The effects of processing and extraction conditions on content, profile, and stability of isoflavones in a soymilk system.

The effect of processing temperature and pH as well as enzyme-assisted extraction on the content and profile of isoflavones in a soymilk system was investigated. Isoflavone content in thermally treated soymilk at pH 7 and pH 9 was determined following a standard solvent extraction or an enzyme-assisted extraction protocol. Upon thermal processing, at both pH 7 and pH 9, significant interconversions were noted, indicated by the observed decrease in malonylglucosides with the concurrent increase in beta-glucosides. Enzyme-assisted extraction resulted in enhanced isoflavone extraction efficiency and revealed significant loss in total isoflavone content upon processing. This observation suggested that protein-isoflavone interactions, which are dependent on the protein structure and isoflavone form, affect isoflavone extractability, leading to underestimation of any loss that might have occurred in previously reported thermal studies. Accurate isoflavone measurements are essential to determine the processing conditions that result in the least loss of the biologically relevant isoflavone content.

Kinetic modeling of malonylgenistin and malonyldaidzin conversions under alkaline conditions and elevated temperatures.

The conversion and degradation of malonylglucosides were kinetically characterized under elevated pH/heat conditions. Malonylgenistin and malonyldaidzin were heated at 60, 80, and 100 degrees C and pH values of 8.5, 9, and 9.5. A simple kinetic model was developed, which adequately predicted the conversion and degradation reactions. The conversion and degradation rates increased as temperature and pH increased. The rates of conversion of both malonylglucosides into their respective beta-glucosides were comparable under all pH/heat treatments. However, at 100 degrees C, the rates of degradation of malonyldaidzin were approximately double those of malonylgenistin, under all pH treatments. When malonlydaidzin was heated at 100 degrees C and pH 9.5, degradation of the produced daidzin occurred. Therefore, an alternative kinetic model was developed to better predict the conversion and degradation of malonyldaidzin occurring at 100 degrees C and pH 9.5. The models developed provide soy food manufacturers with guidelines for better control of the profile and level of isoflavones..

Heat and pH effects on the conjugated forms of genistin and daidzin isoflavones.

Isoflavones occur primarily as glycosides (namely, malonyl-, acetyl-, and non-conjugated beta-glycosides) and a small percentage as the bioactive aglycon. The different chemical structures of isoflavones can dictate their stability during processing. Therefore, our objective was to determine the effects of pH and thermal treatments on conjugated isoflavones with regard to interconversions and loss. Conjugated daidzin and genistin were heated at 25, 80, and 100 degrees C under neutral, acidic, and basic conditions. Changes in isoflavone derivatives were monitored using high-performance liquid chromatography. Along with interconversions, considerable loss in total known isoflavone derivatives was noted for each isoflavone, especially under elevated pH and temperature. The malonylglycosides showed more stability than acetylglycosides, especially under acidic conditions. Overall, loss in isoflavone derivatives was significantly higher for daidzin than for genistin glycoside forms. Our results highlighted the significance of chemical structure with regard to stability, which is a key factor in determining soy processing conditions.

FTIR determination of ligand-induced secondary and tertiary structural changes in bovine plasminogen.

Human plasminogen undergoes a large tertiary structural change in the presence of lysine derivatives (e.g. epsilon-amino caproic acid, EACA). This change facilitates human plasminogen activation by human plasminogen activators, resulting in elevated blood plasmin levels. It is hypothesized that this structure-function relationship is similar for bovine plasminogen. The objectives of this study were to investigate the effect of the ligand EACA on the secondary structure of plasminogen (bovine, human, and rabbit) and the tertiary structure of bovine plasminogen using Fourier-transform infrared spectroscopy (FTIR). Spectra of plasminogen, EACA, and a mixture of plasminogen and EACA in water and deuterium were collected using FTIR. Fourier-self deconvoluted spectra in the amide I region (1700-1600 cm(-1)) were used to detect changes in secondary structure of plasminogen after EACA addition. Change in bovine plasminogen tertiary structure was determined by comparing ratios of amide II (1600-1500 cm(-1)) to amide I bond intensities over time for samples in deuterium. No differences in secondary structure were observed for any plasminogen in the presence of EACA; however, addition of EACA significantly changed tertiary structure of bovine plasminogen. This tertiary structural change indicates a transition from a folded to an unfolded state, which could be more easily converted to plasmin. These results are consistent with reported human plasminogen studies using neutron scattering (tertiary structure) and circular dichroism (secondary structure) methods.