Preparation of corn ACE inhibitory peptide-ferrous chelate by dual-frequency ultrasound and its structure and stability analyses

In order to improve iron chelating ability and retain the activity of functional peptide, corn peptide was chelated with iron to form corn ACE inhibitory peptide-ferrous chelate (CP-Fe) treated by dual-frequency ultrasound. Furthermore, the chelating mechanism was revealed by analyzing various structural changes, and the stability was further evaluated. Under this study condition, the iron-binding capacity of corn ACE inhibitory peptide (CP) and chelate yield reached 66.39% and 82.87%, respectively. Ultrasound-treated CP exhibited a high iron chelating ability, meanwhile, chelation reaction had no significant effect on the ACE inhibition activity (82.21%) of the peptide. CP-Fe was formed by binding the peptides amino, carbonyl and carboxyl groups with Fe²⁺ demonstrated by Ultra-violet spectroscopy, Fourier transform infrared characterization, X-ray diffraction, energy dispersion spectrum, zeta potential, amino acid composition and other multi-angle analyses. Moreover, ultrasound-treated CP-Fe chelate exhibited porous surface and uniform nanoparticle shape. Furthermore, ultrasound-treated CP-Fe chelate exhibited an excellent stability towards various pH (retention rate ≥ 95.47% at pH 6–10), temperatures (retention rate ≥ 85.10% at 25–70 °C), and gastrointestinal digestion (retention rate 79.18%). Overall, ultrasound-treated CP-Fe chelate possessed high iron-chelating ability, ACE inhibition activity and stability. This study provides a novel synthesis method of the iron-chelating corn ACE inhibitory peptide, which is promising to be applied as iron supplements with high efficiency, bioactivity, and stability.     

Effects of ultrasound pretreatment with different frequencies and working modes on the enzymolysis and the structure characterization of rice protein

The effects of ultrasound pretreatment with different frequencies and working modes, including mono-frequency ultrasound (MFU), dual-frequency ultrasound (DFU) and tri-frequency ultrasound (TFU), on the degree of hydrolysis (DH) of rice protein (RP) and angiotensin-I-converting enzyme (ACE) inhibitory activity of RP hydrolysate were investigated. Ultraviolet–visible (UV) spectroscopy, fourier transform infrared (FTIR) spectroscopy, surface hydrophobicity and scanning electron microscopy (SEM) of RP pretreated with ultrasound were measured. The results showed that ultrasound pretreatment did not increase DH of RP significantly (p > 0.05). However, all the ultrasound pretreatment increased the ACE inhibitory activity of RP hydrolysate significantly (p < 0.05). The MFU of 20 kHz showed higher ACE inhibitory activity compared to that of other MFU. The ACE inhibitory activity of sequential DFU was higher than that of simultaneous with the same frequency combination. Sequential TFU of 20/35/50 kHz produced the highest increase in ACE inhibitory activity in contrast with other ultrasound frequencies and working modes. All the results under ultrasound pretreatment showed that ultrasound frequencies and working modes were of great effect on the ACE inhibitory activity of RP. The changes in UV–Vis spectra and surface hydrophobicity indicated the unfolding of protein and exposure of hydrophobic groups by ultrasound. The FTIR analysis showed that all the ultrasound pretreatment with different frequencies and working modes decreased α-helix, β-turn content and increased β-sheet, random coil content of RP. The SEM results indicated that ultrasound pretreatment resulted in the deformation of RP. In conclusion, the frequency selection of ultrasound pretreatment of RP is essential for the preparation of ACE inhibitory peptide.     

Dual-frequency multi-angle ultrasonic processing technology and its real-time monitoring on physicochemical properties of raw soymilk and soybean protein

To improve the soybean protein content (SPC), flavor and quality of soymilk, the effects of dual-frequency ultrasound at different angles (40 + 20 kHz 0°, 40 + 20 kHz 30°, 40 + 20 kHz 45°) on physicochemical properties and soybean protein (SP) structure of raw soymilk were mainly studied and compared with the conventional single-frequency (40 kHz, 20 kHz) ultrasound. Furthermore, the intensity of the ultrasonic field in real-time was monitored via the oscilloscope and spectrum analyzer. The results showed that 40 + 20 kHz 45° treatment significantly increased SPC. The ultrasonic field intensity of 40 + 20 kHz 0° treatment was the largest (8.727 × 10⁴ W/m²) and its distribution was the most uniform. The emulsifying stability of SP reached the peak value (233.80 min), and SP also had the largest particle size and excellent thermal stability. The protein solubility of 40 + 20 kHz 30° treatment attained peak value of 87.09%. 20 kHz treatment significantly affected the flavor of okara. The whiteness and brightness of raw soymilk treated with 40 kHz were the highest and the system was stable. Hence, the action mode of ultrasonic technology can be deeply explored and the feasibility for improving the quality of soymilk can be achieved.