics market. Full article

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. Full article

The gut microbiota is implicated in diverse interactions affecting human health. The present study reports a randomized, double-blind, placebo-controlled clinical study conducted by administering a new synbiotic formulation composed of two Lactobacillus strains (L. plantarum and L. acidophilus) and one Bifidobacterium strain (B. animalis subsp. lactis) and two types of fructans (fructo-oligosaccharides with a degree of polymerization of 3–5 and inulin-type fructans with 10 DP). The effects of this synbiotic were evaluated on healthy subjects for 28 days and the maintenance of its efficacy was evaluated at the end of a follow-up period of 28 days. The synbiotic treatment contributes to higher biodiversity of the gut microbiota, increasing the community richness with respect to the group with the prebiotics alone and the placebo group. Its positive effect is also reflected in the variation of microbial community structure favoring the beneficial short-chain fatty acids bacterial producers. The amelioration of the health status of the subjects was also established by the reduction of common infectious disease symptom incidence, the stimulation of the gut immune system showing a noteworthy variation of fecal β-defensin2 and calprotectin levels, and the modulation of the response of the respiratory tract’s immune system by salivary IgA as well as total antioxidant capacity biomarkers. Full article

Levansucrase LevS from Leuconostoc mesenteroides B-512F is a multidomain fructansucrase (MD-FN) that contains additional domains (ADs) to the catalytic domain. However, the understanding of the effect that these ADs have on enzyme activity remains vague. To this aim, structure-function relationship studies of these LevS ADs were performed by evaluating both biochemical properties and the enzymatic capacity of truncated versions of LevS. Joint participation of the N- and C-terminal domains is essential for stability, activity, specificity, and polymerization processes. Specifically, the N-terminal region is involved in stability, while the transition region plays an essential role in the transfructosylation reaction and polymer elongation. Based on our results, we suggest that ADs interact with each other, adopting a U-shaped topology. The importance of these ADs observed in the MD-FN of the Leuconostocaceae family is not shared by the Lactobacillaceae family. Phylogenetic analysis of LevS AD suggests that MD-FN from Lactobacillaceae and Leuconostocaceae have different evolutionary origins. This is the first study on the structure-function relationship of multidomain levansucrases from the Leuconostocaceae family. Our results point towards the functional role of AD in MD-FN and its involvement in fructan synthesis. Full article

Asparagus roots are by-products from asparagus cultivation and they could be considered one of the best sources of fructans. These polymers are interesting food ingredients for their prebiotic and immuno-stimulating characteristics. The aim of this work is to characterize the fructan profile from the roots of several asparagus varieties grown at different locations and pickled at three vegetative statuses in order to valorize these by-products as fructan source. Fructans were extracted with hot water and fractionated into three pools according to their molecular weight (MW). Their average MW was studied by HPSEC and their degree of polymerization by HPAEC. The fructan content was up to 12.5% on fresh weight basis, depending on variety and sampling date. The relative abundance of the three pools also depended on the picking moment as after the spear harvest period their total content and MW increased. The average MW of the three fractions was similar among varieties with 4.8, 8.4 and 9 sugar units, although fructans up to 30 units were identified by HPAEC. These characteristics make them similar to the commercialized Orafti^®-GR inulin, a common additive to food products. Therefore, the concept of asparagus roots as cultivation waste must be changed to a new feedstock for sustainable agriculture and industry. Full article

Plant-based systems continue to play a pivotal role in healthcare, and their use has been extensively documented. Asphodelus L. is a genus comprising various herbaceous species, known by the trivial name Asphodelus. These plants have been known since antiquity for both food and therapeutic uses, especially for treating several diseases associated with inflammatory and infectious skin disorders. Phytochemical studies revealed the presence of different constituents, mainly anthraquinones, triterpenoids, phenolic acids, and flavonoids. Although extensive literature has been published on these constituents, a paucity of information has been reported regarding the carbohydrate composition, such as fructans and fructan-like derivatives. The extraction of water-soluble neutral polysaccharides is commonly performed using water extraction, at times assisted by microwaves and ultrasounds. Herein, we reported the investigation of the alkaline extraction of root-tubers of Asphodelus ramosus L., analyzing the water-soluble polysaccharides obtained by precipitation from the alkaline extract and its subsequent purification by chromatography. A polysaccharide was isolated by alkaline extraction; the HPTLC study to determine its composition showed fructose as the main monosaccharide. FT-IR analysis showed the presence of an inulin-type structure, and NMR analyses allowed us to conclude that A. ramosus roots contain polysaccharide with an inulin-type fructooligosaccharide with a degree of polymerization of 7–8. Full article

Synbiotics aim to improve gastrointestinal health by combining pre- and probiotics. This study evaluated combinations of Bifidobacterium animalis subsp. lactis BB-12 with seven fructans: oligofructoses (OF1-OF2; low degree of polymerization (DP)), inulins (IN1-IN2-IN3; high DP) and OF/IN mixtures (OF/IN1-OF/IN2). During monoculture incubations, all fructans were fermented by BB-12 as followed from increased BB-12 numbers and increased acetate and lactate concentrations, with most pronounced fermentation for low DP fructans (OF1-OF2). Further, short-term colonic incubations for three human donors revealed that also in presence of a complex microbiota, all fructans (particularly OF1) consistently selectively enhanced the growth of BB-12. While each fructan as such already increased Bifidobacteriaceae numbers with 0.94–1.26 log(cells/mL), BB-12 co-supplementation additionally increased Bifidobacteriaceae with 0.17–0.46 log(cells/mL). Further, when co-supplemented with fructans, BB-12 decreased Enterobacteriaceae numbers (significant except for IN1-IN3). At metabolic level, all fructans decreased pH due to increased acetate and lactate production, while OF/IN2-IN1-IN2-IN3 also stimulated propionate and butyrate production. BB-12 co-supplementation further increased propionate and butyrate for OF/IN2-IN3 and IN1-IN2, respectively. Overall, combinations of BB-12 with fructans are promising synbiotic concepts, likely due to intracellular consumption of low DP-fructans by BB-12 (either present in starting product or released upon fermentation by indigenous microbes), thereby enhancing effects of the co-administered fructan. Full article