Purification and identification of lipolysis-stimulating peptides derived from enzymatic hydrolysis of soy protein.

The aim of this study was to purify and identify lipolysis-stimulating peptides derived from Flavourzyme®-soy protein isolate (SPI) hydrolysate (F-SPIH). Glycerol release was employed as a marker for lipolysis in 3T3-L1 adipocytes. A higher glycerol release represents a better lipolysis-stimulating activity. The peptide fraction with highest glycerol release obtained from F-SPIH fractionated by sequential ultrafiltration membranes was further purified using gel filtration chromatography and two steps of reverse-phase high-performance liquid chromatography. The peptides were identified using liquid chromatography-tandem mass spectrometry (LC/MS/MS). Three lipolysis-stimulating peptides were obtained, and the amino acid sequences were ILL, LLL and VHVV, respectively. The in vitro effect of gastrointestinal proteases on lipolysis-stimulating activity of synthetic ILL, LLL and VHVV, respectively, was also investigated. The result suggested that the gastrointestinal protease did not affect lipolysis-stimulating activity of the three novel peptides, which reveals their potential to act as anti-obesity ingredients.

Effect of water-to-bean ratio on the contents and compositions of isoflavones in tofu.

The present study investigated the changes of the bioavailable isoflavones, including daidzin, genistin, daidzein, and genistein, during the making of tofu. The amount of extracted daidzin and genistin in soy milk increased with increasing water-to-bean ratios from 5 to 9 and reached the maximum level at the ratios of 9-11. On the other hand, the amount of extracted free isoflavones (daidzein and genistein) was not affected by the water-to-bean ratio at the range of 5-11, and their extracted amounts in soy milk were 2-4-fold those in raw soybean. It is suggested that these free isoflavones are mainly derived from daidzin, genistin, malonyldaidzin, and malonylgenistin through enzymatic hydrolysis during the making of soy milk. Tofu made with water-to-bean ratios of 9:1 and 10:1 had the maximal retentions of daidzin and genistin, which were due to the fine homogeneous network microstructure that is supposed to be more effectively retained through hydrophilic interaction with protein. On the contrary, the retained amount of free isoflavones decreased significantly as the water-to-bean ratio increased from 7 to 11, due to their weakening hydrophobic interaction with protein. In this study it was found that the homogeneous microstructure of tofu improved the retention of hydrophilic daidzin and genistin and that the increased amount of drained water does not significantly reduce their retention in the final tofu products as generally imagined.

Effect of calcium sulfate concentration in soymilk on the microstructure of firm tofu and the protein constitutions in tofu whey.

The effects of calcium sulfate concentration in soymilk on the microstructure of tofu and the constitutions of protein in tofu whey were investigated. The firm tofu made with 0.4% CaSO(4).2H(2)O was found to be most uniform and continuous in the microstructure. This tofu gave the maximal tofu yield, maximal solid and protein recoveries, and the best water retention ability. The results of gel electrophoresis and the ratio of amino nitrogen to total organic nitrogen indicated that the low molecular weight proteins in tofu whey were at their lowest when the tofu was made with 0.4% CaSO(4).2H(2)O. The SEM observations suggested that the tofu made with 0.4% CaSO(4).2H(2)O has the most uniform and homogeneous microstructure and, consequently, can most efficiently retain soybean proteins and water in the tofu gel.