Differential abilities of Korean soybean varieties to biosynthesize glyceollins by biotic and abiotic elicitors.

Glyceollins synthesized in soybeans that are exposed to biotic or abiotic stress have been reported to have health benefits. Considering that glyceollins are de novo synthesized from daidzein via several enzymatic steps and that isoflavone concentration widely varies among soybean varieties, the abilities of 60 soybean cultivars to synthesize glyceollins were compared under different elicitation conditions. Soybeans accumulated glyceollins differentially depending upon the cultivar when elicited with Aspergillus sojae. Contrary to our hypothesis that high isoflavone varieties may accumulate glyceollins more efficiently upon elicitation, glyceollin accumulation in response to fungal elicitation was not related with the concentration of either total isoflavones or daidzein in soybeans. Rather the glyceollin levels were significantly affected by soybean cultivar and most effectively increased by fungal infection. The data suggest that the selection of a strong fungal elicitor and a soybean cultivar with genotype that highly expresses the genes involved in glyceollin biosynthesis is essential for efficient glyceollin production.

Soybean-derived glyceollins induce apoptosis through ROS generation.

Glyceollins, which are synthesized from daidzein in soybeans infected with fungi, have been shown to have anti-fungal effects and antioxidant properties. However, the anti-proliferative mechanism of glyceollins against tumor cells is unknown. Glyceollin-induced apoptosis was evidenced by a decrease in cell viability and mitochondrial membrane potential, and an increase in early redistribution of plasma membrane phosphatidylserine, the sub G1 phase, and DNA fragmentation in hepa1c1c7 cells. Western blot analysis showed that treatment of the hepa1c1c7 cells with the glyceollins decreased the expression of pro-caspase-3, Bcl-2, and cell cycle-related proteins, but increased the expression of p21 and p27, and cytochrome C release into cytosol. At a concentration of 6 µg mL(-1) or higher, glyceollins significantly stimulated the production of reactive oxygen species (ROS), which appear to be responsible for the apoptotic activity of the compounds. Our present study demonstrated that the high dose of glyceollins possibly caused apoptosis in mouse hepatoma cells through the production of ROS, suggesting the potential to exploit glyceollins as anti-tumorigenic agents.

Comparative study of quality characteristics of Korean soy sauce made with soybeans germinated under dark and light conditions.

This study was conducted to evaluate the effects of germinating soybeans under dark and light conditions on the quality characteristics of Korean soy sauce made with germinated soybeans. The germination rate of soybeans germinated under dark conditions (GSD) was higher than that of soybeans germinated under light conditions (GSL), whereas the lengths of sprouts and relative weights of GSL did not differ from those of GSD. The L, a, b, and ΔT values of GSL were significantly lower than GSD. The color of GSD remained yellow, while GSL changed to a green color due to photosynthesis by chlorophyll. The total amino acid contents in soy sauce fermented with soybeans germinated under dark conditions (SSGD) and soy sauce fermented with soybeans germinated under light conditions (SSGL) were lower than in soy sauce fermented with non-germinated soybeans (SNGS). The levels of isoflavone content in SSGD and SSGL were significantly increased compared to the SNGS. In conclusion, the germination of soybeans under dark and light conditions is not only an increasing organoleptic preference, but also has implications for the health benefits of Korean soy sauce.

Effects of glyceollin I on vascular contraction in rat aorta.

The present study was undertaken to investigate the molecular mechanisms by which glyceollin I inhibits vascular contraction in rat aorta. Rat aortic rings were treated with either glyceollin I or vehicle when vascular contraction reached plateaus. We measured the activity of GTP-RhoA and Rho GTPase-activating protein (RhoGAP) and the phosphorylation level of the myosin light chain (MLC(20)), myosin phosphatase targeting subunit 1 (MYPT1), and PKC-potentiated inhibitory protein for heterotrimeric MLCP of 17 kDa (CPI17). Glyceollin I reduced vascular contraction whether endothelium is present or denuded. Glyceollin I reduced vascular contraction induced by NaF, U46619, phenylephrine, or PDBu. Blockers of K(+) channels did not affect the vasorelaxation induced by glyceollin I. Glyceollin I reduced activation of RhoA as well as phosphorylation level of MLC(20). Glyceollin I also reduced phosphorylation of MYPT1 and CPI17 induced by NaF but not PDBu. However, glyceollin I had no direct effect on RhoGAP activation in vitro. Glyceollin I reduced vascular contraction, at least in part, through inhibition of the RhoA/Rho-kinase signaling pathway.

Effects of light treatment on isoflavone content of germinated soybean seeds.

Our research objective was to increase isoflavone content in the germinated soybean seeds of four different varieties (Pungsannamulkong, Cheongjakong, Aga4, and Aga3) by optimizing light treatments (dark, greenhouse, fluorescent, incandescent, and ultraviolet lamps). The results demonstrated that the highest isoflavone content was produced from the Aga3 variety, which was developed by an interspecific cross between Eunhakong (Glycine max) and KLG10084 (G. soja) at the Plant Genetic Laboratory, Kyunpook National University. Aga3 is known to have one of the highest isoflavone content in the world at present. Our results recommend exposure of 7-day-old Aga3 sprouts to a combined light treatment of greenhouse lamps (12 h per day) and ultraviolet light (40 min per day) for maximum isoflavone production. Aga3 produced high levels of isoflavone because, in part, it contained very high isoflavone levels within the seed as compared with the other varieties. Under stress conditions, Aga3 could produce over 1.90 times more isoflavone than its seed content and 1.53 times more isoflavone than when grown under dark conditions.

Isoflavone attenuates vascular contraction through inhibition of the RhoA/Rho-kinase signaling pathway.

Isoflavones decrease blood pressure, improve lipid profiles, and restore vascular function. We hypothesized that isoflavone attenuates vascular contraction by inhibiting RhoA/Rho-kinase signaling pathway. Rat aortic rings were denuded of endothelium, mounted in organ baths, and contracted with 11,9 epoxymethano-prostaglandin F(2alpha) (U46619), a thromboxane A2 analog, or KCl 30 min after the pretreatment with genistein (4',5,7-trihydroxyisoflavone), daidzein (4',7-dihydroxyisoflavone), or vehicle. We determined the phosphorylation level of the myosin light chain (MLC(20)), myosin phosphatase-targeting subunit 1 (MYPT1), and protein kinase C-potentiated inhibitory protein for heterotrimeric myosin light-chain phosphatase of 17 kDa (CPI17) by means of the Western blot. We also measured the amount of GTP RhoA as a marker regarding RhoA activation. The cumulative additions of U46619 or KCl increased vascular tension in a concentration-dependent manner, which were inhibited by pretreatment with genistein or daidzein. Both U46619 (30 nM) and KCl (50 mM) increased MLC(20) phosphorylation levels, which were inhibited by genistein and daidzein. Furthermore, both genistein and daidzein decreased the amount of GTP RhoA activated by either U46619 or KCl. U46619 (30 nM) increased phosphorylation of the MYPT1(Thr855) and CPI17(Thr38), which were also inhibited by genistein or daidzein. However, neither genistein nor daidzein inhibited phorbol 12,13-dibutyrate-induced vascular contraction and CPI17 phosphorylation. In conclusion, isoflavone attenuates vascular contraction, at least in part, through inhibition of the RhoA/Rho-kinase signaling pathway.