Synergism among ternary mixtures of fourteen sweeteners

The purpose of the present study was to determine the degree of synergism of sweet taste among ternary mixtures of 14 sweeteners. A trained panel evaluated ternary mixtures of 14 sweeteners varying in chemical structure and type. The ternary mixtures that were tested were limited to those in which the compounds comprising the mixture were synergistic in binary combinations, according to an earlier study. All sweeteners in the ternary mixtures were isointense with 2% sucrose, according to a previously developed formulae. Each self-mixture was also tested (e.g. 2% sucrose + 2% sucrose + 2% sucrose). The triad with the highest mean sweetness intensity rating was alitame-neohesperidin dihydrochalcone-rebaudioside-A (10.8). This represents an increase of 99.4% when compared with the average of the self-mixtures. While this is greater than the maximum of 74% increase found for binary mixtures, more dyadic combinations of sweeteners tested previously exhibited synergism than ternary combinations tested here. However, most ternary mixtures were synergistic (significantly greater than the average of the three self-mixtures) to some degree.     

Selective inhibition of sweetness by the sodium salt of +/-2-(4-methoxyphenoxy)propanoic acid.

The purpose of this study was to determine the degree to which the sodium salt of +/-2-(4-methoxyphenoxy)propanoic acid (Na-PMP) reduced sweet intensity ratings of 15 sweeteners in mixtures. Na-PMP has been approved for use in confectionary/frostings, soft candy and snack products in the USA at concentrations up to 150 p.p.m. A trained panel evaluated the effect of Na-PMP on the intensity of the following 15 sweeteners: three sugars (fructose, glucose, sucrose), three terpenoid glycosides (monoammonium glycyrrhizinate, rebaudioside-A, stevioside), two dipeptide derivatives (alitame, aspartame), two N-sulfonylamides (acesulfame-K, sodium saccharin), two polyhydric alcohols (mannitol, sorbitol), 1 dihydrochalcone (neohesperidin dihydrochalcone), one protein (thaumatin) and one sulfamate (sodium cyclamate). Sweeteners were tested at concentrations isosweet with 2.5, 5, 7.5 and 10% sucrose in mixtures with two levels of Na-PMP: 250 and 500 p.p.m. In addition, the 15 sweeteners were tested either immediately or 30 s after a pre-rinse with 500 p.p.m. Na-PMP. In mixtures, Na-PMP at both the 250 and 500 p.p.m. levels significantly blocked sweetness intensity for 12 of the 15 sweeteners. However, when Na-PMP was mixed with three of the 15 sweeteners (monoammonium glycyrrhizinate, neohesperidin dihydrochalcone and thaumatin), there was little reduction in sweetness intensity. Pre-rinsing with Na-PMP both inhibited and enhanced sweetness with the greatest enhancements found for monoammonium glycyrrhizinate, neohesperidin dihydrochalcone and thaumatin, which were not suppressed by Na-PMP in mixtures. The mixture data suggest that Na-PMP is a selective competitive inhibitor of sweet taste. The finding that pre-treatment can produce enhancement may be due to sensitization of sweetener receptors by Na-PMP.     

POTENCY OF SWEETNESS OF ASPARTAME, D-TRYPTOPHAN AND THAUMATIN EVALUATED BY SINGLE VALUE AND TIME-INTENSITY MEASUREMENTS

Perceived sweetness of sucrose, aspartame, D-tryptophan and thaumatin in a sour, citric acid background was analyzed in terms of the potency of these compounds relative to sucrose-water combinations. Potencies of the sweeteners were determined from (1) maximum intensity using single value and time-intensity (T-I) measurements and (2) average intensity calculated as the ratio of area under the T-I curve and total perceived time. Stevens' law was applied to sweet responses, either in static or dynamic conditions. It was found that the exponent of the concentration-response function reflected the relative capacity of a compound to sweeten a given food and stressed differences of potency among sweeteners. Aspartame, D-tryptophan and thaumatin exhibited a decrease in sweetness potency relative to sucrose as sweetness increased from 10 to 100% of the full scale of response. Across the entire sweetness range, thaumatin showed the greatest potency but its long persistence time led to differentiate this intense sweetener from the other sweeteners evaluated.     

Quantitative and qualitative evaluation of high-intensity sweeteners and sweetener mixtures

High intensity sweeteners were evaluated for sweetness and bitternessintensity using time-intensity scaling. Mean intensities of50:50 mixtures as well as the single sweeteners were used tocompute predicted scores which were compared to the observedscores as a means of evaluating additivity in the mixtures.Concentration-dependent effects of subadditivity, additivityand hyperadditivity were observed within some sweetener pairs,but these did not follow any consistent pattern across sweeteners.Synergy, a special case of hyperadditivity evaluated by comparingpredicted to observed scores, was seen in mixtures of aspartameand acesulfame-K at all concentrations. Aspartame/saccharinblends were synergistic only at the lowest concentration tested,despite the structural similarity between acesulfame-K and saccharin.Blends of sucrose/aspartame and acesulfame-K/saccharin did notexhibit synergy. Comparisons based on ratings of initial sweetnessrather than the whole time-intensity curve, reflected previousfindings of synergy in some sweetener pairs.     

A REAPPRAISAL OF THE USE OF MULTIDIMENSIONAL SCALING TO INVESTIGATE THE SENSORY CHARACTERISTICS OF SWEETENERS

Previous investigations of the sensory characteristics of sweeteners using a multidimensional scaling (MDS) approach, have involved sweeteners which were not matched for sweetness. Under such circumstances, part of the estimated distance between sweeteners is attributable to sweetness differences. This detracts from the value of the consequent MDS space, when the main objective is usually to investigate sensory characteristics other than sweetness. In this study, the MDS approach was applied to sweetener solutions which were matched for sweetness with 5% sucrose. The direction of any residual sweetness differences was identified by including 1,3,5 and 7% sucrose solutions, all matched to equal viscosity, in the study. From the resulting three dimensional MDS sweetener space, it was evident that Dimension 1 was almost exclusively due to sweetness differences whereas Dimensions 2 and 3 were devoid of influence from sweetness and hence represent the sweeteners with respect to their other sensory characteristics.     

REGIONAL VARIATION IN SWEET SUPPRESSION

Differences in the sweet‐blocking efficacy of 2‐(4‐methoxyphenoxy) propanoic acid (PMP) for different sweeteners (sucrose and aspartame) and for various exposure areas of the mouth were found. Twenty participants rated sweetener solutions with and without PMP for sweetness, sourness, saltiness, bitterness and umami for stimulation of anterior tongue, posterior tongue and whole‐mouth areas. For sweetness ratings, suppression was significant for all stimulation areas. In the presence of PMP, stimulation of the posterior tongue yielded significantly higher sweetness ratings than stimulation of the anterior tongue for aspartame but not for sucrose. Sourness and bitterness ratings were significantly higher for anterior tongue than posterior tongue stimulations for aspartame but not for sucrose. The increases in sourness ratings in the presence of PMP were likely because of the sour taste PMP has at the concentration used. Results imply a difference between the front and the back of the tongue in the mechanisms involved in the perception of sweetness.     

Identification of novel sweet protein for nutritional applications

The prevalence of obesity and diabetes has increased exponentially in recent years around the globe, especially in India. Sweet proteins have the potential to substitute the sugars, by acting as natural, good and low calorie sweeteners. They also do not trigger a demand for insulin in diabetic patients unlike sucrose. In humans, the sweet taste perception is mainly due to taste-specific G protein-coupled heterodimeric receptors T1R2-T1R3. These receptors recognize diverse natural and synthetic sweeteners such as monelin, brazzein, thaumatin, curculin, mabinlin, miraculin and pentadin. Structural modeling of new sweetener proteins will be a great leap in further advancement of knowledge and their utility as sweeteners. We have explored the fingerprints of sweetness by studying the aminoacid composition and structure properties of the above proteins. The structural analysis of monellin revealed that the individual A or B chains of monellin are not contributing to its sweetness. However, the native conformation and ionic interaction between AspB7 of monellin with active site of T1R2-T1R3 receptor, along with hydrogen bonding stability of IleB6 and IleB8 are responsible for the sweet taste. Based on structural similarity search, we found a new hypothetical protein from Shewanella loihica, which has the presence of Asp(32) with adjacent isoleucine residues. Further, we examined the lead protein by two-step docking for the study of interaction of functionally conserved residues with receptors. The identified protein showed similar ionic and hydrophobic interactions with monelin. This gives a promising opportunity to explore this protein for potential health application in the low calorie sweetener industry viz., soft drinks, snacks, food, chocolate industries etc.     

Odor-taste interactions: effects of attentional strategies during exposure

Through repeated pairings with a tastant such as sucrose, odors are able to take on the tastant's qualities, e.g. by becoming more sweet smelling. When such odors are subsequently experienced with a sweet tastant in solution, the mixture is often given a higher sweetness rating than the tastant alone. Odor-induced taste enhancement appears to be sensitive to whether an odor-taste combination is viewed analytically as a set of discrete qualities, or synthetically as a flavor. The present research attempted to determine if adoption of these different perceptual approaches during co-exposure with sucrose would influence the extent to which an odor would become sweet smelling and subsequently enhance sweetness intensity. In Experiment 1, subjects received multiple exposures to mixtures of sucrose with low sweetness, low familiarity odors or, as a control, the odors and sucrose solutions separately. Two groups that received mixtures made intensity ratings that promoted either synthesis or analysis of the individual elements in the mixtures. The odors became sweeter smelling irrespective of group. Only adopting a synthetic strategy produced odors that enhanced sweetness in solution. However, these effects were also shown with a 'non-exposed' control odor. This could be accounted for if the single co-exposure with sucrose that all odors received in the pre-test was able to produce sweeter odors. A second experiment confirmed this prediction. Thus, while even a single co-exposure with sucrose is sufficient to produce a sweeter odor, the adoption of a synthetic perceptual strategy during the co-exposure is necessary to produce an odor that will enhance sweetness. These data are consistent with associative leaning accounts of how odors take on taste qualities and also support the interpretation that these effects reflect the central integration of odors and tastes into flavors.     

The Effect of Sweeteners on Perceived Viscosity

Two different sweeteners, sucrose and aspartame, were matchedin perceived sweetness intensity. These solutions were thickenedwith carboxymethylcellulose to six different viscosity levels.Sucrose and aspartame appeared to decrease perceived viscosityof the solutions at a specific sweetener concentration, at allviscosity levels. However, in a second similar experiment withthree viscosity levels and seven sucrose concentrations no effectof sucrose concentration on perceived viscosity was found. Reasonsfor these conflicting results are discussed. No definite conclusionsabout the effect of sweeteners on perceived viscosity can asyet be drawn. Chem. Senses 20: 441–450, 1995.     

Accession of sweet stimuli to receptors. I. Absolute dominance of one molecular species in binary mixtures

Intensity/time studies of sweetness response in pure solutions of each of nine different sweet stimuli have been carried out. Both variables exhibit simple power functions of the form Intensity (S) = kscns and Persistence (P) = kpcnp. In binary mixtures of these nine stimuli a depression (or negative synergism) of both sweetness intensity and persistence is observed which is predictable from the low exponents of the power functions. Combination of both power functions allows the "effective concentration" of each stimulus in a binary mixture to be calculated from its observed intensity/time characteristics. All "effective concentrations" calculable in this way show absolute dominance of one stimulus in mixtures of two irrespective of the relative proportions of the two stimuli. It is suggested that the "effective concentrations" may reflect real concentrations of a single molecular species in the microenvironment of the receptor. Thus the accession of sweet molecules to ordered, localized concentrations at the receptor is ultimately dependent on chemical structure.     

Individual differences in perceived bitterness predict liking of sweeteners

Although recent molecular studies suggest that only one receptorand one signaling pathway are involved in the perception ofsweetness, this seems to contradict everyday experience thatpeople not only have different likes and dislikes of certainsweeteners but also perceive the sweeteners differently. Onepossible explanation is that variation in liking of sweetenersis due, in part, to variation across individuals in sensitivityto nonsweet tastes, such as bitterness, which are transducedby a variety of receptors. Fifty individuals were asked to rateintensities of several taste attributes of 10 sweeteners andto give hedonic assessments of each sweetener. Additionally,their sensitivity to 6-n-propyl-3-thiouracil (PROP) was determined.Results indicated that when matched for sweetness, the perceptionof bitterness and the sweetener compound were the 2 largestfactors contributing to overall liking of a sweetener. Sensitivityto PROP did not contribute significantly to the model.     

Effect of sweetener type and of sweetener and acid levels on temporal perception of sweetness, sourness and fruitiness

Sweetness, sourness and fruitiness of 18 orange-flavored solutions,with three levels of citric acid (0.75, 1.5, 2.25 g/1) and threeequi-sweet levels of either sucrose (80, 100, 120 g/l) or aspartame(0.6, 0.7, 0.8 g/l), were evaluated by time–intensitymethodology. At these concentrations, a larger range in sournessintensity than in sweetness was produced, resulting in greatersuppression of sweetness by increasing acid levels than of sournessby increasing sweetener levels. Although aspartame samples hada longer duration of sweetness and fruitiness, sucrose and aspartamedid not interact differently with the sourness of citric acid.Fruitiness intensity and duration was enhanced by both sweetnessand sourness, but to a greater extent by sourness. Whether thisenhancement is attributable to a cognitive association of sweetnessor sourness with fruitiness or is due to the inability of thesubjects to separate sweet and sour tastes from orally perceivedfruity flavor cannot be concluded from this study.     

Binary taste mixture interactions in prop non-tasters, medium-tasters and super-tasters

It is generally assumed that the mutual, but asymmetric, suppression of the components in binary taste mixtures is an invariant property of the human psychophysical response to such mixtures. However, taste intensities have been shown to vary as a function of individual differences in sensitivity, indexed by the perceived bitterness of 6-n-propylthiouracil (PROP). To determine if these variations in taste perception influence taste mixture interactions, groups of PROP super-, medium- and non-tasters assessed four binary taste mixtures: sweet-bitter [sucrose/quinine hydrochloride (QHCl)], sweet-sour (sucrose/citric acid), salty-bitter (NaCl/QHCl) and salty-sour (NaCl/citric acid). In each experiment, subjects received factorial combinations of four levels of each of two tastants and rated individual taste intensities and overall mixture intensity. For each taste quality, super-tasters typically gave higher ratings than either medium- or non-tasters, who tended not to differ. There were also group differences in the interactions of the mixtures' components. Super-tasters rated the overall intensity of the mixtures, most likely reflecting integration of the taste components, as greater than medium- and non-tasters, who again showed few differences. In sweet-bitter mixtures, non-tasters failed to show the suppression of sweetness intensity by the highest QHCl concentration that was evident in super- and medium-tasters. These data show that the perception of both tastes and binary taste mixture interactions varies as a function of PROP taster status, but that this may only be evident when three taster groups are clearly distinguished from one another.     

植物甜蛋白Thaumatin研究进展

甜蛋白自 2 0世纪 70年代发现以来 ,一直倍受人们关注 ,而源于自然的Thaumatin是植物甜蛋白中的一种 ,它具有低热量、高甜度、安全无毒 ,并可降解为人体所需的氨基酸等多种优点 ,是一种新型甜味剂。在物质文化生活日益丰富的今天 ,人们越来越重视饮食的科学性 ,吃饱的同时更加关注所摄入食品的品质 ,无疑具多功能的非糖类物质 Thaumatin就是人们所需求的理想食品。因此 ,Thaumatin成为热门研究领域之一也就不足为怪了。1 　植物甜蛋白研究概况迄今为止 ,人们从多种植物中发现并分离出 7种甜味蛋白 [1 ]。更确切地说 ,其中 5种( Thaumatin,…     

Riboflavin-binding protein exhibits selective sweet suppression toward protein sweeteners

Riboflavin-binding protein (RBP) is well known as a riboflavin carrier protein in chicken egg and serum. A novel function of RBP was found as a sweet-suppressing protein. RBP, purified from hen egg white, suppressed the sweetness of protein sweeteners such as thaumatin, monellin, and lysozyme, whereas it did not suppress the sweetness of low molecular weight sweeteners such as sucrose, glycine, D-phenylalanine, saccharin, cyclamate, aspartame, and stevioside. Therefore, the sweet-suppressing activity of RBP was apparently selective to protein sweeteners. The sweet suppression by RBP was independent of binding of riboflavin with its molecule. Yolk RBP, with minor structural differences compared with egg white RBP, also elicited a weaker sweet suppression. However, other commercially available proteins including ovalbumin, ovomucoid, beta-lactogloblin, myoglobin, and albumin did not substantially alter the sweetness of protein sweeteners. Because a prerinse with RBP reduced the subsequent sweetness of protein sweeteners, whereas the enzymatic activity of lysozyme and the elution profile of lysozyme on gel permeation chromatography were not affected by RBP, it is suggested that the sweet suppression is caused by an interaction of RBP with a sweet taste receptor rather than with the protein sweeteners themselves. The selectivity in the sweet suppression by RBP is consistent with the existence of multiple interaction sites within a single sweet taste receptor.     

Intensity/time relationships in sweetness: evidence for a queue hypothesis in taste chemoreception

Details of reaction time, total persistence time and time ofprotracted maximum intensity of sweetness in relation to concentrationof sucrose and thaumatin are presented. Reaction time approachesa constant value at an early concentration for both sweetenersand maximum times for maximum intensity occur at lower concentrationsthan total maximum persistence. These observations seem bestexplained by a two-stage model of taste chemoreception, thefirst consisting of an orderly queue of stimulus molecules approachingthe ionophor and the second being the depolarisation at theionophor itself. The orderly queue model is capable of explainingall the temporal phenomena of sweet taste including the plateauof maximum time at maximum intensity which is observed at allconcentrations. It also offers a novel view of the way in whichintense sweeteners such as thaumatin may achieve their effectsand broadens the scope for taste modifier research in the future.     

Allelic variation of the Tas1r3 taste receptor gene selectively affects taste responses to sweeteners: evidence from 129.B6-Tas1r3 congenic mice.

The Tas1r3 gene encodes the T1R3 receptor protein, which is involved in sweet taste transduction. To characterize ligand specificity of the T1R3 receptor and the genetic architecture of sweet taste responsiveness, we analyzed taste responses of 129.B6-Tas1r3 congenic mice to a variety of chemically diverse sweeteners and glucose polymers with three different measures: consumption in 48-h two-bottle preference tests, initial licking responses, and responses of the chorda tympani nerve. The results were generally consistent across the three measures. Allelic variation of the Tas1r3 gene influenced taste responsiveness to nonnutritive sweeteners (saccharin, acesulfame-K, sucralose, SC-45647), sugars (sucrose, maltose, glucose, fructose), sugar alcohols (erythritol, sorbitol), and some amino acids (D-tryptophan, D-phenylalanine, L-proline). Tas1r3 genotype did not affect taste responses to several sweet-tasting amino acids (L-glutamine, L-threonine, L-alanine, glycine), glucose polymers (Polycose, maltooligosaccharide), and nonsweet NaCl, HCl, quinine, monosodium glutamate, and inosine 5'-monophosphate. Thus Tas1r3 polymorphisms affect taste responses to many nutritive and nonnutritive sweeteners (all of which must interact with a taste receptor involving T1R3), but not to all carbohydrates and amino acids. In addition, we found that the genetic architecture of sweet taste responsiveness changes depending on the measure of taste response and the intensity of the sweet taste stimulus. Variation in the T1R3 receptor influenced peripheral taste responsiveness over a wide range of sweetener concentrations, but behavioral responses to higher concentrations of some sweeteners increasingly depended on mechanisms that could override input from the peripheral taste system.     

Expression of the sweet protein brazzein in maize for production of a new commercial sweetener

The availability of foods low in sugar content yet high in flavour is critically important to millions of individuals conscious of carbohydrate intake for diabetic or dietetic purposes. Brazzein is a sweet protein occurring naturally in a tropical plant that is impractical to produce economically on a large scale, thus limiting its availability for food products. We report here the use of a maize expression system for the production of this naturally sweet protein. High expression of brazzein was obtained, with accumulation of up to 4% total soluble protein in maize seed. Purified corn brazzein possessed a sweetness intensity of up to 1200 times that of sucrose on a per weight basis. In addition, application tests demonstrated that brazzein-containing maize germ flour could be used directly in food applications, providing product sweetness. These results demonstrate that high-intensity sweet protein engineered into food products can give sweetener attributes useful in the food industry.     

THE TASTES OF ARTIFICIAL SWEETENERS AND THEIR MIXTURES

0s scaled the taste intensity (bitterness, sweetness) of artificialsweeteners, mixtures of artificial sweeteners, and glucose.Sweetness of glucose conformed to a power function, whereasneither sweetness of artificial sweeteners nor their bitternessdid. The total taste intensity of mixtures was often lower thanthe taste intensities of the components, suggesting suppression,although in many instances the suppressive effects disappearedat high concentrations.     

Taste responses to sweet stimuli in alpha-gustducin knockout and wild-type mice

The importance of alpha-gustducin in sweet taste transduction is based on data obtained with sucrose and the artificial sweetener SC45647. Here we studied the role of alpha-gustducin in sweet taste. We compared the behavioral and electrophysiological responses of alpha-gustducin knockout (KO) and wild-type (WT) mice to 11 different sweeteners, representing carbohydrates, artificial sweeteners, and sweet amino acids. In behavioral experiments, over 48-h preference ratios were measured in two-bottle preference tests. In electrophysiological experiments, integrated responses of chorda tympani (CT) and glossopharyngeal (NG) nerves were recorded. We found that preference ratios of the KO mice were significantly lower than those of WT for acesulfame-K, dulcin, fructose, NC00174, D-phenylalanine, L-proline, D-tryptophan, saccharin, SC45647, sucrose, but not neotame. The nerve responses to all sweeteners, except neotame, were smaller in the KO mice than in the WT mice. The differences between the responses in WT and KO mice were more pronounced in the CT than in the NG. These data indicate that alpha-gustducin participates in the transduction of the sweet taste in general.