Isolation, identification and sensory evaluation of kokumi peptides from by-products of enzyme-modified butter.

BACKGROUND: Enzyme-modified butter is used as a common raw material to obtain a natural milk flavor. Butter protein is a by-product in butter processing that can be used as substrate to produce taste-active peptides, which can create additional value and new application opportunities, making the method more environmentally friendly. RESULTS: Putative kokumi peptides from hydrolysates of protein by-products were isolated by gel filtration chromatography and reversed-phase high-performance liquid chromatography. The isolated peptide fraction with the most pronounced kokumi taste was screened by sensory evaluation and electronic tongue analysis. Eleven peptides were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Six peptides were synthesized to verify their taste characteristics. Five synthetic peptides (FTKK, CKEVVRNANE, EELNVPG, VPNSAEER and YPVEPFTER) showed different intensity levels of kokumi taste. Of these peptides, the decapeptide CKEVVRNANE had the highest kokumi intensity. CONCLUSION: The newly identified kokumi peptides increased the kokumi taste intensity and showed some synergistic effect with umami taste. Both termini of the peptides seem to play an important role in taste characteristic. Glu residue at both termini can increase the kokumi taste intensity. This work indicated that it was feasible to produce kokumi peptides by enzymatic hydrolysis of the protein by-products of butter. © 2022 Society of Chemical Industry.

Production of wheat gluten hydrolysates with reduced antigenicity employing enzymatic hydrolysis combined with downstream unit operations.

BACKGROUND: Due to allergies or other health disorders a certain segment of the population is not able to safely consume some plant proteins, which are the main protein support in human nutrition. Coeliac disease is a prominent autoimmune disorder and requires a strict adherence to a gluten-free diet. The aim of this study was to identify suitable combinations of enzymatic hydrolysis and common unit operations in food processing (centrifugation, ultra-filtration) to produce gluten-free wheat gluten hydrolysates for food application. To analyse the hydrolysates, a simple and cheap competitive ELISA protocol was designed and validated in this study as well. RESULTS: The competitive ELISA was validated using gliadin spiked skim milk protein hydrolysates, due to the latter application of the assay. The limit of quantification was 4.19 mg kg(-1) , which allowed the identification of gluten-free (<20 mg kg(-1) ) hydrolysates. Enzymatic hydrolysis, including the type of peptidase, and the downstream processing greatly affected the antigenicity of the hydrolysates. CONCLUSION: Enzymatic hydrolysis and downstream processing operations, such as centrifugation and ultra-filtration, reduced the antigenicity of wheat gluten hydrolysates. Gluten-free hydrolysates were obtained with Flavourzyme after centrifugation (25 g L(-1) substrate) and after 1 kDa ultra-filtration (100 g L(-1) substrate). A multiple peptidase complex, such as Flavourzyme, seems to be required for the production of gluten-free hydrolysates. © 2015 Society of Chemical Industry.

Odorants Identified in Chinese Dry-Cured Ham Contribute to Salty Taste Enhancement.

Jinhua dry-cured ham (JDH) is a traditional fermented Chinese meat product. We studied the dynamic sensory and emotional profiles of JDHs obtained by five preparation methods and the corresponding release of sodium ions (Na+), potassium ions (K+), and volatile organic compounds (VOCs) during oral processing. The VOCs with salty taste enhancement abilities were screened based on the correlations of VOCs with salty flavor and concentration of Na and K ions with salty flavor. A trained sensory panel evaluated the saltiness enhancements of selected VOCs by using static and dynamic sensory methods. The results revealed that Na+, K+, and selected VOCs were mainly released during 0-10 s of the chewing process. The release of Na+ and K+ in JDH residue samples exhibited consistently decreasing trends, while in saliva, their concentrations increased. The VOCs showing a high correlation with Na+ and K+ and salty flavor have saltiness enhancement abilities in both NaCl solutions and NaCl + MSG mixtures. Odor-induced saltiness was pronounced at low salt concentrations (0.2% NaCl). The investigation demonstrated 16 VOCs exhibiting saltiness enhancement abilities, including 4 pyrazines, 5 acids, 4 sulfur-containing compounds, and 3 other compounds. The sensory evaluation suggested pyrazines and sulfur-containing compounds as good saltiness enhancers. 2-Furfuryl mercaptan significantly enhanced the salty sensation in the NaCl + MSG solutions when compared with MSG alone (p < 0.05). This research provides evidence that certain odorants identified in JDHs exhibit salty-enhancing properties, indicating their potential for salt reduction at the industrial level.

Novel Umami Peptides from Mushroom (Agaricus bisporus) and Their Umami Enhancing Effect via Virtual Screening and Molecular Simulation.

This study aimed to identify novel umami peptides in Agaricus bisporus and investigate their umami enhancing effect. We virtually screened 155 potential umami peptides from the ultrasound-assisted A. bisporus hydrolysate according to Q values, iUmami-SCM, Umami_YYDS, and Tastepeptides_DM models, and molecular docking. Five peptides (AGKNTNGSQF, DEAVARGATF, REESDFQSSF, SEETTTGVHH, and WNNDAFQSSTN) were synthesized for sensory evaluation and kinetic analysis. The result showed that the umami thresholds of the five peptides were in the range of 0.21-0.40 mmol/L. Notably, REESDFQSSF, SEETTTGVHH, and WNNDAFQSSTN had low dissociation constant (KD) values and high affinity for the T1R1-VFT receptor. The enhancing effect of the three peptides with MSG or IMP was investigated by sensory evaluation, kinetic analysis, and molecular dynamics simulations. In stable complexes, ARG_277 in T1R1 played a major role in umami peptide binding to T1R1-VFT. These results provide a theoretical basis for future screening of umami peptides and improving the umami taste of food containing mushrooms.

Differences of Typical Wuyi Rock Tea in Taste and Nonvolatiles Profile Revealed by Multisensory Analysis and LC-MS-Based Metabolomics.

Wuyi Rock tea, specifically Shuixian and Rougui, exhibits distinct sensory characteristics. In this study, we investigated the sensory and metabolite differences between Shuixian and Rougui. Quantitative description analysis revealed that Rougui exhibited higher intensity in bitter, thick, harsh, and numb tastes, while Shuixian had stronger salty and umami tastes. Nontargeted metabolomics identified 151 compounds with 66 compounds identified as key differential metabolites responsible for metabolic discrimination. Most of the catechins and flavonoids were enriched in Rougui tea, while epigallocatechin-3,3'-di-O-gallate, epigallocatechin-3,5-di-O-gallate, gallocatechin-3,5-di-O-gallate, isovitexin, and theaflavanoside I were enriched in Shuixian tea. Catechins, kaempferol, quercetin, and myricetin derivatives were positively correlated with bitter taste and numb sensation. Sour taste was positively correlated to organic acids. Amino acids potentially contributed to salty and umami tastes. These results provide further insights into the taste characteristics and the relationship between taste attributes and specific metabolites in Wuyi Rock tea.

Elucidation of the Impact of Steaming on the Key Odorants of Jinhua Dry-Cured Ham Using the Sensomics Approach.

Jinhua dry-cured ham (JDH) is a traditional fermented meat product favored by Chinese consumers. In this paper, the impact of steaming on the key odorants of JDH was investigated using the sensomics approach. Compounds with odor activity values (OAV) ≥1 were re-engineered in a triglyceride matrix to imitate the odor profiles of both raw and steamed JDHs. The aroma-active compounds were then confirmed by recombination and omission tests using triangle tests. The odor profiles of raw and steamed JDHs were obtained by quantitative descriptive analysis to compare the differences between the original and recombined models. The results showed that pentanal, hexanal, dimethyl trisulfide, (E,E)-2,4-decadienal, (E)-2-heptenal, furaneol, 3-methylbutanoic acid, 1-octen-3-one, and methional influenced the overall raw JDH odor significantly. Furaneol was first reported as a key compound that provides a caramel smell to the raw JDH. Apart from (E)-2-heptenal, dimethyl trisulfide, furaneol, 3-methylbutanoic acid, and methional, the remaining three compounds including 2-furfurylthiol, benzeneacetaldehyde, and phenylethyl alcohol showed a significant influence on the odor profile of steamed JDH. The statistical analysis of the odor profiles showed an 80.0% similarity between the recombination raw JDH and the real raw JDH, and a 76.3% similarity between the model and the real steamed JDH.

TastePeptides-EEG: An Ensemble Model for Umami Taste Evaluation Based on Electroencephalogram and Machine Learning.

In the field of food, the sensory evaluation of food still relies on the results of manual sensory evaluation, but the results of human sensory evaluation are not universal, and there is a problem of speech fraud. This work proposed an electroencephalography (EEG)-based analysis method that effectively enables the identification of umami/non-umami substances. First, the key features were extracted using percentage conversion, standardization, and significance screening, and based on these features, the top four models were selected from 19 common binary classification algorithms as submodels. Then, the support vector machine (SVM) algorithm was used to fit the outputs of these four submodels to establish TastePeptides-EEG. The validation set of the model achieved a judgment accuracy of 90.2%, and the test set achieved a judgment accuracy of 77.8%. This study discovered the frequency change of α wave in umami taste perception and found the frequency response delay phenomenon of the F/RT/C area under umami taste stimulation for the first time. The model is published at www.tastepeptides-meta.com/TastePeptides-EEG, which is convenient for relevant researchers to speed up the analysis of umami perception and provide help for the development of the next generation of brain-computer interfaces for flavor perception.

Strategies to improve meat-like properties of meat analogs meeting consumers' expectations.

Due to environmental and ethical concerns, meat analogs represent an emerging trend to replace traditional animal meat. However, meat analogs lacking specific sensory properties (flavor, texture, color) would directly affect consumers' acceptance and purchasing behavior. In this review, we discussed the typical sensory characteristics of animal meat products from texture, flavor, color aspects, and sensory perception during oral processing. The related strategies were detailed to improve meat-like sensory properties for meat analogs. However, the upscaling productions of meat analogs still face many challenges (e.g.: sensory stability of plant-based meat, 3D scaffolds in cultured meat, etc.). Producing safe, low cost and sustainable meat analogs would be a hot topic in food science in the next decades. To realize these promising outcomes, reliable robust devices with automatic processing should also be considered. This review aims at providing the latest progress to improve the sensory properties of meat analogs and meet consumers' requirements.

Changes in odorants and flavor profile of heat-processed beef flavor during storage.

Flavor quality changes of heat-processed beef flavor (HPBF) upon storage are greatly affected by storage conditions. Flavor characterization of HPBF stored for 168 days resulted in 17 key odor-active compounds having flavor dilution factors log3 FD ≥ 3 and odor activity values  > 1. Based on the concentration changes of these key odorants, a total of 13 HPBF samples were divided into four groups by hierarchical cluster analysis. Combined with flavor profile analysis, the overall flavor of HPBF gradually changed from a characteristic beef aroma to burnt and fermented soybean paste-like notes. Some of the 17 key odorants, such as 2-methyl-3-furanthiol, 2-furfurylthiol, methyl furfuryl disulfide, and difurfuryl disulfide, contributed to the characteristic beef aroma of the freshly prepared product and in the early stage of storage. On the other hand, the odor-active compounds 2-methylbutanal, dimethyl disulfide, dimethyl trisulfide, and 2-ethyl-3,5-dimethylpyrazine were associated with the later stage of storage showing a positive correlation with the intensity of burnt and fermented soybean paste-like notes. A molecular understanding of flavor profile changes of HPBF during storage will provide the basis for developing characteristic beef aroma notes and maintaining the keeping quality of beef products upon storage. PRACTICAL APPLICATION: The relationship between sensory quality and changes in odorant concentration of heat-processed beef flavor during storage will provide the molecular basis for developing high-quality thermal reaction meat flavorings.

Investigating the Influence of Different Umami Tastants on Brain Perception via Scalp Electroencephalogram.

Three types of tastants are known as perceptually associated with umami taste: monosodium glutamate (MSG), disodium succinate (WSA), and disodium inosine monophosphate (IMP). While these tastants were confirmed to be perceptually similar in a sensory study, they could be discriminated (p < 0.05) by electroencephalogram (EEG) analysis on a time scale of 5-6 s. In comparison of the EEG responses of the participants, the brain could partly distinguish (p < 0.05) between different sensory intensities of MSG, WSA, or IMP. The EEG data indicated that the brain is partially sensitive to perceiving different sensory intensities (L, low; M, medium; and H, high) of the same umami stimuli; i.e., for MSG in µV2/Hz, L, 2.473 ± 0.181; M, 3.274 ± 0.181; and H, 3.202 ± 0.181. However, brain responses of perceptually equi-umami intensities could partially be discriminated, suggesting that the brain could partially discriminate (p < 0.05) MSG, WSA, and IMP, despite similar sensory intensities. Moreover, umami tastants were also found to significantly enhance (p < 0.05) the α wave activity, with the most responsive being at 10 Hz, particularly in the frontal and parietal and occipital regions of the brain (p < 0.001). This study shows the potential of EEG to investigate brain activity triggered by umami stimuli.

Application of gas chromatography-ion mobility spectrometry (GC-IMS) and ultrafast gas chromatography electronic-nose (uf-GC E-nose) to distinguish four Chinese freshwater fishes at both raw and cooked status.

The volatile organic compounds (VOCs) in four Chinese freshwater fishes (i.e., Hypophthalmichthys molitrix (H), Aristichthys nobilis (A), Lateolabrax japonicus (L), Parabramis pekinensis (P)) were separated using gas chromatography-ion mobility spectrometry (GC-IMS) and ultrafast gas chromatography electronic-nose (uf-GC E-nose). Principal component analysis (PCA) was applied to distinguish the VOCs identified from the four freshwater fishes in both raw and cooked states. Twenty compounds were identified from the spectral database of GC-IMS, including five aldehydes, eight alcohols, six ketones, and three esters. In addition, using GC E-nose, 32 compounds were isolated by the first column MTX-5, and 24 compounds were isolated by the second column MXT-1701. PCA results showed that the four fishes could be well discriminated against. The odor profiles of raw and cooked fishes were clearly different. This study demonstrated that specific signals provided from GC-IMS could differentiate freshwater fishes. GC-IMS and uf-GC E-nose could be developed further to distinguish aquatic products based on VOCs. PRACTICAL APPLICATIONS: Two new methods, gas chromatography-ion mobility spectrometry (GC-IMS) and ultrafast gas chromatography electronic-nose (uf-GC E-nose), were used to analyze the volatile organic compounds (VOCs) in four Chinese freshwater fishes at raw and cooked status. GC-IMS has the characteristics of fast detection speed and high sensitivity. The accuracy of the qualitative analysis of the compounds is better with GC-IMS (larger data volume, leading to a better in-depth statistical analysis). Uf-GC E-nose could provide a nondestructive, fast, relatively low cost, and trustworthy way for flavor analysis. According to the techniques, the established fingerprints of VOCs provided an additional tool for food analysis.

Saltiness-Enhancing Peptides Isolated from the Chinese Commercial Fermented Soybean Curds with Potential Applications in Salt Reduction.

Salt is very important for human health and food seasoning. Recently, several peptides isolated from natural food products have been reported exhibiting a salty taste or a saltiness-enhancing function. In this investigation, taste-active peptides occurring in commercial Chinese fermented soybean curd were isolated and identified using ultrafiltration, gel permeation chromatography, ion-exchange chromatography, and nano-LC/Q-TOF MS/MS. The salty taste-enhancing function of the target fractions was confirmed by both a rat taste cell model and/or human sensory evaluation. Four decapeptides were found as taste-active compounds. Among them, peptide E (EDEGEQPRPF) was the most potent saltiness-enhancing peptide: 0.4 mg/mL in 50 mmol/L NaCl solution could increase its salty perception equivalent to the salt level of 63 mmol/L NaCl reference solution. The sequence of the peptide has been found in the α'-subunit of ß-conglycinin [Glycine max]. The remaining peptides V (VGPDDDEKSW), DD (DEDEQPRPIP), and DG (DEGEQPRPFP) showed umami and kokumi tastes as well as a weak saltiness-enhancing sensation. These findings suggest that the decapeptide EDEGEQPRPF could be a possible alternative to partially reduce the amount of sodium intake without compromising for saltiness.

GC × GC-ToF-MS and GC-IMS based volatile profile characterization of the Chinese dry-cured hams from different regions.

Chinese dry-cured hams have unique aroma characteristics appreciated by local consumers. The volatile organic compounds (VOCs) of six selected Chinese dry-cured hams (Mianning, Nuodeng, Saba, Sanchuan, Wanhua, and Xuanen) were analyzed by solvent assisted flavor evaporation (SAFE) combined with GC × GC-ToF-MS and head-space (HS) injection combined with GC-IMS. To visualize VOCs and differentiate samples, principal component analysis (PCA) and multiple factor analysis (MFA) were performed. GC × GC-ToF-MS resulted in over five times more VOCs (265) than GC-IMS (45). However, PCA and MFA indicated similar results using GC-IMS or GC × GC-ToF-MS data, suggesting HS-GC-IMS as a good choice to differentiate dry-cured hams from different regions. Xuanen ham from Yunnan Province having smoky aroma was significantly different from other hams, likely due to its unique process. Many aldehydes (heptanal, nonanal, etc.) played an important role in Sanchuan ham. Ketones were related to other four dry-cured hams, though they came from different regions. This study provides valuable analytical data to characterize and discriminate the flavor profile of Chinese dry-cured hams.

Generation of α-Diketones and 4-Hydroxy-2,5-dimethyl-3(2H)-furanone upon Coffee Roasting-Impact of Roast Degree on Reaction Pathways.

The formation pathways of α-diketones (2,3-butanedione and 2,3-pentanedione) and 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) upon coffee roasting were investigated in a kinetic study applying labeled and unlabeled sucrose (CAMOLA approach) in biomimetic in-bean experiments. The results highlighted that not only did the contribution of sucrose to the level of α-diketones in roasted coffee change with the roasting degree but also the portion of the individual reaction pathways. At early roasting stages, 2,3-butanedione was formed from sucrose mainly via the intact sugar skeleton, whereas from the middle of the roasting course, the formation foremost occurred from sugar fragments, primarily by C1/C3 and C2/C2 recombinations. In contrast, 2,3-pentanedione was generated from sucrose mainly via an intact sugar skeleton during the whole roasting cycle; nevertheless, the share of 2,3-pentanedione formed by recombination of fragments (mainly C2/C3) progressively increased with roasting time. HDMF was generated from sucrose almost exclusively via cyclization of an intact skeleton, irrespective of the roast time.

Origin and yields of acetic acid in pentose-based Maillard reaction systems.

The formation of acetic acid from pentoses was studied in aqueous buffered systems (90-120 degrees C, pH 6.0-8.0) containing equimolar concentrations of 13C-labeled xylose and glycine. Acetic acid was quantified by gas chromatography-mass spectroscopy using an isotope dilution assay. Acetic acid was mainly formed from the C-1/C-2 carbon atoms of xylose (77-87%), while small amounts were also formed from the C-4/C-5 atoms of the pentose sugar (9-15%). Temperature and pH had only a small effect on the relative contribution of the sugar carbon atoms to acetic acid. These results support beta-dicarbonyl cleavage of 1-deoxypento-2,4-diulose as a major pathway leading to acetic acid in pentose-based Maillard reaction systems under food processing conditions. Acetic acid was confirmed as a major degradation product of pentoses at the early stage of the Maillard reaction, yielding 16 mol% and 28 mol% at pH 6.0 and pH 8.0, respectively.

Temporal changes in aroma release of Longjing tea infusion: interaction of volatile and nonvolatile tea components and formation of 2-butyl-2-octenal upon aging.

The temporal change in the headspace composition of an aroma model mimicking Longjing green tea aroma was studied in the presence of nonvolatile Longjing green tea constituents. Upon storage at 50 degrees C, the formation of 2-butyl-2-octenal was found, which increased with time. This enal was generated by crotonization of hexanal as demonstrated in model experiments. The formation of 2-butyl-2-octenal was also detected in Longjing tea infusions and Longjing tea leaves upon storage at 50 degrees C. The presence of nonvolatiles induced a strong decrease in aroma release. These effects were mainly due to catechins, major constituents of green tea infusion. Free amino acids, that is, glycine, contributed only to significantly decrease alpha,beta-unsaturated carbonyl aroma compounds, that is, 1-octen-3-one and geranial. Model reaction containing a mixture of 1-octen-3-one and glycine indicated on the basis of NMR and MS data the formation of the tentatively identified N-1-(3-oxo-octyl)glycine resulting from a 1,4-addition. The perceived aroma of green tea infusion is very likely to be affected by the formation of new aroma compounds and the changes in aroma release affected by interactions with tea nonvolatile components. This deserves further investigations on the sensory level.

Formation of furan and methylfuran by maillard-type reactions in model systems and food.

The formation of furan and 2-methylfuran was studied in model systems based on sugars and selected amino acids. Both compounds were preferably formed under roasting conditions in closed systems yielding up to 330 micromol of furan and 260 micromol of 2-methylfuran per mol of precursor. The amounts obtained under pressure cooking conditions were much lower, usually below 20 micromol/mol, except for 2-furaldehyde, which yielded 70-100 micromol/mol of furan. Labeling studies indicated two major formation pathways for both furans: (i) from the intact sugar skeleton and (ii) by recombination of reactive C(2) and/or C(3) fragments. Under roasting conditions in the absence of amino acids, furan was mainly formed from the intact sugar skeleton. Formic and acetic acid were identified as byproducts of sugar degradation, indicating the split off of C(1) and/or C(2) units from hexoses. The presence of alanine, threonine, or serine promoted furan formation by the recombination of C(2) fragments, such as acetaldehyde and glycolaldehyde, which may originate from both sugars and amino acids. In aqueous solution, about half of furan was generated by the recombination of sugar fragments. 2-Methylfuran was preferably formed in the presence of amino acids by aldol-type reactions of C(2) and C(3) fragments with lactaldehyde as a key intermediate, the Strecker aldehyde of threonine. The total furan levels in cooked vegetables were increased by spiking with hexoses. However, in pumpkin puree, only about 20% of furan was formed from sugars, preferably from the intact carbon skeleton.

New Insight into the Role of Sucrose in the Generation of α-Diketones upon Coffee Roasting.

The origin and formation pathways of the buttery-smelling α-diketones 2,3-butanedione and 2,3-pentanedione upon coffee roasting were studied by means of biomimetic in-bean experiments combined with labeling experiments. For this purpose natural sucrose in the coffee bean was replaced by fully or partially 13C-labeled sucrose or by a mixture of unlabeled and fully 13C-labeled sucrose (CAMOLA approach). The obtained data point out that sucrose contributes to both α-diketones; however, its importance and reaction pathways clearly differ. Whereas the major part of 2,3-pentanedione originates from sucrose (about 76%), its contribution to 2,3-butanedione is much lower (about 35%). Formation from intact sugar skeleton is the major pathway generating 2,3-pentanedione from sucrose, whereas 2,3-butanedione is mainly generated by recombination of sucrose fragments. The contribution of glucose and fructose moieties of sucrose to both α-diketones is comparable. Finally, kinetic experiments with fully labeled sucrose showed that the contribution of sucrose changes during roasting.

Model studies on the release of aroma compounds from structured and nonstructured oil systems using proton-transfer reaction mass spectrometry.

Relative retention, volatility, and temporal release of volatile compounds taken from aldehyde, ester, and alcohol chemical classes were studied at 70 degrees C in model systems using equilibrium static headspace analysis and real time dynamic headspace analysis. These systems were medium-chain triglycerides (MCT), sunflower oil, and two structured systems, i.e., water-in-oil emulsion and L2 phase (water-in-oil microemulsion). Hydrophilic domains of the emulsion type media retained specifically the hydrophilic compounds and alcohols. Four kinetic parameters characterizing the concentration- and time-dependent releases were extracted from the aroma release curves. Most of the kinetic parameter values were higher in structured systems than in oils particularly when using MCT. The oil nature was found to better control the dynamic release profiles than the system structures. The release parameters were well-related (i) to the volatile hydrophobicity as a function of the oil used and (ii) to the retention data in the specific case of the L2 phase due to a specific release behavior of alcohols.

Sustainable Food Processing Inspired by Nature.

Here, we elaborate on the natural origin and use of enzymes and cultures in sustainable food processing. We also illustrate how enzymatically treated or fermented food can contribute to solving challenges involving nutrition and health, such as aging, malnutrition, obesity, and allergy.