Taste of common prebiotic oligosaccharides: Impact of molecular structure.

Prebiotic oligosaccharides are naturally occurring non-digestible carbohydrates with demonstrated health benefits. They are also a chemically diverse class of nutrients, offering an opportunity to investigate the impact of molecular structure on oligosaccharide taste perception. Accordingly, a relevant question is whether these compounds are detected by the human gustatory system, and if so, whether they elicit sweet or 'starchy' taste. Here, in three psychophysical experiments, we investigated the taste perception of three commercially popular prebiotics [fructooligosaccharides (FOS), galactooligosaccharides (GOS), xylooligosaccharides (XOS)] in highly pure form. Each of these classes of prebiotics differs in the type of glycosyl residue, and position and type of bond between those residues. In experiments I and II, participants were asked to discriminate a total of 9 stimuli [FOS, GOS, XOS; degree of polymerization (DP) of 2, 3, 4] prepared at 75 mM in the presence and absence of lactisole, a sweet receptor antagonist. We found that all nine compounds were detectable (p<0.05). We also found that GOS and XOS DP 4 were discriminable even with lactisole, suggesting that their detection is not via the canonical sweet receptor. Accordingly, in experiment III, the taste of GOS and XOS DP 4 were directly compared with that of MOS (maltooligosaccharides) DP 4-6, which has been reported to elicit 'starchy' taste. We found that GOS and MOS were perceived similarly although narrowly discriminable, while XOS was easily discriminable from both GOS or MOS. The current findings suggest that molecular structure of oligosaccharides impacts their taste perception in human.

Evaluation of Functional Spray Coatings for Mitigating the Uptake of Volatile Phenols by Pinot Noir Wine Grapes via Blocking, Absorption, and/or Adsorption.

Spray coatings have shown promising potential in preventing the uptake of smoke phenols from wildfires into wine grapes. Three cellulose nanofiber-based coatings with low methoxyl pectin or varying concentrations of chitosan were made into films and their potential for blocking, absorption, or adsorption of phenols (guaiacol, m-cresol, and syringol) was evaluated using a custom-built smoke diffusion box. The coatings were also applied to Pinot noir grapes in a vineyard. GC-MS analysis for smoke phenols from headspace gases of diffusion study and extractions of films indicated that chitosan-based films can block guaiacol and syringol, and all films are able to capture m-cresol. The type of coating and application time in a vineyard did not affect (P < 0.05) physicochemical properties, size, and weight of the berries, whereas chitosan-based coatings resulted in a higher anthocyanin content of berries. This study provided new information about the key mechanisms (i.e., blocking phenols) of coatings to mitigate smoke phenol uptake in wine grapes.

Taste perception of oligosaccharides derived from pullulan.

Recent studies indicate that humans can taste starch hydrolysis products (i.e. maltooligosaccharides; MOS). However, the structural specificity of oligosaccharides that elicit such perception is not known. This study investigated taste perception of pullulan-derived oligosaccharides (PDOS) that are structurally similar to MOS, but differ in that every third glycosidic linkage in PDOS is α-1,6, rather than α-1,4. Three food-grade PDOS stimuli were produced by limited-enzyme hydrolysis of pullulan. The resulting products were stimuli with degree of polymerization (DP) of 3, 6, and 9. Subjects discriminated all 3 stimuli from blanks at a significant level (P < 0.00001) in the absence of lactisole, a sweet taste inhibitor. In the presence of lactisole, the subjects could not detect DP 3 at a significant level (P > 0.05), but were able to detect DP 6 and 9 (P < 0.005), although the degree of detectability dropped significantly (P < 0.05). In a follow-up qualitative study, subjects made the target stimuli and glucose into 2 groups (glucose/DP 3 vs. DP 6/DP 9) and characterized both groups as mostly "sweet" with having different sweetness intensity. With lactisole, they described glucose and DP 3 as "taste like blank" (lactisole water) and found it challenging to describe DP 6 and 9 stimuli due to their subtle nature. These results suggest that taste perception of PDOS primarily depends on the sweet taste receptor, although they may elicit other sensory attributes; this is strikingly different from the reported taste of MOS. The potential impact of structural configuration on taste perception is further discussed.

Taste Detection of Maltooligosaccharides with Varying Degrees of Polymerization.

Previous studies have shown that humans can taste maltooligosaccharides [MOS; degree of polymerization (DP) of 3-20] but not maltopolysaccharides (MPS; DP of >20) and that their taste detection is independent of the canonical sweet taste receptor. The objectives of this study were to determine the DP ranges of target stimuli that are tasted and further to investigate the impact of DP on taste detectability. To achieve this goal, we prepared three food-grade MOS samples with narrow DP ranges using flash chromatography: low (4-6), medium (7-12), and high (14-21) DP samples. Following sample preparation, we asked subjects to discriminate the MOS stimuli from blanks after the stimuli were swabbed on the tip of tongue. All stimuli were initially presented at 75 mM. Acarbose, an α-glucosidase inhibitor, was added to all stimuli, including blanks, to prevent oral hydrolysis of MOS. After determining that all three MOS samples were detected at a significant degree, we conducted follow-up studies to explore whether the detection of these samples differed at a range of concentrations (18-56 mM). The results showed that detection rates of medium- and high-DP MOS varied in a concentration-dependent manner (p < 0.05). In contrast, low-DP MOS showed a consistent detection rate across concentrations tested. These results demonstrate that humans can taste MOS stimuli of all chain lengths and that relative taste detection rates are generally similar across MOS with varying chain lengths.

Impact of functional spray coatings on smoke volatile phenol compounds and Pinot noir grape growth.

The frequency and intensity of wildfires have been increasing over the last 50 years and negatively impacted the wine industry. Previous methods of smoke mitigation during grape processing have shown little impact in reducing smoke taint in wines. Therefore, a novel method of using edible spray coatings for vineyard application was developed to help prevent volatile smoke phenol uptake in wine grapes. Four cellulose nanofiber-based coating suspensions incorporated with chitosan and/or ß-cyclodextrin were evaluated. Films derived from the coating suspensions were exposed to volatile phenols found in wildfire smoke (guaiacol, 4-methyl guaiacol, m-cresol, o-cresol, p-cresol, syringol, and 4-methyl syringol) and evaluated with ultraviolet-visible spectroscopy where the results indicated that the coatings could uptake smoke phenols in varying degrees. The coatings were also applied in a vineyard at three different application times during grape growth: pea-sized, pre-bunch closure, and both at pea-sized and pre-bunch closure. The results showed that the application time did not have a significant (p < 0.05) effect on berry size, weight, °Brix, pH, or titratable acidity. The type of coating, time of application and washing were found to impact the number of volatile phenols in the grapes after a smoke event. Results from this study indicated that edible coatings could help mitigate smoke uptake in wine grapes without sacrificing the growth and key composition parameters of wine grapes. PRACTICAL APPLICATION: This research provides a novel spray coating that can be applied to wine grapes in the vineyard to potentially mitigate volatile smoke compounds in wine grapes without impacting fruit growth and key compositional parameters of wine grapes, thus maintaining high quality of wines for consumers. Results from this study can also be potentially applied to other agricultural commodities to solve the issues caused by the wildfire smoke.

Chromatographic fractionation of food-grade oligosaccharides: Recognizing and avoiding sensory-relevant impurities.

Flash chromatography utilizing microcrystalline cellulose (MCC) stationary phases and aqueous ethanol mobile phases have shown promise for the production of food-grade oligosaccharides. The current work extends the scope of these systems by demonstrating their use for the production of food-grade maltooligosaccharide preparations enriched in high degree of polymerization (DP) components. Furthermore, it is shown herein that caution must be exercised when using these MCC-based chromatographic systems in order to avoid sensory-relevant contamination of the final oligosaccharide preparations. Such contamination, most notably off-taste, is shown to arise from impurities common to commercially available MCC that manifest under certain chromatographic scenarios. A mitigation strategy based on washing the stationary phase with appropriate aqueous-ethanol solutions (i.e., accounting for the entire mobile phase concentration range) prior to oligosaccharide fractionation is presented as a means by which to avoid contamination.

The Effect of p-Coumaric Acid on Browning Inhibition in Potato Polyphenol Oxidase-Catalyzed Reaction Mixtures.

There has been considerable interest in using natural polyphenol oxidase (PPO) inhibitors to control browning in fruit and vegetable products. p-Coumaric acid (pCA), a common secondary metabolite of plants, has been studied as an inhibitor of PPOs/tyrosinases from several foods (e.g., mushroom, apple, and potato). However, studies on the use of pCA for the inhibition of PPO-initiated browning in actual food systems are limited. Therefore, a study was carried out to ascertain the efficacy of using pCA to limit PPO-initiated browning in fresh potato juice. The extent of browning inhibition by pCA was shown to be reaction system-dependent. Browning in potato juice was unexpectedly enhanced by the addition of pCA. This was interpreted as pCA acting as an alternative substrate with significantly higher browning efficiency; extent of browning under this condition was higher than that observed in the native potato juice. The addition of pCA to any of the model reaction mixtures (i.e., those containing semi-purified enzymes and substrates) significantly inhibited browning. The discrepancy in pCA effects on browning inhibition in different reaction systems is postulated to be mainly due to non-enzyme and non-substrate components in potato juice that participate in the post-PPO reaction sequences, which ultimately lead to brown color formation.

Chromatographic preparation of food-grade prebiotic oligosaccharides with defined degree of polymerization.

Prebiotic oligosaccharides are of widespread interest in the food industry due to their potential health benefits. This has triggered a need for research into their sensory properties. Such research is currently limited due to the lack of available food-grade oligosaccharide preparations with specific degree of polymerization (DP). The aim of this study was to develop economical approaches for the preparation and characterization of prebiotic oligosaccharides differing with respect to composition and DP. Such preparations were prepared by chromatographic fractionation of commercially available prebiotic mixtures using microcrystalline cellulose stationary phases and aqueous ethanol mobile phases. This approach is shown to work for the preparation of food-grade fructooligosaccharides of DP 3 and 4, galactooligosaccharides of DP 3 and 4, and xylooligosaccharides of DP 2-4. Methods for the characterization of the different classes of oligosaccharides are also presented including those addressing purity, identity, total carbohydrate content, moles per unit mass, and DP.

Role of Ascorbic Acid in the Extraction and Quantification of Potato Polyphenol Oxidase Activity.

The ability to accurately measure the activity of polyphenol oxidase (PPO) in complex matrices is essential. A problem encountered when using spectrophotometric methods is interference due to ascorbic acid (AA), often used as an enzyme "protecting agent" during PPO extraction. This study focuses on the nature of AA's effect on spectrophotometric determinations of PPO activity as well as enzyme extraction. Potato extracts and semi-purified PPO were used as enzyme sources. The inactivation of PPO attributed to AA is substrate-mediated. The extent of AA-dependent inactivation of PPO in model systems varied between substrates. AA only slows mechanism-based inactivation of PPO induced by catechol, possibly owing to the prevention of quinone formation. AA minimally protects PPO activity during enzyme extraction. The problem associated with AA in PPO assay could be circumvented by using ascorbate oxidase to remove AA when catechol is the primary substrate or by using chlorogenic acid as the primary substrate.

Information Retrieval in Food Science Research II: Accounting for Relevance When Evaluating Database Performance.

The impetus for this study is the limited amount of information on performance characteristics of food science-encompassing online bibliographic databases. Database usage is an important element in modern research because a comprehensive understanding of current knowledge is essential for effective, unbiased hypothesis formulation and testing. Six databases commonly recommended by academic libraries for information retrieval in the food sciences (Academic Search Premier [ASP], Agricultural Online Access [AGRICOLA], CAB Direct, Food Science and Technology Abstracts [FSTA], PubMed, and Web of Science [WoS]) were compared in a case study based on the research topic "in vitro bile acid binding properties of dietary lignin." A complex string of search terms was used for citation retrieval, and predetermined inclusion/exclusion criteria were used to determine the relevancy of retrieved articles. Searching WoS and CAB Direct returned the greatest number of relevant articles, followed closely by FSTA, PubMed, and AGRICOLA. FSTA and AGRICOLA returned the highest ratios of relevant-to-irrelevant articles, followed closely by CAB Direct. None of the databases, when used alone, recovered all of the relevant articles identified in the study; WoS indexed the highest percentage of relevant articles identified (WoS = 10; total = 19). WoS also had the highest number of relevant articles that were unique to any one database. The thoroughness of searching the complete group of databases was tested by comparison of retrieved citations with those found in relevant review articles, revealing the need for testing overall inclusivity. PRACTICAL APPLICATION: Comprehensive online information retrieval is the most efficient means of accessing current knowledge. Awareness of current knowledge is essential for effective, unbiased decision making in private sector and academic/government-based research endeavors. Thus, online database usage is an essential element of modern food science research. This paper provides tangible examples of the performance characteristics of online bibliographic databases commonly recommended for information retrieval in the food sciences. The paper is written so as to aid the reader in making decisions with respect to database usage for the recovery of topic-relevant peer-reviewed articles germane to their area of research.

The nature of ß-cyclodextrin inhibition of potato polyphenol oxidase-catalyzed reactions.

There is general interest in strategies to control polyphenol oxidase (PPO)-initiated enzymatic browning because it is often associated with declining food quality. Cyclodextrins are cyclic glucan oligosaccharides that form inclusion complexes with a number of PPO substrates. This study focuses on the effect of ß-cyclodextrins (ßCyD) on PPO-catalyzed reactions. Potato enzyme extracts and semi-purified potato PPO served as enzyme sources. Substrates included phenolics endogenous to potatoes. Reaction time-courses were followed spectrophotometrically; rates were compared by analysis of variance. Extents of ßCyD inhibition of PPO-catalyzed reactions are shown to be substrate specific and can be quantitatively accounted for based on degrees of ßCyD substrate sequestration. There was no evidence for direct irreversible ßCyD inactivation of potato PPO. An apparent "direct PPO inactivation" by ßCyD is shown to result from a sequence of sequestration-dependent reactions that occur in commonly employed assay systems for the quantification of PPO in fruits and vegetables.

The Sweet Taste of Acarbose and Maltotriose: Relative Detection and Underlying Mechanism.

Although sweet-tasting saccharides possess similar molecular structures, their relative sweetness often varies to a considerable degree. Current understanding of saccharide structure/sweetness interrelationships is limited. Understanding how certain structural features of saccharides and/or saccharide analogs correlate to their relative sweetness can provide insight on the mechanisms underlying sweetness potency. Maltotriose is a short-chain glucose-based oligosaccharide, which we recently reported to elicit sweet taste. Acarbose, an α-glucosidase inhibitor, is a pseudo-saccharide that has an overall resemblance to a glucose-based oligosaccharide and thus may be viewed as a structural analog. During other studies, we recognized that acarbose can also elicit sweet taste. Here, we formally investigated the underlying taste detection mechanism of acarbose, while confirming our previous findings for maltotriose. We found that subjects could detect the sweet taste of acarbose and maltotriose in aqueous solutions but were not able to detect them in the presence of a sweet taste inhibitor lactisole. These findings support that both are ligands of the human sweet taste receptor, hT1R2/hT1R3. In a separate experiment, we measured the relative sweetness detection of acarbose, maltotriose, and other sweet-tasting mono- and disaccharides (glucose, fructose, maltose, and sucrose). Whereas maltotriose was found to have a similar discriminability profile to glucose and maltose, the discriminability of acarbose matched that of fructose at the concentrations tested (18, 32, and 56 mM). These findings are discussed in terms of how specific molecular features (e.g., degree of polymerization and monomer composition) may contribute to the relative sweetness of saccharides.

Information Retrieval in Food Science Research: A Bibliographic Database Analysis.

The aim of the present research was to ascertain the importance of electronic bibliographic database selection and multiple database usage during the information retrieval phase of research in the food sciences. Six commonly recommended databases were compared with respect to overall journal coverage and journal overlap. Databases were also evaluated with respect to coverage of food science-based journals and the extent of article coverage therein. A case study approach, focused on bile acid/dietary fiber interactions, was used to illustrate the ramifications of database selection/usage when dealing with specific research topics. Databases differed with respect to the breadth of disciplines covered, the total number of journals indexed, the number of food science discipline-specific journals indexed, and the number of articles included per indexed journal. All of the databases contained citations that were unique to the given database. The data resulting from the case study provide an example of the extent to which relevant information may be missed if pertinent databases are not mined. In the present case, over half of the articles retrieved on the focus research topic were unique to a single database. The combined data from this study point to the importance of thoughtful database selection and multiple database usage when comprehensively assessing knowledge in the food sciences. PRACTICAL APPLICATION: This paper provides insights into article database usage for food science-relevant information retrieval. Online information retrieval is an efficient way to assess current knowledge in any of the food science disciplines. Acquired knowledge in turn is the underpinning of effective problem solving; whether it be private sector- or academic/government-based research.

Preparation and characterization of isolated low degree of polymerization food-grade maltooligosaccharides.

Research involving human responses to the consumption of starch and its hydrolysis products would benefit from convenient sources of well defined, low cost, food-grade maltooligosaccharides (MOS). This report addresses such need by presenting an approach to obtain aforementioned MOS. A chromatography-ready MOS sample containing proportionately high amounts of low degree of polymerization (DP) MOS is initially prepared from commercially-available maltodextrins (MD) by taking advantage of the DP-dependent differential solubility of MOS in aqueous-ethanol solutions. The low DP-enriched MOS preparation is subsequently fractionated via preparative column chromatography using cellulose-based stationary phases and step-gradient aqueous-ethanol mobile phases. The resulting fractions yielded isolated food-grade MOS ranging in DP from 3 to 7. NMR spectra of isolated MOS indicated minimal amounts of branched saccharides. Typical yields from a single fractionation protocol (2 g MD starting material), including solvent partitioning through preparative chromatography, ranged from ∼40 mg for DP 4, 5, and 7 to ∼100 mg for DP 3 and 6.

Furfural and 5-hydroxymethyl-furfural degradation using recombinant manganese peroxidase.

Biomass pretreatment-derived degradation compounds, such as furfural and 5-hydroxymethyl-furfural (HMF), inhibit the growth of fermentation microorganisms that utilize biomass to produce fuels and chemicals. Here we report that recombinant manganese peroxidase (rMnP) produced from the yeast Pichia pastoris can degrade furfural and HMF making them less toxic to microorganisms. Treatment with rMnP or manganese(III) acetate reduced furfural and HMF concentrations in a dose-dependent manner. Furfural disappearance was accompanied by malonate disappearance and accumulation of four distinct degradation products. Furfural was more readily degraded by rMnP and manganese(III) acetate than HMF. Growth assays using Saccharomyces cerevisiae indicated that rMnP treatment reduced the toxicity of furfural and HMF. This work provides an avenue to use rMnP to increase the growth of fermentation microorganisms that are inhibited by toxic compounds derived from pretreatment of biomass.

Oral Digestion and Perception of Starch: Effects of Cooking, Tasting Time, and Salivary α-Amylase Activity.

Since starch is a significant part of human diet, its oral detection would be highly beneficial. This study was designed to determine whether starch or its degradation products can be tasted and what factors influence its perception. Subjects were asked 1) to taste 8% raw and cooked starch samples for 5, 15, and 35 s and rate perceived intensities of sweetness and "other" taste (i.e., other than sweet), 2) to donate saliva to obtain salivary flow rate (mg/s) and salivary α-amylase activity (per mg saliva), and 3) to fill out a carbohydrate consumption survey. Subsequently, in vitro hydrolysis of starch was performed; saliva was collected from 5 subjects with low and high amylase activities and reacted with 8% raw and cooked starch at 2, 15, and 30 s. Hydrolysis products were then quantified using a High performance liquid chromatography. The results showed cooking increased the digestibility of starch such that the amount of hydrolysis products increased with reaction time. However, cooking did not influence taste ratings, nor were they influenced by tasting time. Subjects' salivary amylase activities were associated with the efficacy of their saliva to degrade starch, in particular cooked starch, and thus the amount of maltooligosaccharide products generated. Effective α-amylase activity [i.e. α-amylase activity (per mg saliva) × salivary flow rate (mg/s)] and carbohydrate consumption score (i.e. consumption frequency × number of servings) were also independently associated with sensory taste ratings. Human perception of starch is undoubtedly complex as shown in this study; the data herein point to the potential roles of salivary α-amylase activity and carbohydrate consumption in the perception of cooked starch.

Methylene blue as a lignin surrogate in manganese peroxidase reaction systems.

Manganese peroxidase (MnP) is associated with lignin degradation and is thus relevant to lignocellulosic-utilization technologies. Technological applications require reaction mixture optimization. A surrogate substrate can facilitate this if its susceptibility to degradation is easily monitored and mirrors that of lignin. The dye methylene blue (MB) was evaluated in these respects as a surrogate substrate by testing its reactivity in reaction mixtures containing relevant redox mediators (dicarboxylic acids, fatty acids). Relative rates of MB degradation were compared to available literature reports of lignin degradation under similar conditions, and suggest that MB can be a useful lignin surrogate in MnP systems.

Human taste detection of glucose oligomers with low degree of polymerization.

Studies have reported that some animals, including humans, can taste mixtures of glucose oligomers (i.e., maltooligosaccharides, MOS) and that their detection is independent of the known T1R2/T1R3 sweet taste receptor. In an effort to understand potential mechanisms underlying the taste perception of glucose oligomers in humans, this study was designed to investigate: 1) the variability of taste sensitivity to MOS with low degree-of-polymerization (DP), and 2) the potential role of hT1R2/T1R3 in the MOS taste detection. To address these objectives, a series of food grade, narrow-DP-range MOS were first prepared (DP 3, 3-4, 5-6, and 6-7) by fractionating disperse saccharide mixtures. Subjects were then asked to discriminate these MOS stimuli as well as glucose (DP 1) and maltose (DP 2) from blanks after the stimuli were swabbed on the tongue. All stimuli were presented at 75 mM with and without a sweet taste inhibitor (lactisole). An α-glucosidase inhibitor (acarbose) was added to all test stimuli to prevent oral digestion of glucose oligomers. Results showed that all six stimuli were detected with similar discriminability in normal tasting conditions. When the sweet receptor was inhibited, DP 1, 2, and 3 were not discriminated from blanks. In contrast, three higher-DP paired MOS stimuli (DP 3-4, 5-6, and 6-7) were discriminated from blanks at a similar degree. Overall, these results support the presence of a sweet-independent taste perception mechanism that is stimulated by MOS greater than three units.

Overcoming Reductant Interference in Peroxidase-Based Assays for Hydrogen Peroxide Quantification.

A problem commonly encountered when using peroxidase-based methods for hydrogen peroxide quantification in biobased matrixes is interference due to the presence of endogenous reductants. Such assays are typically based on the generation of an oxidized reporter molecule in direct proportion to the amount of hydrogen peroxide reduced in the peroxidase-catalyzed reaction. Endogenous reductants confound such assays by reducing the oxidized reporter molecule, thus resulting in underestimates of hydrogen peroxide content. In the present work, we demonstrate how this problem can be circumvented by selectively oxidizing offending compounds by treatment with the oxidized reporter molecule prior to initiating the peroxidase reaction for hydrogen peroxide quantification. The approach is demonstrated using horseradish peroxidase, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), as the reporter molecule and a representative garlic paste as the hydrogen peroxide-containing biobased matrix. The approach is expected to be generally applicable to a wide range of peroxidase-based assays when applied to complex biobased systems.

Humans Can Taste Glucose Oligomers Independent of the hT1R2/hT1R3 Sweet Taste Receptor.

It is widely accepted that humans can taste mono- and disaccharides as sweet substances, but they cannot taste longer chain oligo- and polysaccharides. From the evolutionary standpoint, the ability to taste starch or its oligomeric hydrolysis products would be highly adaptive, given their nutritional value. Here, we report that humans can taste glucose oligomer preparations (average degree of polymerization 7 and 14) without any other sensorial cues. The same human subjects could not taste the corresponding glucose polymer preparation (average degree of polymerization 44). When the sweet taste receptor was blocked by lactisole, a known sweet inhibitor, subjects could not detect sweet substances (glucose, maltose, and sucralose), but they could still detect the glucose oligomers. This suggests that glucose oligomer detection is independent of the hT1R2/hT1R3 sweet taste receptor. Human subjects described the taste of glucose oligomers as "starchy," while they describe sugars as "sweet." The dose-response function of glucose oligomer was also found to be indistinguishable from that of glucose on a molar basis.