Multienzyme-Mediated Dual-Channel Magnetic Relaxation Switching Taste Biosensor (D-MRSTB) for Simultaneous Detection of Umami Compounds and Synergistic Enhancement in Food.

Umami substances play a significant role in the evaluation of food quality, and their synergistic enhancement is of great importance in improving and intensifying food flavors and tastes. Current biosensors available for umami detection still confront challenges in simultaneous quantification of multiple umami substances and umami intensities. In this study, an innovative dual-channel magnetic relaxation switching taste biosensor (D-MRSTB) was developed for the quantitative detection of representative umami substances. The multienzyme signal of D-MRSTB specifically catalyzes the umami substances of interest to generate hydrogen peroxide (H2O2), which is then used to oxidate Fe2+ to Fe3+. Such a valence-state transition of paramagnetic ions was utilized as a magnetic relaxation signaling switch to influence the transverse magnetic relaxation time (T2) within the reaction milieu, thus achieving simultaneous detection of monosodium glutamate (MSG) and inosine 5'-monophosphate (IMP). The biosensor showed good linearity (R2 > 0.99) in the concentration range of 50-1000 and 10-1000 µmol/L, with limits of detection (LOD) of 0.61 and 0.09 µmol/L for MSG and IMP, respectively. Furthermore, the biosensor accurately characterized the synergistic effect of the mixed solution of IMP and MSG, where ΔT2 showed a good linear relationship with the equivalent umami concentration (EUC) of the mixed solution (R2 = 0.998). Moreover, the D-MRSTB successfully achieved the quantitative detection of umami compounds in real samples. This sensing technology provides a powerful tool for achieving the detection of synergistic enhancement among umami compounds and demonstrates its potential for application in the food industry.

Physicochemical properties as indicators of sensory characteristics of jidori and broiler chicken meat.

To improve the eating quality of chicken, the physicochemical properties were examined, which serve as indicators for breeding. Thigh meat was collected from 384 chickens from seven and nine types of the jidori (free-range local traditional pedigree chickens) or broiler chickens, respectively. The principal component analysis of the physicochemical values of the jidori and broilers were arranged as different groups in the score plot. The results of multiple regression analysis of the sensory characteristics and physicochemical properties of thigh meat indicated that the tenderness decreased with the higher crude protein content and cooking loss in the jidori and increased with lower cooking loss and higher moisture and ether extract content in the broiler. The juiciness of the jidori decreased as the moisture content decreased, and that of the broiler decreased as the cooking loss and crude protein content increased. The umami of both the jidori and broiler was improved by increasing the 5'-inosinic acid content. Therefore, it was suggested that the physicochemical properties, which serve as indicators of the eating quality of chicken, differed between the jidori and broiler, and that the general biochemical components, cooking loss and 5'-inosinic acid content may serve as breeding indicators.

Inosine 5'- monophosphate derived umami taste intensity of beef determination by electrochemistry and chromatography.

The umami sensation contributes to beef taste and acceptability. Inosine 5'- monophosphate (IMP), the most abundant 5'-ribonucleotide in meat, is known to impart an umami taste without the undesired side effects commonly associated with glutamate. Nevertheless, the investigation of IMP's role in beef flavor has thus far been overlooked. Traditional methods for detecting IMP have relied on liquid chromatography coupled with ultraviolet spectroscopy or mass spectrometry techniques. However, these methods are not practical for production settings due to the complexity and resource demands of sophisticated laboratory techniques. Alternative methods like cyclic voltammetry might offer more practical solutions for rapidly detecting IMP. The objectives of this study were to evaluate the efficiency of using electrochemistry and chromatography on differentiating beef strips spiked with different IMP contents. The IMP threshold was 0.30 mM determined by a trained panel using the Best Estimates Threshold method. Beef strip steaks of USDA Prime, Choice, and Select were spiked at 0.30 and 0.60 mM of IMP, based on green weight and an estimated moisture content of 65%. In this study, differences in the IMP content of steaks were not detected by liquid chromatography-mass spectrometry. However, the cyclic voltammetry approach differentiated IMP concentrations at 0.50 mM or above in aqueous solutions and subsequentially meat extracts from the buffered blank solutions. In conclusion, cyclic voltammetry holds potential as a rapid and effective approach for detecting IMP in beef and other meat products, offering promising applications for future research.

A comparative study of meat quality and vibrational spectroscopic properties of different chicken breeds.

Chicken breed is one of the key factors that influence meat quality. The quality attributes of breast meat from commercial broiler (CB), Thai native chicken (NC, Leung Hang Khao), and the crossbred Korat chicken (KC) were investigated via synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy, Fourier transform Raman (FT-Raman) spectroscopy, and physicochemical analysis. The protein and carbonyl contents of KC and NC meats were higher than that of CB meat, but the lipid content was lower (P < 0.05). CB meat was characterized by high moisture, lightness (L*), and presence of taste-active nucleotides, namely, inosine 5'-monophosphate (IMP) and guanosine 5'-monophosphate (GMP). Moreover, NC meat had the highest insoluble collagen and inosine contents (P < 0.05). The predominant protein secondary structures of KC and NC meats were ß-turns and random coils, whereas α-helices were mainly found in CB meat. Based on principal component analysis, the meat quality and spectra were clearly separated by breeds. The high moisture and lipid content of meat corresponded to O-H stretching (3,203 cm-1) and C-H stretching (2,854 cm-1) in the FT-Raman spectra, whereas PO2- stretching (1,240 cm-1), measured via SR-FTIR, was well correlated with the IMP content. In addition, the FT-Raman wavenumber of 934 cm-1, indicating C-C stretching, was correlated with high water-holding capacity (WHC) in KC meat. The quality of meat from slow- and fast-growing chickens significantly varies. Vibrational spectroscopy is a powerful technique that provides insightful molecular information correlated with various meat attributes.

Screening out important substances for distinguishing Chinese indigenous pork and hybrid pork and identifying different pork muscles by analyzing the fatty acid and nucleotide contents.

Fatty acids and nucleotides are major contributors of pork flavor, which are key quality traits influencing consumers' preference. In this study, we investigated the contents of nucleotides (inosinic acid, inosine, and hypoxanthine) and 26 fatty acids by high-performance liquid chromatography and gas chromatography in longissimus dorsi and semimembranous muscle of four breeds (Dongbei Min pig, Beijing Black pig, Wuzhishan pig, and DLY pigs). Combining with multivariate statistics, we found significant differences in the nucleotides and fatty acids compositions among the breeds and muscles. Furthermore, the content of total unsaturated fatty acids and DHA were higher in native pigs than crossbred pigs. This study provides a theoretical basis for understanding the composition of flavor precursor substances, as well as for the identification of different muscles.

Meat quality and Raman spectroscopic characterization of Korat hybrid chicken obtained from various rearing periods.

Meat quality attributes vary with chicken age. Understanding the relationship between poultry age and the quality of the meat would be beneficial for efficient poultry farming to meet market needs. The Korat hybrid chicken (KC) is a new crossbred chicken whose meat quality is distinct from that of commercial broiler (CB) chickens and has not been well characterized. In this study, we characterized the physico-chemical properties of KC meat and correlate the findings with Raman spectral data. The protein content of KC breast and thigh meat increased with age. The pH of thigh meat decreased, while the water-holding capacity of breast meat increased as the age of the chickens increased. The amount of cholesterol in breast meat decreased as the rearing period was extended. Inosine 5'-monophosphate and guanosine 5'-monophosphate of breast meat decreased as KC grew older. The shear force values of meat from older birds increased concomitantly with an increase in total collagen. Principle component analysis revealed that the meat quality of CB was greatly different from that of KC meat. High shear force values of KC meat at 20 wk of age were well correlated with an increase in the ß-sheet structure (amide I) and amide III of collagen. Raman spectra at 3,207 cm-1 and relative α-helical content were negatively correlated with shear force values of KC breast meat. These could be used as markers to evaluate KC meat quality.

Brackish water improves the taste quality in meat of adult male Eriocheir sinensis during the postharvest temporary rearing.

We investigated the effect of temporary rearing in brackish water on the taste quality in meat of crab cooked. The main salinity-responsive factors included 5'-nucleotides and free amino acids (FAAs) in crab meat that were identified using tri-step infrared spectroscopy. Compared to the fresh water group, the contents of 5'-adenosine monophosphate and 5'-inosine monophosphate in the brackish water group significantly increased in the 2nd week and decreased in the 6th week, respectively. The contribution ratio of umami FAAs increased from 8.1 to 13.5% in the 4th week in the brackish water group, showing maximum value of equivalent umami concentration. Moreover, Ca2+ and Cl- contents significantly increased in the 4th and 6th weeks, respectively (P < 0.05). Infrared spectroscopy was an effective method to identify the taste components. With respect to the taste quality, four weeks were determined as the best period for temporary rearing of the crab in brackish water.

Temporal profile of flavor enhancers MAG, MSG, GMP, and IMP, and their ability to enhance salty taste, in different reductions of sodium chloride.

We evaluated the temporal profile of the flavor enhancers monosodium glutamate (MSG), disodium inosinate (IMP), disodium guanylate (GMP), and monoammonium glutamate (MAG). We also evaluated the ability of these flavor enhancers to enhance salty taste in solutions containing different reductions of sodium chloride. Four experiments were conducted using Central Composite Rotational Design (CCRD) with focus on two objectives: concentration of flavor enhancers (0% to 1%) and reduction of sodium chloride content (0% to 100%). A 0.75% saline solution of NaCl was used as a control. In each experiment, the treatments were evaluated by the intensity of salty and umami tastes using an intensity scale. Treatments, selected according to the results of CCRD, were analyzed using time-intensity (TI) and temporal dominance of sensations (TDS) analyses. Glutamates (MSG/MAG) showed greater capacity to enhance salty taste than treatments containing nucleotides (IMP/GMP). The intensity of umami taste, using all the examined flavor enhancers, showed a similar sensory profile. Temporal perception curves (TI and TDS) of salty and umami tastes also showed a similar temporal profile. The glutamic acid amino acids were better able to improve salty taste than nucleotides in any range of sodium chloride reduction. Flavor enhancers showed greater ability to increase salty taste in smaller reductions in sodium chloride content. PRACTICAL APPLICATION: This research expand the knowledge about the ability to enhance the salty taste of flavor enhancers in different reductions in sodium content, Beside that, will provide information about the time profile of flavor enhancers. This study provides scientific technical information on the ability to intensify the salty taste of flavor enhancers and can assist the industry to develop new low sodium products and encourage the scientific community to conduct future research on this subject.

Use of Hermetia illucens larvae as a dietary protein source: Effects on growth performance, carcass traits, and meat quality in finishing pigs.

This study investigated the effects of feeding Hermetia illucens larvae (0, 4, and 8%; HI0, HI4, and HI8 groups, respectively) on growth performance, carcass traits, and meat quality of finishing pigs. Results showed that the HI4 diet increased (P < .05) final body weight and average daily gain and decreased (P < .05) feed to gain ratio compared with HI0 and HI8 group. HI4 and HI8 diets increased (P < .05) loin-eye area, marbling scores, and inosine monophosphate content of longissimus thoracis (LT) compared with HI0 diet. The intramuscular fat content was greater (P < .05) in HI4 group than in the HI0 group. Furthermore, HI4 diet up-regulated (P < .05) lipogenic genes and MyHC-IIa mRNA levels in LT compared with HI0 diet. Our results indicated that dietary inclusion of H. illucens larvae has a beneficial impact on growth performance and meat quality, and the underlying mechanism may be due to the altered lipogenic potential induced by H. illucens larvae.

Evaluation of the four breeds in synthetic line of Jiaxing Black Pigs and Berkshire for meat quality traits, carcass characteristics, and flavor substances.

To investigate breed characteristics and the effect of hybridization of Jiaxing Black Pig (JBP) with Western breeds, the carcass and meat quality traits and flavor substances such as inosinic acids (IMP), intramuscular fat (IMF) in longissimus muscle (LM) from five breeds including JBP, Berkshire, Berkshire × JBP (BJBP), Duroc × Berkshire × JBP (DBJBP), Duroc × Landrace × JBP (DLJBP) were compared in this study. It was found that water holding capacity (WHC) of LM in JBP was significantly higher than that in the other strains (p < 0.01). Dressing out percentage and lean percentage of JBP were both significantly lower than those in the others (p < 0.01) in connection with their lighter carcass weight and higher subcutaneous fat percentage (p < 0.01). Heterosis was realized in DJBP, DBJBP, and DLJBP since their carcass weight, lean percentage, and loin eye muscle area (LEMA) were markedly higher when compared to JBP, whereas lower than those in Berkshire. Among the breeds, the content of IMF and IMP in the LM of JBP were the highest. These traits were also palpably improved in the crossbreds, especially for DBJBP, of which pork was considered outstanding for containing the most abundant essential amino acids (EAA) and total amino acids (TAA).

Superchilled storage (-2.5 ± 1°C) extends the retention of taste-active and volatile compounds of yellow-feather chicken soup.

This work investigated the effects of refrigerated storage (RS: 4 ± 1°C) and superchilled storage (SS: -2.5 ± 1°C) on non-volatile and volatile compounds in chicken soup made from Chinese yellow-feather broilers. The results from total viable count (TVC) and coliform analysis showed that soups were safe for human consumption after a storage period of 42 days. SS resulted in a significantly (p < .05) higher content of free amino acids (umami and sweet taste) and 5'-nucleotides (inosine 5'-monophosphate and adenosine 5'-monophosphate) from 21 to 42 days compared to RS. Hexanal, (E)-2-decenal, (E,E)-2,4-decadienal and 2-pentyl furan were described as the primary odorants. SS showed significantly lower values (p < .05) for ketones and hydrocarbons, higher values for aldehydes and alcohols from 14 to 42 days, when compared to RS. The results suggest that SS improved the flavor retention of chicken soup after 21 days of storage and is a potential alternative treatment compared to RS.

Development and validation of a HILIC-UV method for the determination of nucleotides in fish samples.

The aim of this work was the development of a simple, novel and accurate method for the determination of adenosine triphosphate (ATP) and its first five catabolites: adenosine diphosphate (ADP), adenosine monophosphate (AMP), inosine monophosphate (IMP), inosine (Ino) and hypoxanthine (Hx), in fish tissue, based on hydrophilic interaction liquid chromatography (HILIC). For this purpose, a stationary phase for polar and hydrophilic compounds (ZIC-pHILIC) was used. The effect of different chromatographic parameters and the molecular mechanism based on the van't Hoff plot were examined. The t-test and Dixon's Q-test were applied in order to examine statistical differences and outlier values. The recovery of the method ranged between 82.7% and 127% and the %RSD values were lower than 10% for all analytes determined. The method was applied in frozen sea bream samples stored at 0-4 °C. The Ki-, G-, H- and F values were calculated for the estimation of the level of fish freshness.

Characteristic Flavor of Traditional Soup Made by Stewing Chinese Yellow-Feather Chickens.

The traditional recipe for Chinese chicken soup creates a popular taste of particular umami and aroma. The present study investigated the effects of stewing time (1, 2, and 3 h) on the principal taste-active and volatile compounds and the overall flavor profile of traditional Chinese chicken soup by measuring the contents of free amino acids (FAAs), 5'-nucleotides, minerals and volatile compounds and by evaluating the taste and aroma profiles using an electronic nose, an electronic tongue and a human panel. Results showed that the major umami-related compounds in the chicken soup were inosine 5'-monophosphate (IMP) and chloride, both of which increased significantly (P < 0.05) during stewing. The taste active values (TAVs) of the equivalent umami concentration (EUC) increased from 4.08 to 9.93 (P < 0.05) after stewing for 3 h. Although the FAA and mineral contents increased significantly (P < 0.05), their TAVs were less than 1. The volatile compounds were mainly hexanal, heptanal, octanal, nonanal, (E)-2-nonanal, (E)-2-decenal, (E,E)-2,4-decadienal, 1-hexanol, and 2-pentyl furan. With the prolonged stewing time, the aldehydes first increased and then decreased significantly (P < 0.05), while 1-hexanol and 2-pentyl furan increased steadily (P < 0.05). The aroma scores of the chicken soup reached the maximum after stewing for 3 h. The discrepancy in overall flavor characteristics tended to stabilize after 2 h of stewing. In general, stewing time has a positive effect on improving the flavor profiles of chicken soup, especially within the first 2 h.

Search for an index for the taste of Japanese Black cattle beef by panel testing and chemical composition analysis.

To search for an index for chemical composition related to superior taste in Japanese Black beef, we conducted panel tests and analyzed the chemical composition of seven beef brands. Thirty-five sirloin beefs from five heifers were used in this study, sold under seven beef brands graded as more than A4 on the Japanese Meat Grade scale. The chemical composition analyses assessed both raw and roasted meat, the latter of which was roasted under the same conditions as those used for the panel test. Results of the panel test and chemical composition analyses revealed that fatty acid composition, sugar content, adenosine triphosphage (ATP)-related compounds, amino acid composition and odor composition in the sirloin meat differed among beef brands. Furthermore, the correlations of chemical compositions between roasted and raw meat were significantly high. Sugar content and ATP-related compounds in roasted meat were significantly correlated with the item 'overall evaluation' of the panel test. ATP-related compounds, such as inosinic acid, carnosine and taurine, in roasted and raw meat were correlated significantly with the item 'umami intensity' of the panel test. These results suggest that the composition of these components is important for an index related to the overall evaluation of beef.

Enzymatic production of 5'-inosinic acid by AMP deaminase from a newly isolated Aspergillus oryzae.

5'-adenylic acid deaminase (AMP deaminase), an important enzyme for the food industry, can catalyze the irreversible hydrolysis of adenosine monophosphate (AMP) to inosine monophosphate (IMP) and ammonia. In this study, a new strain was screened that efficiently produces 3191.6U/g of AMP deaminase at 32°C. After purification, the optimal temperature and pH of the AMP deaminase were found to be 40°C and 6.0, respectively, but it was partially inhibited by Fe(3+), Cu(2+), Al(3+), and Zn(2+). With amplification of the AMP deaminase production system, 6mL of crude enzyme could produce 2.00mg/g of IMP from 2.04mg/g of dried yeast with an 84.8% molar yield after 40min. These results provide a new insight into AMP deaminase production and offer a potential platform for producing 5'-IMP.

Insertion of a knockout-first cassette in Ampd1 gene leads to neonatal death by disruption of neighboring genes expression.

AMPD1 is an adenosine monophosphate deaminase that catalyzes the deamination of AMP to IMP. To understand the physiological function of AMPD1, we obtained a strain of Ampd1 mutant mice from KOMP repository, which was generated by a knockout-first strategy. An elevated AMP level and almost complete lack of IMP was detected in the skeletal muscle of E18.5 Ampd1tm1a/tm1a mice. However, Ampd1tm1a/tm1a mice died in 2 days postnatally, which was contradicting to previous reports. After removal of the knockout-first cassette and critical exon, mice homozygous for the Ampd1tm1c/tm1c and Ampd1tm1d/tm1d alleles survived to adulthood. RNA-seq analysis indicated that the expression of two neighboring genes, Man1a2 and Nras, were disrupted in the Ampd1tm1a/tm1a mice, but normal in the Ampd1tm1c/tm1c and Ampd1tm1d/tm1d mice. The neonatal lethality phenotype in the Ampd1tm1a/tm1a mice was consistent with the Man1a2-deficient mice. Our results indicated the knockout-first cassette may cause off-target effect by influence the expression of neighboring genes. This study, together with other reports, strongly suggests that removal of targeting cassette by site-specific recombinases is very important for the accurate phenotypic interpretation on mice generated by target mutations.

Effects of different concentrations of metal ions on degradation of adenosine triphosphate in common carp (Cyprinus carpio) fillets stored at 4°C: An in vivo study.

The impact of different concentrations of Na(+), K(+), Ca(2+), Mg(2+), Fe(2+), and Zn(2+) on the degradation of adenosine triphosphate (ATP) and the influence of these ions on the activity of adenosine monophosphate deaminase (AMP-deaminase) and acid phosphatase (ACP) in common carp fillets (in vivo) during 4°C storage was examined. The content of ATP, inosine monophosphate (IMP), and hypoxanthine (Hx), and the activity of AMP-deaminase and ACP were determined. Results indicated that the effects of different concentrations of six kinds of metal ions on AMP-deaminase and ACP were not the same. Na(+), K(+), Fe(2+), and Zn(2+) enhanced AMP-deaminase activity, which led to the rapid degradation of ATP and to the generation of a large quantity of IMP within a short time. Ca(2+) and Mg(2+) delayed the change in AMP-deaminase and ACP activity in carp and caused a further delay in the degradation of ATP. Fe(2+) and Zn(2+) inhibited ACP activity, which reduced the decomposition of IMP and the formation of Hx.

Effect of Sugar on the Changes in Quality of Lightly Salted Grass Carp (Ctenopharyngodon idellus) Fillets under Vacuum Packaging at 4°C.

To estimate the effect of a low concentration of sugar on the changes in quality of lightly salted grass carp (Ctenopharyngodon idellus) during storage under vacuum packaging at 4°C, we determined the sensory score, total viable counts, biochemical quality, and physical quality of fish fillets. Fish samples were left untreated, dry cured with 1.3% salt, or dry cured with 1.3% salt plus 1.0% sugar. Compared with untreated samples, curing treatments reduced chemical changes reflected in pH, inosine monophosphate, hypoxanthine riboside, hypoxanthine, and total volatile base nitrogen; decreased the formation of phenylethylamine, putrescine, cadaverine, and histamine; and increased the overall sensory quality of fillets (P < 0.05). Compared to dry cured with 1.3% salt samples, sugar treatment significantly inhibited (P < 0.05) the increase in pH and total volatile base nitrogen value, but it promoted microbial growth and the formation of phenylethylamine and tyramine at later stages of storage. By considering each indicator, the addition of sugar, which can improve the taste of fillets, has no significant effect on the shelf life of vacuum-packaged grass carp fillets.

Changes in taste compounds, breaking properties, and sensory attributes during dry aging of beef from Japanese black cattle.

Analysis of the quality of highly marbled beef during dry aging for 60days after slaughter showed that the changes in some qualities differed from those of conventional meat. The tenderness of these meats did not change during aging for 50days but then gradually increased until day 60. The juiciness of these meats, as determined by sensory evaluation, did not change during aging for 60days, except for a decrease on day 20. The umami intensity of these meats in the sensory evaluation and the value calculated by Glu and IMP quantification were highest on day 40. This high umami intensity was induced by the synergistic effect of umami compounds such as Glu and IMP. These results for tenderness, juiciness, umami intensity, and flavor intensity suggested that the best duration of dry aging for highly marbled beef was 40days.

Umami compounds enhance the intensity of retronasal sensation of aromas from model chicken soups.

We examined the influence of taste compounds on retronasal aroma sensation using a model chicken soup. The aroma intensity of a reconstituted flavour solution from which glutamic acid (Glu), inosine 5'-monophosphate (IMP), or phosphate was omitted was significantly lower (p<0.05) than that of the model soup. The aroma intensity of 0.4% NaCl solution containing the aroma chicken model (ACM) with added Glu and IMP was significantly higher (p<0.05) than that of 0.4% NaCl solution containing only ACM. The quantitative analyses showed that adding monosodium glutamate (MSG) to aqueous aroma solution containing only ACM enhanced the intensity of retronasal aroma sensation by 2.5-folds with increasing MSG concentration from 0% to 0.3%. Sensation intensity using an umami solution with added MSG and IMP was significantly higher than that with only MSG when the MSG concentration was 0.05%, 0.075%, or 0.1%. However, it plateaued when MSG concentration was beyond 0.3%.