Diversity of γ- glutamyl peptides and oligosaccharides, the "kokumi" taste enhancers, in seeds from soybean mini core collections.

Soybeans (Glycine max (L,) Merr,) contain γ-glutamyl peptides and oligosaccharides, and these components play an important role in imparting the "kokumi" taste to foods. To gain insight into the genetic diversities and molecular mechanisms of accumulation of γ-glutamyl peptides and oligosaccharides in soybean, we measured the contents of these components using the Japan and World mini core collections. Similar to other previously reported traits, wide variations were detected among the accessions in the core collections with respect to the content of γ-glutamyl peptides and oligosaccharides. We found a positive relationship between the content of γ-glutamyl tyrosine and γ-glutamyl phenylalanine and between the content of raffinose and stachyose. Furthermore, there were unique accessions that included high levels of γ-glutamyl peptides and oligosaccharides. These accessions may be helpful in understanding the accumulation mechanism of γ-glutamyl peptides and oligosaccharides and to increase the "kokumi" taste components in soybean by performing a genetic analysis.

Isolation and characterization of key contributors to the "kokumi" taste in soybean seeds.

The water extract of soybean seeds (Glycine max (L.) Merr.) is nearly tasteless, but "kokumi" taste sensation was confirmed upon addition of a basic umami solution containing glutamic acid, inosine monophosphate, and sodium chloride. To identify the key contributors to the "kokumi" taste sensation in soybean seeds, sensory-guided fractionation, taste sensory analyses, and LC-MS/MS analyses were utilized. γ-glutamyl-tyrosine and γ-glutamyl-phenylalanine were identified as contributors to "kokumi taste"; specifically, these γ-glutamyl peptides imparted the "kokumi" taste sensation at a low taste threshold in a basic umami solution. Raffinose and stachyose, which are sufficiently present in soybean seeds, exhibited a synergistic effect in regard to the enhanced "kokumi" taste sensation of γ-glutamyl peptides. This is the first report that the combined use of γ-glutamyl peptides and oligosaccharides can increase the "kokumi" intensity, which suggests that soybean extracts or soymilk can be used to enhance the "kokumi" taste sensation in food products.

Absorption of the indigestible disaccharide, ß-1,4-mannobiose, from coconut by the rat portal vein.

The intestinal absorption of ß-1,4-mannobiose by rats was investigated. Mannobiose was detected in the portal vein plasma by matrix-assisted laser desorption ionization/time of flight mass spectrometry after its administration to rats. The presence of mannobiose in the rat plasma was confirmed by an experiment using ß-mannosidase. These results indicate that mannobiose was directly absorbed through the intestines even without being hydrolyzed.

Beta-conglycinin lowers very-low-density lipoprotein-triglyceride levels by increasing adiponectin and insulin sensitivity in rats.

The relationship between insulin sensitivity and the plasma triglyceride-lowering effect induced by beta-conglycinin was investigated. Male Wistar rats (19 weeks old) were fed diets containing casein, soy protein isolate, or beta-conglycinin for 4 weeks. In oral glucose administration, the beta-conglycinin-fed rats showed a significant decrease in the area under the glucose curve (0-60 min) as compared with the casein-fed rats. The hypoglycemic effect was significantly higher in the beta-conglycinin-fed rats than in the casein-fed rats at 30 min after intraperitoneal insulin injection. The liver sterol regulatory element-binding-protein-1 mRNA expression level was significantly lower and the plasma adiponectin concentration was significantly higher in the beta-conglycinin-fed rats than in the casein-fed rats. The hypotriglyceridemic effect of beta-conglycinin depended on a significant decrease in the concentration of very-low-density-lipoprotein triglycerides. These results indicate that beta-conglycinin increases adiponectin levels and improves glucose tolerance. The ability of beta-conglycinin to lower plasma lipid levels might be due to increased insulin sensitivity of the liver.

Structured triacylglycerol containing behenic and oleic acids suppresses triacylglycerol absorption and prevents obesity in rats.

BACKGROUND: Dietary 1(3)-behenoyl-2,3(1)-dioleoyl-rac-glycerol (BOO) has been reported to inhibit pancreatic lipase activity in vitro and suppress postprandial hypertriacylglycerolemia in humans. In the present study, the anti-obesity activities of BOO and its inhibitory effects on lymphatic triacylglycerol (TAG) absorption were investigated in rats. METHODS: In Experiment 1, rats were fed either BOO or soybean oil (SO) diet for 6 weeks. In the BOO diet, 20% of SO was replaced with an experimental oil rich in BOO. In Experiments 2 and 3, rats cannulated in the thoracic duct were administered an emulsions containing trioleoylglycerol (OOO) or an oil mixture (OOO:BOO, 9:1). Tri[1-14C]oleoylglycerol (14C-OOO) was added to the emulsions administered in Experiment 3. RESULTS: No observable differences were detected in food intake or body weight gain between the BOO and SO groups in Experiment 1. Plasma and liver TAG concentrations and visceral fat weights were significantly lower in the BOO group than in the SO group. The apparent absorption rate of fat was significantly lower in the BOO group than in the SO group. In Experiment 2, the lymphatic recovery of oleic and behenic acids was significantly lower at 5 and 6 h after BOO administration than after OOO administration. In Experiment 3, the lymphatic recovery of 14C-OOO was significantly lower at 5 and 6 h after BOO administration than after OOO administration. CONCLUSIONS: These results suggest that BOO prevents deposition of visceral fat and hepatic TAG by lowering and delaying intestinal absorption of TAG.

Effects of heat treatment under low moisture conditions on the protein and oil in soybean seeds.

The effects of autoclave and microwave heating on the protein and oil in soybean seeds were investigated under low moisture conditions. The nitrogen solubility index (NSI) decreased on heating. The reduction in the NSI was accompanied by an increase in the size and deformation of the oil bodies in the cellular tissue of soybean seeds. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that lipoxygenase was susceptible to heat denaturation, but 7S and 11S globulins were only partially denatured. The partial denaturation of the proteins was confirmed by Fourier transform infrared spectroscopy measurements. The ratio of oil to protein peaks increased with increasing heating, suggesting the exudation of oil to the surface or outside of oil bodies. Microwave heating is more efficient in changing the oil distribution in soybean seeds than autoclave heating. On the other hand, the degree of protein denaturation is lower after microwave heating.

Dietary low-fat soy milk powder retards diabetic nephropathy progression via inhibition of renal fibrosis and renal inflammation.

SCOPE: Diabetic nephropathy (DN) is a major cause of end-stage renal disease. Here, we examined the effect of long-term consumption of a low-fat soy milk powder (LFSMP) on the diabetic kidney structure and function. METHODS AND RESULTS: KKAy mice were fed a casein-, LFSMP-, or high-fat soy mixture powder (HFSMP)-based diet for 4 months. Plasma and urine were subjected to a biochemical assay every 2-4 wk. Renal morphology and protein expression were evaluated by histochemical staining and western blots. Although HFSMP-based diet showed no protective effect on DN. LFSMP-fed mice exhibited lower water intake, urine output, and urinary albumin, and glucose excretion. Furthermore, strong preservation of renal structural proteins and low urinary N-acetyl-beta-d-glucosaminidase activity were observed in LFSMP-fed mice, indicating alleviation of renal injury. LFSMP-fed mice showed a lesser degree of mesangial matrix expansion, of tubulointerstitial fibrosis, and of myofibroblast differentiation. Finally, milder renal inflammation was found in LFSMP-fed mice, as evidenced by a decrease in urinary monocyte chemoattractant protein- 1 excretion and lesser macrophage infiltration into the tubulointerstitium. CONCLUSION: The present data suggests that long-term consumption of LFSMP but not HFSMP retards DN progression via suppressing renal injury, myofibroblast differentiation, and renal macrophage infiltration in diabetic condition.

Decreases in serum triacylglycerol and visceral fat mediated by dietary soybean beta-conglycinin.

Soy protein isolate (SPI) is known to reduce the risk of heart disease by lowering serum cholesterol and triacylglycerol (TG) levels. Soybean beta-conglycinin, which is a component of SPI, might be the active ingredient that prevents and/or ameliorates lifestyle-related diseases, such as hyperlipidemia and obesity. This study aimed to determine the efficacy of soybean beta-conglycinin for lowering the human serum TG level and visceral fat. Randomized double-blind placebo-controlled designs were used to test the effect of dietary beta-conglycinin, which was taken in the form of candy. [Test 1]In order to examine the serum TG level, 138 volunteers aged 26 to 69 years with TG concentrations above 1.69 mmol/L participated in the study. The subjects were divided at random into two different groups: the test group only consumed the experimental candy containing beta-conglycinin and the placebo group only consumed the placebo candy containing casein. The test period consisted of a 2-wk pre-evaluation phase to screen the participants, a 12-wk consumption period and a 4-wk post-evaluation phase. The serum TG concentrations were significantly reduced in the test group, compared with the placebo group, after consuming the experimental candy. [Test 2]In order to measure visceral fat by means of CT scanning, 102 volunteers aged 26 to 69 years with body mass indices (BMI) between 25 and 30 participated in the study. The subjects were divided at random into two different groups as for Test 1. The test period consisted of a 2-wk pre-evaluation phase to screen the participants, a 20-wk consumption period and a 4-wk post-evaluation phase. A significant reduction in visceral fat only occurred in the beta-conglycinin group. This study showed that beta-conglycinin is an effective food ingredient that will be of use to reduce high serum TG concentrations and to prevent obesity.

Effects of soybean beta-conglycinin on body fat ratio and serum lipid levels in healthy volunteers of female university students.

The changes in body fat ratio and serum lipids induced by the ingestion of beta-conglycinin were examined in 41 healthy female university student volunteers. The trend of change in body fat ratio following the ingestion of beta-conglycinin differed between students with a baseline body fat ratio over 25% and those less than 25%. In the former group, the ingestion of beta-conglycinin suppressed the increase in body fat ratio. Moreover the six subjects who had a high total cholesterol level (5.72 mmol/L or higher) tended to have reduced levels of serum triglyceride, free fatty acid, total cholesterol and lipoprotein (a) after the ingestion of beta-conglycinin, although those levels did not change significantly. The number of subjects was only six, therefore it was inferred that significant changes were not observed. Thus, ingestion of soybean beta-conglycinin suppressed the increase in body fat ratio in individuals with a high baseline body fat ratio and reduced relatively high serum levels of lipids. Those results suggest that if soybean beta-conglycinin is ingested continuously (5 g daily), it will be effective in keeping body fat ratio and serum lipid levels normal and eliminating excessive lipids from the body.

Consumption of a structured triacylglycerol containing behenic and oleic acids increases fecal fat excretion in humans.

We examined the fecal fat excretion of mildly hypertriacylglycerolemic subjects who ingested soft cookies containing 1(3)-behenoyl-2,3(1)-dioleoyl-rac-glycerol (BOO) for 7 days. The subjects included 14 healthy men (average age; 44.9 ± 1.7) whose fasting plasma triacylglycerol level ranged from 150 to 250 mg/dL. Every day for 7 days, the subjects ate 5 soft cookies containing margarine with the BOO-rich experimental oil (BOO intake, 2.46 g/day). The placebo group ate soft cookies containing margarine without BOO. This study was a randomized double-blind, placebo-controlled, crossover study. Feces were collected for 3 days prior to the end of the treatment period, and fecal fat and fatty acid composition were determined. The fecal wet weight was significantly increased in BOO group compared with that in the placebo group. Moreover, fecal fat and fatty acid level were significantly higher in the BOO group than in the placebo group. There were no significant differences in the fecal fatty acid composition of the BOO and placebo groups. These results suggest that dietary BOO increases fecal excretion of dietary fat in humans. However, BOO does not increase the excretion of specific fatty acids; it increases the excretion of all fatty acids of dietary origin, which may lead to lower and delay intestinal absorption of dietary fat.

Effects of soy protein isolate feeding on severe kidney damage in DOCA salt-treated obese Zucker rats.

This study assessed the effects of soy protein isolate (SPI) on severe kidney damage in deoxycorticosterone acetate (DOCA) salt-treated obese Zucker rats. These rats underwent heminephrectomy and were fed either casein or SPI diet for 12 weeks. From weeks 8 to 10 of the experiment, kidney damage was induced by biweekly injection of 25 mg/kg DOCA and administration of 0.5% NaCl (w/v) ad libitum. Urinary protein and N-acetyl-ß-d-glucosaminidase excretions of SPI rats were much lower than those of casein rats at weeks 1 (p < 0.01) and 2 (p < 0.05) after DOCA treatment. Abnormal mineral excretions induced by DOCA treatment in casein rats were hardly detected in SPI rats. Severe atrophy of tubular epithelium and some flattened/detached renal tubules were also observed in casein rats, but not in SPI rats. These results indicate that consecutive treatment of SPI protects against renal dysfunction, particularly tubulointerstitial nephritis, in DOCA salt-treated obese Zucker rats.

Changes in lipid metabolism by soy beta-conglycinin-derived peptides in HepG2 cells.

In this study, HepG2 cells were treated with short peptides (7S-peptides) derived from highly purified soybean beta-conglycinin (7S), which was free from lipophilic protein, and the effect of the peptide treatment on lipid metabolism was determined. 7S-peptide treatment suppressed the secretion of apolipoprotein B-100 from HepG2 cells into the medium. The 7S-peptides also suppressed the incorporation of (3)H-glycerol and (14)C-acetate into triacylglyceride but not into major phospholipids, such as phosphatidylcholine and phosphatidylethanolamine. Additionally, the synthesis of cholesterol esters was dramatically decreased for 2 h after the addition of the 7S-peptides, whereas the synthesis of cholesterol remained unchanged by 4 h and increased by 8 h after the addition of the 7S-peptides. The cleaved nuclear form of SREBP-2 increased 8 h after the addition of the 7S peptides, suggesting a decrease in intracellular cholesterol levels. Analysis of changes in mRNA expression after 7S-peptide treatment suggested that the 7S-peptides lower the level of cholesterol in the endoplasmic reticulum, increase the mRNA of genes related to beta-oxidation of fatty acids, and increase the synthesis of cholesterol. From these results, it may be concluded that the peptides derived from 7S altered the lipid metabolism to decrease secretion of apolipoprotein B-100-containing lipoprotein from HepG2 cells.

Effects of soybean beta-conglycinin on hepatic lipid metabolism and fecal lipid excretion in normal adult rats.

beta-Conglycinin decreased blood triacylglycerol (TAG) levels in male Wistar adult rats. Liver mitochondrial carnitine palmitoyltransferase activity in the beta-conglycinin-fed group significantly increased as against the casein-fed group. Hepatic fatty acid synthase activity in the beta-conglycinin group significantly decreased as against that of the casein-fed group. Fecal fatty acid excretion in the beta-conglycinin group was significantly higher than in the casein group.