The milk macromolecular peptide: preparation and evaluation of antihypertensive activity in rats.

In order to avoid the discomfort of digesting milk protein and make full use of the gastrointestinal digestive function, a milk macromolecular peptide was prepared with ACE inhibitory activity after gastrointestinal digestion as the index. Then, the antihypertensive activity of the milk macromolecular peptide in rats was evaluated. The results showed that the ACE inhibitory activity of hydrolysate after simulated gastrointestinal digestion was the highest in which the degree of hydrolysis was 17% and the ACE inhibitory activity reached 78.48%. The optimum enzymatic parameters were obtained with the protein concentration of 3.4%, enzymatic temperature of 55 °C, pH value of 7.0, and 7% enzyme amount using neutral protease. Under these conditions, the ACE inhibitory activity of hydrolysate after simulated gastrointestinal digestion was as high as 89.49%. Compared to traditional peptides, the ACE inhibitory activity of the macromolecular peptide increased after simulated gastrointestinal digestion. After 4 weeks of gavage tests of the milk macromolecular peptide, the blood pressure of rats with spontaneous hypertension dropped to below 140 mmHg, with a decrease of 60 mmHg. The antihypertensive effect of the milk macromolecular peptide was similar to that of captopril. The milk macromolecular peptide decreased the blood pressure of spontaneously hypertensive rats by inhibiting the ACE activity of the lung, kidney and arteries, and regulating the content of the signal factors of endothelin, NO, angiotensin II and renin in serum indicators. These results indicated that the macromolecular ACE inhibitory peptide with an antihypertensive effect could be prepared by the enzymatic hydrolysis of milk protein to realize the full utilization of protein resources.

Ultrasound-assisted preparation of ACE inhibitory peptide from milk protein and establishment of its in-situ real-time infrared monitoring model.

A scheme for preparing milk ACE inhibitory peptides by in vitro proteolysis and simulated gastrointestinal digestion was constructed. The ultrasonic assisted pretreatment was used to improve the enzymolysis of milk protein. The in-situ real-time infrared was used to establish monitoring model of enzymatic process. Results showed that under the conditions of single frequency 28 kHz, ultrasound time 40 min, ultrasound power density 20 W/L, milk protein concentration 34 g/L, batch ratio 2:4 and initial temperature 30 °C, the ACE inhibition rate of gastric digestion of enzymatic hydrolysate reached 67.20%, which was 22.87% higher than that of non-ultrasound samples. The results of secondary structure studies of proteins showed that after the ultrasonic treatment, the content of α-helix and ß-corner reduced, and the content of ß-folding and random coil increased. Compared with the control group, the ultrasonic treatment increased surface hydrophobicity and the content of SH while reduced the content of SS in milk protein, thus improving the ACE inhibitory activity of enzymatic hydrolysates. Furthermore, three quantitative prediction models of PLS, iPLS and Si-PLS for ACE inhibition rate of milk protease hydrolysates were established. And all these three different in-situ real-time prediction models had good predictive effect on the ACE inhibition rate of milk proteolysis products and gastrointestinal simulated digestion products.

Effects and mechanism of ultrasound pretreatment on rapeseed protein enzymolysis.

BACKGROUND: The disadvantages which stem from the use of traditional enzymolysis of protein has necessitated the need to employ sweeping frequency and pulsed ultrasound (SFPU) in the pretreatment of rapeseed protein prior to proteolysis in order to bring about improvement in enzymolysis efficiency. Further, in order to determine the mechanism of ultrasound-accelerated enzymolysis of RP, the effects of SFPU on the kinetics, thermodynamics, molecular conformation and microstructure of RP were investigated. RESULTS: Kinetic studies showed that SFPU pretreatment on RP improved enzymolysis by decreasing the apparent constant KM significantly (P < 0.05) by 32.8% and reducing the thermodynamic parameters Ea , ΔH and ΔS by 16.6%, 17.7% and 9.2% respectively. Fluorescence spectra revealed that SFPU pretreatment induced molecular unfolding, causing more hydrophobic groups and regions inside the molecules to be exposed to the outside. Circular dichroism analysis indicated that SFPU pretreatment decreased the α-helix content by 16.1% and increased the random coil content by 3.6%. In addition, scanning electron microscopy showed that SFPU pretreatment increased the specific surface area of RP. CONCLUSION: Ultrasound pretreatment is an efficient method in RP proteolysis to produce peptides through its impact on the molecular conformation and microstructure of proteins.

Effects of multi-frequency power ultrasound on the enzymolysis of corn gluten meal: Kinetics and thermodynamics study.

The effects of multi-frequency power ultrasound (MPU) pretreatment on the kinetics and thermodynamics of corn gluten meal (CGM) were investigated in this research. The apparent constant (KM), apparent break-down rate constant (kA), reaction rate constants (k), energy of activation (Ea), enthalpy of activation (ΔH), entropy of activation (ΔS) and Gibbs free energy of activation (ΔG) were determined by means of the Michaelis-Menten equation, first-order kinetics model, Arrhenius equation and transition state theory, respectively. The results showed that MPU pretreatment can accelerate the enzymolysis of CGM under different enzymolysis conditions, viz. substrate concentration, enzyme concentration, pH, and temperature. Kinetics analysis revealed that MPU pretreatment decreased the KM value by 26.1% and increased the kA value by 7.3%, indicating ultrasound pretreatment increased the affinity between enzyme and substrate. In addition, the values of k for ultrasound pretreatment were increased by 84.8%, 41.9%, 28.9%, and 18.8% at the temperature of 293, 303, 313 and 323 K, respectively. For the thermodynamic parameters, ultrasound decreased Ea, ΔH and ΔS by 23.0%, 24.3% and 25.3%, respectively, but ultrasound had little change in ΔG value in the temperature range of 293-323 K. In conclusion, MPU pretreatment could remarkably enhance the enzymolysis of CGM, and this method can be applied to protein proteolysis industry to produce peptides.

Antioxidant peptides from corn gluten meal: Orthogonal design evaluation.

Protamex catalyzed corn gluten meal (CGM) hydrolysis peptides (CHP) were prepared. Orthogonal design L16 (4(5)) was used to optimize processing variables of CGM concentration, CGM heat pretreatment (121 °C) time, and enzymolysis pH, temperature, and time. Degree of hydrolysis (DH), undigested residue ratio, molecular weight (MW) distribution and DPPH radical inhibition were selected as analysis indicators. Optimum variables were CGM concentration of 18%, heat pretreatment time of 40 min, and enzymolysis pH, temperature and time of 7.5, 55 °C and 24h, respectively. Verification test showed that CHP IC50 for scavenging hydroxyl radical was the best and then followed by reducing power. Oligopeptides improved after hydrolysis at the expense of di- and tripeptides, suggesting formation of soluble aggregates from low MW peptides. The increase in the DH, oligopeptides, Alanyl-Tyrosine, and antioxidant free amino acids coincided with the improvement in the antioxidant activity of CHP.

Effects of multi-frequency power ultrasound on the enzymolysis and structural characteristics of corn gluten meal.

The aim of this study was to investigate the effect of multi-frequency power ultrasound (sweeping frequency and pulsed ultrasound (SFPU) and sequential dual frequency ultrasound (SDFU)) on the enzymolysis of corn gluten meal (CGM) and on the structures of the major protein fractions (zein, glutelin) of CGM. The results showed that multi-frequency power ultrasound pretreatments improved significantly (P<0.05) the degree of hydrolysis and conversion rate of CGM. The changes in UV-Vis spectra, fluorescence emission spectra, surface hydrophobicity (H0), and the content of SH and SS groups indicated unfolding of zein and glutelin by ultrasound. The circular dichroism analysis showed that both pretreatments decreased α-helix and increased ß-sheet of glutelin. The SFPU pretreatment had little impact on the secondary structure of zein, while the SDFU increased the α-helix and decreased the ß-sheet remarkably. Scanning electron microscope indicated that both pretreatments destroyed the microstructures of glutelin and CGM, reduced the particle size of zein despite that the SDFU induced aggregation was observed. In conclusion, multi-frequency power ultrasound pretreatment is an efficient method in protein proteolysis due to its sonochemistry effect on the molecular conformation as well as on the microstructure of protein.

Examining of athermal effects in microwave-induced glucose/glycine reaction and degradation of polysaccharide from Porphyra yezoensis.

Many reports claim the existence of athermal effects in microwave-induced reactions, and this challenge the assumption that the thermal effect (heating) is the sole factor in microwave heating. Therefore, microwave-induced Maillard reaction of d-glucose/glycine and degradation of polysaccharide from Porphyra yezoensis (PSPY) were investigated. Browning reactions were monitored by measuring heating rate, UV-absorbance and brown color, UV-vis and synchronous fluorescence spectra, GC/MS analysis and intrinsic viscosity of degradation. Heating of d-glucose/glycine solution produced brown compounds which were detected at A420, and the intermediate products, 2-acetylfuran and 5-methylfurfural, whose fluorescence intensity evidenced their formation. Maximum emission of synchronous fluorescence spectra of samples were at 430-440 nm and 370-390 nm. Both microwave and water bath heating did not cause any compositional changes in the Maillard reaction products. All data failed to show any significant athermal effects of compositional changes in the Maillard reaction products. It can be inferred that some of the reports suggesting the existence of athermal effects, which could ascribe to the different set-up obtained in not well temperature controlled microwave heating systems.