Motonaka Kuroda

Kokumi Substances, Enhancers of Basic Tastes, Induce Responses in Calcium-Sensing Receptor Expressing Taste Cells

Recently, we reported that calcium-sensing receptor (CaSR) is a receptor for kokumi substances, which enhance the intensities of salty, sweet and umami tastes. Furthermore, we found that several γ-glutamyl peptides, which are CaSR agonists, are kokumi substances. In this study, we elucidated the receptor cells for kokumi substances, and their physiological properties. For this purpose, we used Calcium Green-1 loaded mouse taste cells in lingual tissue slices and confocal microscopy. Kokumi substances, applied focally around taste pores, induced an increase in the intracellular Ca2+ concentration ([Ca2+]i) in a subset of taste cells. These responses were inhibited by pretreatment with the CaSR inhibitor, NPS2143. However, the kokumi substance-induced responses did not require extracellular Ca2+. CaSR-expressing taste cells are a different subset of cells from the T1R3-expressing umami or sweet taste receptor cells. These observations indicate that CaSR-expressing taste cells are the primary detectors of kokumi substances, and that they are an independent population from the influenced basic taste receptor cells, at least in the case of sweet and umami.

Effect of dried-bonito broth on mental fatigue and mental task performance in subjects with a high fatigue score

Dried-bonito broth is commonly employed as a soup and sauce base in Japanese cuisine and is considered to be a nutritional supplement that promotes recovery from fatigue. Previous human trials suggest that the ingestion of dried-bonito broth improves several mood states; however, its effect on fatigue has not yet been clarified. The aim of this study was to clarify the effects of daily ingestion of dried-bonito broth on fatigue and cognitive parameters by a placebo-controlled double blind crossover trial. Forty-eight subjects with fatigue symptoms ingested the dried-bonito broth or a placebo solution every day for 4 weeks. Mood states were evaluated by the Profile of Mood States (POMS), and mental task performance was evaluated by the Uchida-Kraepelin psychodiagnostic (UKP) test. Fatigue and total mood disturbance (TMD) scores on the POMS test decreased significantly during the dried-bonito broth ingestion (p<0.05), but did not change significantly during placebo ingestion. The change in vigor score during dried-bonito broth ingestion was significantly higher than that during placebo ingestion at 2 weeks (p<0.05). The results of the UKP test indicate that the numbers of both total answers and correct answers significantly increased during dried-bonito broth ingestion (p<0.05), while no significant changes were observed in the placebo ingestion. These results suggest that the daily ingestion of dried-bonito broth may improve the mood states, may reduce mental fatigue and may increase performance on a simple calculation task.

Determination and quantification of the kokumi peptide, γ-glutamyl-valyl-glycine, in commercial soy sauces

Recent studies have demonstrated that kokumi substances, such as glutathione, are perceived through the calcium-sensing receptor (CaSR), and screening by CaSR assay and sensory evaluation has shown that γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) is a potent kokumi peptide. In this study, the contents of γ-Glu-Val-Gly in six commercial brands of dark-coloured soy sauces, two brands of light-coloured soy sauce, and one brand of white soy sauce, were investigated by high performance liquid chromatography-tandem mass spectrometry (LC/MS/MS), followed by derivatization with 6-aminoquinoyl-N-hydroxysuccinimidyl-carbamate (AQC). The analyses indicated that γ-Glu-Val-Gly was present in all investigated soy sauces at concentrations ranging from 0.15 to 0.61mg/dl, demonstrating that it is widely distributed in soy sauces.

Determination and Quantification of γ-Glutamyl-valyl-glycine in Commercial Fish Sauces

It was recently reported that kokumi substances such as glutathione are perceived through the calcium-sensing receptor (CaSR). In addition, screening by the CaSR assay and sensory evaluation revealed that γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) was a potent kokumi peptide. In this study, the quantities of γ-Glu-Val-Gly in various commercial fish sauces originating from Vietnam (Nuoc Mum), Thailand (Nampra), China (Yu-lu), Korea, Japan (Shottsuru and Ikanago-shoyu), and Italy (Garum) were investigated using a LC/MS/MS method followed by derivatization with 6-aminoquinoyl-N-hydroxysuccinimidyl-carbamate (AQC). The analyses revealed γ-Glu-Val-Gly at concentrations ranging from 0.04 to 1.26 mg/dL, indicating that γ-Glu-Val-Gly is widely distributed among various commercial fish sauces.

Frequency of soup intake and amount of dietary fiber intake are inversely associated with plasma leptin concentrations in Japanese adults

Previous studies have shown that the intake of soup negatively correlates with the body mass index (BMI), suggesting that soup intake reduces the risk of obesity. In this study, to clarify the association of the intake of soup and various nutrients with plasma leptin concentration, a cross-sectional study on 504 Japanese adults aged 20-76 years (103 men and 401 women) was performed. The intake of soup and various nutrients was investigated by food frequency questionnaires. Plasma leptin concentration was measured in fasting blood by radioimmunoassay. The correlation was analyzed by multiple regression analysis. The average frequency of soup intake was 7.6 times/week. The average plasma leptin concentration was 7.76 ng/ml. After adjusting the confounding factors, the frequency of soup intake has a significant inverse association with plasma leptin concentration. Among the macronutrients, only dietary fiber intake negatively correlated with plasma leptin concentration after the adjustment for potential confounding factors. These results suggest that the intakes of soup and dietary fiber were negatively correlated with plasma leptin concentration in Japanese adults.

Determination of γ-glutamyl-valyl-glycine in raw scallop and processed scallop products using high pressure liquid chromatography-tandem mass spectrometry.

The determination of the kokumi peptide, γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) in raw scallop and processed scallop products was carried out using high pressure liquid chromatography-tandem mass spectrometry (LC/MS/MS). The detection of γ-Glu-Val-Gly was achieved using a multiple reaction monitoring (MRM) method. The optimised condition enabled the precise determination of γ-Glu-Val-Gly. Raw scallop contained 0.08 μg/g γ-Glu-Val-Gly, and the γ-Glu-Val-Gly levels in processed scallop products, such as dried-scallop and scallop extract, were measured to be 0.64 and 0.77 μg/g, respectively. This is the first report to confirm the existence of γ-Glu-Val-Gly in foodstuff.

Mechanism of the perception of “kokumi” substances and the sensory characteristics of the “kokumi” peptide, γ-Glu-Val-Gly

Some foods are known to have flavours that cannot be explained by the five basic tastes alone, such as continuity, mouthfulness and thick flavour. It was demonstrated that these sensations are evoked by the addition of kokumi substances, flavour modifiers that have no taste themselves. However, their mode of action has been poorly understood. During a study on the perception of amino acids and peptides, it was found that glutathione (GSH) was one of the agonists of the calcium-sensing receptor (CaSR). We have hypothesized that CaSR is involved in the perception of kokumi substances. We found that all CaSR agonists tested act as kokumi substances and that a positive correlation exists between the CaSR activity of γ-glutamyl peptides and kokumi intensity. Furthermore, the kokumi intensities of GSH and γ-Glu-Val-Gly, a potent kokumi peptide, were significantly reduced by the CaSR-specific antagonist, NPS-2143. These results suggest that CaSR is involved in the perception of kokumi substances. A potent kokumi peptide, γ-Glu-Val-Gly, enhanced sweetness, saltiness and umami when added to 3.3% sucrose, 0.9% NaCl and 0.5% MSG solutions, respectively. In addition, γ-Glu-Val-Gly enhanced the intensity of continuity, mouthfulness and thick flavour when added to chicken soup and reduced-fat cream. These results suggest that γ-Glu-Val-Gly is a potent kokumi peptide and would be useful for improving the flavour of food.

Frequency of Soup Intake Is Inversely Associated with Body Mass Index, Waist Circumference, and Waist-to-Hip Ratio, but Not with Other Metabolic Risk Factors in Japanese Men

Several previous studies have shown that the intake of soup negatively correlates with the body mass index (BMI), serum cholesterol and triacylglycerol levels, and blood pressure, suggesting that soup intake reduces metabolic risk. However, the correlation between soup intake and various metabolic risk factors has not been well-established. Especially, it has not been investigated in Asian countries. The aim of this study was to investigate the association of the frequency of soup intake and metabolic risk factors such as BMI, waist circumference, waist-to-hip ratio, serum cholesterol, serum triacylglycerol, blood glucose, and glycated hemoglobin. A cross-sectional study of 103 Japanese men aged 24 to 75 years was conducted. The intake of soup and other food was investigated by semi-quantitative food frequency questionnaires. The correlation between the frequency of soup intake and metabolic risk factors was analyzed by multiple regression analysis with a linear model. The median value of frequency of soup intake was 7.0 times per week. After adjusting for confounding factors such as age, energy intake, energy from alcohol intake, current smoking, and estimated energy expenditure, the frequency of soup intake was found to have a significant inverse association with BMI (P=0.040), waist circumference (P=0.024), and waist-to-hip ratio (P=0.001). However, no significant associations with other metabolic risk factors were found. Frequency of soup intake is negatively correlated with obesity-related physical parameters in Japanese men.

Effect of a kokumi peptide, γ-glutamyl-valyl-glycine, on the sensory characteristics of chicken consommé

BackgroundRecent studies have demonstrated that kokumi substances such as glutathione are perceived through the calcium-sensing receptor (CaSR). Screening by a CaSR assay and sensory evaluation have shown that γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) is a potent kokumi peptide. In the present study, the sensory characteristics of chicken consommé with added γ-Glu-Val-Gly were investigated using descriptive analysis.ResultsChicken consommé containing γ-Glu-Val-Gly had significantly stronger “umami” and “mouthfulness” (mouth-filling sensation) characteristics than the control sample at a 99% confidence level and significantly stronger “mouth-coating” characteristic than controls at a 95% confidence level.ConclusionsThese data suggest that a kokumi peptide, γ-Glu-Val-Gly, can enhance umami, mouthfulness, and mouth coating, implying that the application of this peptide could contribute to improving the flavor of chicken consommé.

The Effect of the Dried-Bonito Broth on Blood Pressure, 8-Hydroxydeoxyguanosine (8-OHdG), an Oxidative Stress Marker, and Emotional States in Elderly Subjects.

Dried-bonito broth (DBB, katsuo-bushi dashi) is commonly used in Japanese cuisine, and is also used as a traditional remedy for recovery from fatigue and improvement of blood circulation. To clarify the effect of DBB on blood pressure, oxidative stress and emotional states, a randomized crossover human trial was performed. Twenty-seven elderly Japanese subjects ingested DBB or water for one month. Measurement of blood pressure and urinary 8-hydroxydeoxyguanosine (8-OHdG) and evaluation of emotional states were performed before and after the ingestion periods. The changes in systolic blood pressure (SBP) during DBB ingestion was significantly lower than that during water ingestion (p = 0.037). Urinary 8-OHdG significantly decreased during DBB ingestion (p = 0.0002). Evaluation of emotional states indicated that composure significantly improved during DBB ingestion (p = 0.034). These results suggest that the daily ingestion of DBB lower SBP, reduce urinary 8-OHdG and might improve emotional states in elderly subjects.