Selective enzymatic α-1,6- monoglucosylation of mogroside IIIE for the bio-creation of α-siamenoside I, a potential high-intensity sweetener.

A new compound, α-siamenoside I (α-SI), with a glucose unit selectively bound to the 6-hydroxyl group of the 24-O-ß-glucosyl moiety of mogroside IIIE by α-1,6-glucosidic bond, was bio-created by two screened cyclodextrin glycosyltransferases with a maximum yield of 59.3%. Compared to mogroside IIIE, α-SI showed a significantly increased sweetness intensity (508 times sweeter than 5% sucrose), which is superior to siamenoside I (SI), the sweetest triterpenoid saponin isolated from Siraitia grosvenorii to date. Sensory evaluation showed that the taste quality of α-SI also was obviously better than mogroside IIIE. In addition to α-SI possessing a good stability similar to that of SI, it also did not cause a significant decrease in cell viability at a concentration of 200 µg/mL and had a negative influence on islets function at 1 µM. All of these preliminarily results pave the way for promoting α-SI as a potential low-calorie sweetener.

Non-nutritive sweeteners are in concomitant with the formation of endogenous and exogenous advanced glycation end-products.

This study investigated the role of non-nutritive sweeteners in the formation of advanced glycation end-products (AGEs) and their reactive intermediates using endogenous and exogenous models. In the endogenous model, xylitol and sorbitol formed similar levels of reactive intermediates compared to sucralose. Protein-bound fluorescent AGEs, Nε-carboxymethyllysine (CML), and Nε-carboxyethyllysine (CEL) levels in the xylitol and sorbitol treatment were significantly higher compared to the sucralose treatment. In the exogenous model, sucralose treatment showed significantly higher glyoxal and fructosamine levels compared to xylitol and sorbitol, respectively. However, protein-bound fluorescent AGEs, CML, and CEL were lower in the sucralose treatment compared to other sugar treatments. The data suggest that the structure of sugar alcohols which are similar to reducing sugars may contribute to the formation of AGEs and their reactive intermediates in the endogenous model. The long-term effects of non-nutritive sweeteners consumption on AGEs formation and health implications should be verified with population studies.

Evaluation of Sweetener Synergy in Humans by Isobole Analyses.

The chemical senses and pharmaceuticals fundamentally depend on similar biological processes, but novel molecule discovery has classically been approached from vastly different vantage points. From the perspective of ingredient and flavor companies, there are countless ingredients that act via largely unknown mechanisms, whereas the pharmaceutical industry has numerous mechanisms in search of novel compounds. Mixtures of agonists can result in synergistic (superadditive) responses, which can be quantified via isobole analysis, a well-proven clinical approach in pharmacology. For the food and beverage industries, bulk (caloric) sweeteners like sugars are a key ingredient in sweetened foods and beverages, but consumers also desire products with fewer calories, which has led to the development of sweet enhancers and sweetener blends intended to achieve synergy or superadditivity. Synergistic mixtures are highly attractive targets commercially as they enable lower usage levels and enhanced efficacy. Although the psychophysical literature contains numerous prior reports of sweetener synergy, others have also noted that classical additive models fail to account for nonlinear dose-response functions. To address this shortcoming, here we systematically apply the isobole method from pharmacology to quantify the presence or absence of psychophysical synergy for binary pairs of sweeteners in a series of 15 separate experiments, each with ~100 adult volunteers (total n = 1576). Generally, these data support the hypothesis that structurally similar sweeteners acting as agonists will not synergize, whereas structurally dissimilar sweeteners binding to overlapping or distal sites can act as allosteric agonists or agonist-antagonists, respectively.

Zùsto: A new sweetening agent with low glycemic index.

BACKGROUND & AIMS: Sweetening agents are sugar substitutes with a low glycemic index, used to obtain a better glycemic control in diabetes patients. However, they also may have a role in other subjects, as a high glycemic index is thought to cause many pathological conditions. Unfortunately, not all artificial sweeteners are perceived as sweet as sugar by patients. Consumers refer often to an after taste present in foods sweetened with intensive sweeteners. The objective of this study was to explore whether Zùsto® had a low glycemic index, to replace glucose as a sweetener. METHODS: In this study, the glycemic index (GI) of a new sweetening agent, Zùsto®, is compared to that of glucose 25 g, a standard sugar-loaded drink used in the oral glucose tolerance test to detect diabetes, as primary endpoint. Zùsto® is composed of non-digestible, water soluble fibers and sweeteners. 10 healthy, female non-obese volunteers received glucose and Zùsto®, albeit by an interval of a week. Evolution of glycemia, C-peptide and insulin release was measured at different time-points after intake. RESULTS: The results show that, when calculating the mean incremental Area Under the Curve (AUC), the AUC of glucose was around five times as high as that of Zùsto®; a GI of 22 for Zùsto® was calculated. Furthermore, Zùsto® had no significant effect on the glycemia, contrary to glucose, for at least 60'. This was also the case concerning C-peptide and insulin release, but the difference lasted even for 180'. Moreover, Zùsto® was perceived as sweet by all volunteers, with no particular aftertaste. CONCLUSION: Zùsto® could be a viable alternative for fast sugars and other sweetening agents, both for diabetic patients and other subjects, requiring however a larger trial to confirm these results. CLINICALTRIALS.GOV: NCT02607345.

Non-caloric sweetener provides magnetic resonance imaging contrast for cancer detection.

BACKGROUND: Image contrast enhanced by exogenous contrast agents plays a crucial role in the early detection, characterization, and determination of the precise location of cancers. Here, we investigate the feasibility of using a non-nutritive sweetener, sucralose (commercial name, Splenda), as magnetic resonance imaging (MRI) contrast agent for cancer studies. METHODS: High-resolution nuclear-magnetic-resonance spectroscopy and MR studies on sucralose solution phantom were performed to detect the chemical exchange saturation transfer (CEST) property of sucralose hydroxyl protons with bulk water (sucCEST). For the animal experiments, female Fisher rats (F344/NCR) were used to generate 9L-gliosarcoma model. MRI with CEST experiments were performed on anesthetized rats at 9.4 T MR scanner. Following the baseline CEST scans, sucralose solution was intravenously administered in control and tumor bearing rats. CEST acquisitions were continued during and following the administration of sucralose. Following the sucCEST, Gadolinium-diethylenetriamine pentaacetic acid was injected to perform Gd-enhanced imaging for visualizing the tumor. RESULTS: The sucCEST contrast in vitro was found to correlate positively with the sucralose concentration and negatively with the pH, indicating the potential of this technique in cancer imaging. In a control animal, the CEST contrast from the brain was found to be unaffected following the administration of sucralose, demonstrating its blood-brain barrier impermeability. In a 9L glioma model, enhanced localized sucCEST contrast in the tumor region was detected while the unaffected brain region showed unaltered CEST effect implying the specificity of sucralose toward the tumorous tissue. The CEST asymmetry plots acquired from the tumor region before and after the sucralose infusion showed elevation of asymmetry at 1 ppm, pointing towards the role of sucralose in increased contrast. CONCLUSIONS: We show the feasibility of using sucralose and sucCEST in study of preclinical models of cancer. This study paves the way for the potential development of sucralose and other sucrose derivatives as contrast agents for clinical MRI applications.

Stevia-based sweeteners as a promising alternative to table sugar: The effect on Listeria monocytogenes and Salmonella Typhimurium growth dynamics.

Sugar is commonly substituted with stevia-based products in food industry and in our daily-life. This substitution results in a change in food product characteristic formula and properties that may affect the growth dynamics of food pathogenic and spoilage bacteria. This work studies the effect of table sugar (TS), laboratory sucrose (LS), commercial stevia (St) and steviol glycosides (SG) on the growth dynamics of Salmonella Typhimurium and Listeria monocytogenes. Experiments were carried out in general and minimal culture media at 3 equivalent concentration levels in terms of sweetness intensity (TS and LS at 3, 9 and 15% (w/v); St at 0.3, 0.9 and 1.5% (w/v); and SG at 0.01, 0.03 and 0.05% (w/v)). Incubation temperatures were: 4, 8 and 20°C for general media, and for minimal media 20°C. To decipher the role of these sweeteners, their concentration evolution in minimal media was determined via HPLC analysis. The results revealed slow maximum specific growth rates (µmax) of S. Typhimurium in general media with increasing concentrations of TS and LS at 20°C; and reduced maximum cell population (Nmax) at 8°C. The growth of L. monocytogenes in general culture media remains invariable independently of the sweetener added, except at 4°C. At this critical temperature, the presence of TS, LS and St seems to facilitate the growth of L. monocytogenes, presenting higher µmax values in comparison to SG and the control. Varying bacterial response to changes in media formulation suggests that further research is required, focusing on revealing the microbial dynamics in structured media, as well as in real food products.

Low calorie sweeteners: Evidence remains lacking for effects on human gut function.

The importance of nutrient induced gut-brain signalling in the regulation of human food intake has become an increasing focus of research. Much of the caloric excess consumed comes from dietary sugars, but our knowledge about the mechanisms mediating the physiological and appetitive effects of sweet tastants in the human gut and gut-brain axis is far from complete. The comparative effects of natural sugars vs low calorie sweeteners are also poorly understood. Research in animal and cellular models has suggested a key functional role in gut endocrine cells for the sweet taste receptors previously well described in oral taste. However human studies to date have very consistently failed to show that activation of the sweet taste receptor by low calorie sweeteners placed in the human gut fails to replicate any of the effects on gastric motility, gut hormones or appetitive responses evoked by caloric sugars.

Low-calorie sweetener use and energy balance: Results from experimental studies in animals, and large-scale prospective studies in humans.

For more than a decade, pioneering animal studies conducted by investigators at Purdue University have provided evidence to support a central thesis: that the uncoupling of sweet taste and caloric intake by low-calorie sweeteners (LCS) can disrupt an animal's ability to predict the metabolic consequences of sweet taste, and thereby impair the animal's ability to respond appropriately to sweet-tasting foods. These investigators' work has been replicated and extended internationally. There now exists a body of evidence, from a number of investigators, that animals chronically exposed to any of a range of LCSs - including saccharin, sucralose, acesulfame potassium, aspartame, or the combination of erythritol+aspartame - have exhibited one or more of the following conditions: increased food consumption, lower post-prandial thermogenesis, increased weight gain, greater percent body fat, decreased GLP-1 release during glucose tolerance testing, and significantly greater fasting glucose, glucose area under the curve during glucose tolerance testing, and hyperinsulinemia, compared with animals exposed to plain water or - in many cases - even to calorically-sweetened foods or liquids. Adverse impacts of LCS have appeared diminished in animals on dietary restriction, but were pronounced among males, animals genetically predisposed to obesity, and animals with diet-induced obesity. Impacts have been especially striking in animals on high-energy diets: diets high in fats and sugars, and diets which resemble a highly-processed 'Western' diet, including trans-fatty acids and monosodium glutamate. These studies have offered both support for, and biologically plausible mechanisms to explain, the results from a series of large-scale, long-term prospective observational studies conducted in humans, in which longitudinal increases in weight, abdominal adiposity, and incidence of overweight and obesity have been observed among study participants who reported using diet sodas and other LCS-sweetened beverages daily or more often at baseline. Furthermore, frequent use of diet beverages has been associated prospectively with increased long-term risk and/or hazard of a number of cardiometabolic conditions usually considered to be among the sequelae of obesity: hypertension, metabolic syndrome, diabetes, depression, kidney dysfunction, heart attack, stroke, and even cardiovascular and total mortality. Reverse causality does not appear to explain fully the increased risk observed across all of these studies, the majority of which have included key potential confounders as covariates. These have included body mass index or waist circumference at baseline; total caloric intake and specific macronutrient intake; physical activity; smoking; demographic and other relevant risk factors; and/or family history of disease. Whether non-LCS ingredients in diet beverages might have independently increased the weight gain and/or cardiometabolic risk observed among frequent consumers of LCS-sweetened beverages deserves further exploration. In the meantime, however, there is a striking congruence between results from animal research and a number of large-scale, long-term observational studies in humans, in finding significantly increased weight gain, adiposity, incidence of obesity, cardiometabolic risk, and even total mortality among individuals with chronic, daily exposure to low-calorie sweeteners - and these results are troubling.

Recent studies of the effects of sugars on brain systems involved in energy balance and reward: Relevance to low calorie sweeteners.

The alarmingly high rates of overweight and obesity pose a serious global health threat. Numerous factors can result in weight gain, one of which is excess consumption of caloric sweeteners. In an effort to aid weight loss efforts, many people have switched from caloric sweeteners to low calorie sweeteners, which provide sweet taste without the accompanying calories. In this review, we present an overview of the animal literature produced in the last 5years highlighting the effects of sugar consumption on neural pathways involved in energy balance regulation and reward processing. We also examine the latest evidence that is beginning to elucidate the effects of low calorie sweeteners on these neural pathways, as well as how homeostatic and hedonic systems interact in response to, or to influence, sugar consumption.

Reshaping the gut microbiota: Impact of low calorie sweeteners and the link to insulin resistance?

Disruption in the gut microbiota is now recognized as an active contributor towards the development of obesity and insulin resistance. This review considers one class of dietary additives known to influence the gut microbiota that may predispose susceptible individuals to insulin resistance - the regular, long-term consumption of low-dose, low calorie sweeteners. While the data are controversial, mounting evidence suggests that low calorie sweeteners should not be dismissed as inert in the gut environment. Sucralose, aspartame and saccharin, all widely used to reduce energy content in foods and beverages to promote satiety and encourage weight loss, have been shown to disrupt the balance and diversity of gut microbiota. Fecal transplant experiments, wherein microbiota from low calorie sweetener consuming hosts are transferred into germ-free mice, show that this disruption is transferable and results in impaired glucose tolerance, a well-known risk factor towards the development of a number of metabolic disease states. As our understanding of the importance of the gut microbiota in metabolic health continues to grow, it will be increasingly important to consider the impact of all dietary components, including low calorie sweeteners, on gut microbiota and metabolic health.

Low calorie sweeteners and gut microbiota.

Studies dating back to 1980s, using bacterial cultures, have reported associations between low calorie sweeteners (LCS) and alterations in bacterial composition, raising the potential that LCS might exert effects on the host via interactions with gut microbiota. However, the results of a few recent studies carried out in this area have produced controversies. There is evidence that human fecal samples, used in most human microbiome studies, may provide a poor representation of microbial contents of the proximal intestine. Furthermore, fecal short chain fatty acid levels do not exemplify the amount of short chain fatty acids produced in the intestine. Short chain fatty acids are largely absorbed in the intestine by a tightly regulated mechanism. Here we present an exemplar study showing that the determination of the molecular mechanism(s) underlying the precise mode of action of a LCS on gut microbiota allows for rational and scientifically-based recommendations.

Functional roles of low calorie sweeteners on gut function.

This short review summarizes the effects of low calorie sweeteners (fructose, non-nutritive low calorie sweeteners) on gut functions focusing on the gut sweet taste receptor system. The effects of these molecules on secretion of gut peptides associated with glycemic homeostasis and appetite regulation is reviewed as well as effects on gastric emptying and glucose absorption.

Developing a carob-based milk beverage using different varieties of carob pods and two roasting treatments and assessing their effect on quality characteristics.

BACKGROUND: This work aimed at formulating a carob-based milk beverage and assessing its chemical and sensory properties. Six varieties of carob pods, each processed into roasted and unroasted powders, were used to develop 12 prototypes of the beverage. Chemical and physico-chemical analyses (moisture, ash, fibre, protein, sugars, total-phenolics, total-antioxidants, water activity and colour) and sensory tests were conducted. RESULTS: The variety of carob pod had a significant effect on all chemical variables in carob powders (P < 0.01), except for sugars, and when incorporated in the beverage, on moisture, total phenolics, total antioxidant activity and colour parameters (L, a, b; P-values < 0.001). Roasting treatment significantly increased fibre, total phenolics, total antioxidant activity (P-values < 0.001), fructose, glucose (P-values < 0.05), and a-value levels (P < 0.01), significantly lowered moisture (P < 0.05), water activity, L- and b-values (P-values < 0.001) in carob powders; and significantly increased the beverage's total phenolics, a-value (P-values < 0.001) and total antioxidant activity (P < 0.01). Roasting treatment significantly increased the beverage's acceptability ratings. Beverages formulated with roasted carob powder had higher ratings for level of residue, colour, caramel odour, mocha odour and flavour, roasted coffee odour and flavour, viscosity mouthfeel and bitter aftertaste. Principal component analysis was conducted; PC1 and PC2 separated attributes according to roasting treatment and variety of carob pods, respectively. CONCLUSION: The use of Akkari roasted and Baladi Ikleem el Kharoob roasted to formulate a carob-based milk beverage is recommended. © 2015 Society of Chemical Industry.

Steviol glycoside safety: are highly purified steviol glycoside sweeteners food allergens?

Steviol glycoside sweeteners are extracted from the plant Stevia rebaudiana (Bertoni), a member of the Asteraceae (Compositae) family. Many plants from this family can induce hypersensitivity reactions via multiple routes of exposure (e.g., ragweed, goldenrod, chrysanthemum, echinacea, chamomile, lettuce, sunflower and chicory). Based on this common taxonomy, some popular media reports and resources have issued food warnings alleging the potential for stevia allergy. To determine if such allergy warnings are warranted on stevia-based sweeteners, a comprehensive literature search was conducted to identify all available data related to allergic responses following the consumption of stevia extracts or highly purified steviol glycosides. Hypersensitivity reactions to stevia in any form are rare. The few cases documented in the peer-reviewed literature were reported prior to the introduction of high-purity products to the market in 2008 when many global regulatory authorities began to affirm the safety of steviol glycosides. Neither stevia manufacturers nor food allergy networks have reported significant numbers of any adverse events related to ingestion of stevia-based sweeteners, and there have been no reports of stevia-related allergy in the literature since 2008. Therefore, there is little substantiated scientific evidence to support warning statements to consumers about allergy to highly purified stevia extracts.

Effect of D-psicose used as sucrose replacer on the characteristics of meringue.

Excessive intake of sugar-rich foods leads to metabolic syndrome. D-Psicose (Psi) not commonly found in nature, is noncalorie sweetener with a suppressive effect on the blood glucose level. Thus, Psi has the potential to be utilized as a sucrose (Suc) replacer in sugar-rich foods, including meringue-based confectionery (MBC). In this study, we investigated the effect of Psi on the physical and chemical properties of meringue. Meringue was made by whipping egg white and Suc (at a weight ratio of 1:1) and baking at 93 °C for 2 h. Thirty percent of the total weight of Suc was replaced with D-ketohexoses such as Psi, D-fructose, D-tagatose, and D-sorbose. The meringues containing D-ketohexoses had higher specific volume than the meringue not containing D-ketohexoses (Ct-meringue). Baking of meringue caused differences between Psi and the other D-ketohexose meringues. Meringue containing Psi (P30-meringue) had the highest breaking stress (7.00 × 10(5) N/m(2)) and breaking strain (4.40%), resulting in the crunchiest texture. In addition, P30-meringue also had the highest antioxidant activity (491.84 µM TE/mg-meringue determined by ABTS method) and was the brownest due to a Maillard reaction occurring during baking. The replacement of Suc with Psi improved the characteristics of baked meringue. Thus, Psi was found to be useful in modifying the physical and chemical properties of MBC.

Plasma Pharmacokinetics and Routes of Excretion of [14C]-Labeled Arruva, a High-Potency Sweetener, Following Oral Administration to Beagle Dogs.

[14C]-Labeled arruva [sodium/potassium (2R,4R)-2-amino-4-carboxy-4-hydroxy-5-(3-indolyl) pentanoate] was administered as a single gavage dose (10 mg/kg bw) to male and female Beagle dogs and 1 bile duct-cannulated male. The mean peak arruva plasma concentration equivalent of 1.2 µg/g occurred at first sampling time point of 1 hour postdosing. The mean area under the concentration versus time curve from 0 hour postdosing to the last time point was approximately 20 µg·h/g and the mean terminal plasma elimination half-life ranged from 15 hours in females to 21 hours in males. Over 168 hours postdosing, 35% to 50% of the administered arruva was eliminated in the urine with 44% to 53% eliminated in feces; 1.3% of the administered dose was recovered in bile. Arruva and its derivatives were identified using tandem mass spectrometry, and the relative percentage of each substance was quantified via radio high-performance liquid chromatography. Over a 168-hour collection period, combined urine and feces extract data from the 6 noncannulated dogs showed that approximately 91% of the dose was excreted as unchanged parent arruva (41% in urine and 50% in feces). In the cannulated male, 95.3% was excreted as unchanged parent arruva; 50.2% in urine, 43.9% in feces, and 1.3% in bile. Lactone and lactam derivatives of arruva and 1 unidentified substance were detected in urine only during the first 24 hours postdosing with the greatest amounts detected during the first 6 hours of collection; up to 1% of lactone or lactam derivatives were detected in bile samples. Plasma pharmacokinetics data indicated rapid absorption of arruva with the majority of radioactivity located in the feces collected in the first 48 hours.

Structure and biological properties of lentztrehalose: a novel trehalose analog.

A new trehalose analog, lentztrehalose [4-O-(2,3-dihydroxy-3-methylbutyl)trehalose], was isolated from an actinomycete Lentzea sp. Lentztrehalose is only weakly hydrolyzed by the trehalose-hydrolyzing enzyme, trehalase, so can be regarded as an enzyme-stable analog of trehalose. Although lentztrehalose does not show apparent toxicity to mammalian cells and microbes, it has antitumor activity in mice bearing S-180 sarcoma and Ehrlich carcinoma cells. In ovariectomized mice, lentztrehalose displayed a bone reinforcement effect in the femur that was superior to trehalose and induced non-morbid suppression of weight gain comparable with trehalose. These results indicate that enzyme-stable analogs of trehalose, such as lentztrehalose, may be more beneficial for human health and thus have potential as substitutes for trehalose as a sweetener.

Administration of saccharin to neonatal mice influences body composition of adult males and reduces body weight of females.

Nutritional or pharmacological perturbations during perinatal growth can cause persistent effects on the function of white adipose tissue, altering susceptibility to obesity later in life. Previous studies have established that saccharin, a nonnutritive sweetener, inhibits lipolysis in mature adipocytes and stimulates adipogenesis. Thus, the current study tested whether neonatal exposure to saccharin via maternal lactation increased susceptibility of mice to diet-induced obesity. Saccharin decreased body weight of female mice beginning postnatal week 3. Decreased liver weights on week 14 corroborated this diminished body weight. Initially, saccharin also reduced male mouse body weight. By week 5, weights transiently rebounded above controls, and by week 14, male body weights did not differ. Body composition analysis revealed that saccharin increased lean and decreased fat mass of male mice, the latter due to decreased adipocyte size and epididymal, perirenal, and sc adipose weights. A mild improvement in glucose tolerance without a change in insulin sensitivity or secretion aligned with this leaner phenotype. Interestingly, microcomputed tomography analysis indicated that saccharin also increased cortical and trabecular bone mass of male mice and modified cortical bone alone in female mice. A modest increase in circulating testosterone may contribute to the leaner phenotype in male mice. Accordingly, the current study established a developmental period in which saccharin at high concentrations reduces adiposity and increases lean and bone mass in male mice while decreasing generalized growth in female mice.