Haile Ma

Continuous and pulsed ultrasound-assisted extractions of antioxidants from pomegranate peel

There is a great demand for developing efficient extraction methods in order to reduce extraction time and increase the yield and activity of functional antioxidants. The yields, activities, and extraction kinetics of antioxidants from the dry peel of pomegranate marc were studied using ultrasound-assisted extraction in continuous and pulsed modes and the results were compared with conventional extraction (CE) at a temperature of 25±2 °C and water/peel ratio of 50/1, w/w. The studied factors were intensity level and treatment time for continuous ultrasound-assisted extraction (CUAE), and intensity level, number of pulse repetition, and pulse duration and interval for pulsed ultrasound-assisted extraction (PUAE). The results showed that all factors significantly affected the antioxidant yield, but only treatment time had a significant effect on the antioxidant activity. Compared to CE, PUAE at an intensity level of 59.2 W/cm(2), and a pulse duration of 5s and a resting interval of 5s increased the antioxidant yield by 22% and reduced the extraction time by 87%. Similarly, CUAE at the same intensity level increased the antioxidant yield by 24% and reduced the extraction time by 90%. Since PUAE had 50% energy saving compared to CUAE, we recommend using PUAE for the extraction with antioxidant yield of 14.5% and DPPH scavenging activity of 5.8 g/g. A second-order kinetic model was successfully developed for describing the mechanism of ultrasound-assisted extractions under PUAE and CUAE. This research clearly demonstrated the superiority of PUAE for producing antioxidants from peel of pomegranate marc.

Effect of energy-gathered ultrasound on Alcalase

This research was to explore the mechanism of ultrasonic impact on protease activity. The effects of energy-gathered ultrasound on the activity, kinetics, thermodynamics and molecular structure of Alcalase were investigated with the aid of the chemical reaction kinetics model, Arrhenius equation, Eyring transition state theory, fluorescence spectroscopy and circular dichroism (CD) spectroscopy. Results showed that ultrasound had effect on the activity of Alcalase. The highest Alcalase activity was achieved when the sample was treated with energy-gathered ultrasound at 80 W for 4 min, under which the enzyme activity was increased by 5.8% over the control. After the treatment, thermodynamics parameters Ea, ΔH, ΔS and ΔG were reduced by 70.0%, 75.8%, 34.0% and 1.3%, respectively. Besides, fluorescence and CD spectra revealed that the ultrasonic treatment had increased the number of tryptophan on Alcalase surface slightly, increased number of α-helix by 5.2%, and reduced the number of random coil by 13.6%.

Quantitative determination of major polyphenol constituents in pomegranate products

The anti-oxidant content and potential health benefits associated with consuming pomegranate and pomegranate-containing products has lead to increased consumer demand for this crop resulting in it becoming a high value crop. The potential health benefits and high anti-oxidant content of this fruit is attributed to the polyphenolic compounds it contains, including the ubiquitous phenolic acids, gallic acid and ellagic acid, and punicalagin A and punicalagin B, two polyphenolics unique to this fruit. A rapid HPLC-UV method targeting these four metabolites requiring minimal sample cleanup and offering runtimes half as long as existing methods was established. Within day and inter-day run-to-run variability for the four metabolites ranged from 1.9% to 6.6% and 5.3% to 11.4%, respectively. Spike recovery percentages for gallic acid, punicalagin A, punicalagin B and ellagic acid were found to be 98.5%, 92.4%, 95.5%, and 96.5%, respectively. This method was applied to the evaluation of various pomegranate products, including commercial drinks, handmade juice, and marc extracts. This method may be readily used to verify the presence of pomegranate metabolites in juices, extracts, and other products.

Enzymolysis kinetics and activities of ACE inhibitory peptides from wheat germ protein prepared with SFP ultrasound-assisted processing

There is a great demand for developing efficient enzymolysis methods in order to increase the enzymolysis efficiencies and activities of angiotensin converting enzyme (ACE) inhibitory peptides from wheat germ protein. The enzymolysis kinetics, ACE inhibitory activity of peptide and conversion rate of protein were studied using sweep frequency and pulsed (SFP) ultrasound-assisted enzymolysis and the results were compared with traditional enzymolysis. The studied factors were enzymolysis time and substrate concentration. By considering the activity of ACE inhibitory peptide and operation cost, the recommended conditions of SFP ultrasound-assisted enzymolysis were enzymolysis time of 120 min and substrate concentration of 24.0 g/L, which gave high conversion rates of protein (60.7%) and ACE inhibitory activity of peptide (65.9%). Compared to traditional enzymolysis, SFP ultrasound-assisted enzymolysis significantly increased the initial reaction rate (V) by 60.0% at substrate concentration of 24.0 g/L, increased the apparent breakdown rate constant (k(A)) by 66.7%, decreased the apparent constant (K(M)) by 6.9%, and raised the conversion rate of protein by 35.5% and ACE inhibitory activity of peptides by 35.6% under the recommended conditions. It has been concluded that SFP ultrasound can remarkably raise the enzymolysis efficiency and activity of ACE inhibitory peptides from wheat germ protein.

Pretreatment of defatted wheat germ proteins (by-products of flour mill industry) using ultrasonic horn and bath reactors: Effect on structure and preparation of ACE-inhibitory peptides

The ultrasonic horn and bath reactors were compared based on production of angiotensin-converting-enzyme (ACE) inhibitory peptides from defatted wheat germ proteins (DWGP). The DWGP was sonicated before hydrolysis by Alcalase. Degree of hydrolysis, ACE-inhibitory activity, surface hydrophobicity, fluorescence intensity, free sulfhydryl (SH), and disulfide bond (SS) were determined. The highest ACE-inhibitory activity of DWGP hydrolysate was obtained at power intensity of 191.1 W/cm(2) for 10 min in the ultrasonic horn reactor. The fixed frequency of 33 kHz and the sweep frequency of 40±2 kHz resulted in the maximum ACE-inhibitory activity. The combined irradiation of dual fixed frequency (24/68 kHz) produced significant increase in ACE-inhibitory activity compared with single frequency (33 kHz). The ultrasonic probe resulted in significant higher ACE-inhibitory activity compared with ultrasonic bath operating at single or dual fixed and sweep frequencies. The changes in conformation of the DWGP due to sonication were confirmed by the changes in fluorescence intensity, surface hydrophobicity, SHf and SS contents and they were found in conformity with the ACE-inhibitory activity in case of the ultrasonic horn reactor but not in bath reactor.

Extract of Phenolics From Pomegranate Peels

The effects of different solvents, temperature conditions, solvent-solid ratios and particle sizes on solid-solvent extraction of the total phenolics, proanthocyanidins and flavonoids herein also referred to as antioxidant from pomegran- ate marc peel (PMP) was studied. Water, methanol, ethanol, acetone, and ethyl acetate extraction efficiencies at extraction times of 0.17 to 10 min, extraction temperatures of 25 to 95°C, ratios of solvent/solid of 5:1 to 50:1 and particle sizes of 10 to 40 mesh were evaluated. At 40 °C, solvent/solid ratio of 15:1, extraction time of 240 min and particle size of 40 mesh, methanol gave the highest extract yield of the total phenolics (8.26%), followed by water (5.90%), ethanol (1.55%), acetone (0.37%), and ethyl acetate (0.18%), respectively. However, at an extraction temperature of 95°C, the total phe- nolics extract yield with water was 11.15% for particle size of 40 mesh, solvent/solid ratio of 15:1, and extraction time of 2 min. Despite the lowest extract yield at extraction temperature of 40 °C, solvent/solid ratio of 15:1, extraction time of 240 min and particle size of 40 mesh, ethyl acetate extraction gave the highest content of the total phenolics (20.24%), proanthocyanidins (2.65%) and flavonoids (3.92%) in the extract. The DPPH antioxidant activity of extracts had a linear relationship with the total phenolics content in the extracts (R 2 =0.9779). This study revealed that water extraction, which has the economic and safety merits, can be used as an environmentally friendly method for producing antioxidants from the PMP.

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 β-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 β-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.

Effect of extraction media on preliminary characterizations and antioxidant activities of Phellinus linteus polysaccharides

Three partially purified polysaccharides were extracted from Phellinus linteus mycelia using hot water, 1% (NH4)2C2O4, and 1.25M NaOH/0.05% NaBH4, and the extracts were named PL-W, PL-A, and, PL-N respectively. PL-N mainly comprised xylose and arabinose with a high molecular weight (Mw) and the highest carbohydrate and uronic acid contents. PL-W and PL-A were mainly composed of glucose with high and low Mw fractions in various ratios. All three polysaccharides existed as compact coils in aqueous solutions and exhibited strong scavenging capacity and antioxidant activities in a concentration-dependent manner. The polysaccharides also had high uronic acid and carbohydrate contents and strong antioxidant activities. The Mws, monosaccharide compositions, and chemical structures of the polysaccharides also affected their antioxidant activities. PL-A and PL-N had better antioxidant activities and could thus be developed as potential natural antioxidant agents for applications in food additives and biomedical industries.

Effects of ultrasound and ultrasound assisted alkaline pretreatments on the enzymolysis and structural characteristics of rice protein.

The objectives of this study were to investigate the effects of multi-frequency energy-gathered ultrasound (MFEGU) and MFEGU assisted alkaline pretreatments on the enzymolysis and the mechanism of two pretreatments accelerating the rice protein (RP) proteolysis process. The results showed that MFEGU and MFEGU assisted alkaline pretreatments improved significantly (P<0.05) the degree of hydrolysis (DH) and the protein elution amount of RP. Furthermore under the same DH conditions, ultrasound and ultrasound assisted alkaline pretreatments were more save the enzymolysis time than the unpretreatment. The changes in UV-vis spectra, fluorescence emission spectra indicated unfolding and destruction of RP by MFEGU and MFEGU assisted alkaline pretreatments. The circular dichroism analysis showed that both pretreatments decreased α-helix but increased β-sheet and random coil of RP. Amino acid composition revealed that MFEGU and MFEGU assisted alkaline pretreatments could increase the protein elution amount and the ratio of hydrophobic amino acids. Atomic force microscopy (AFM) indicated that both pretreatments destroyed the microstructures and reduced the particle size of RP. Therefore, MFEGU and MFEGU assisted alkaline pretreatments are beneficial to improving the degree of hydrolysis due to its sonochemistry effect on the molecular conformation as well as on the microstructure of protein.

Modeling the QSAR of ACE-Inhibitory Peptides with ANN and Its Applied Illustration.

A quantitative structure-activity relationship (QSAR) model of angiotensin-converting enzyme- (ACE-) inhibitory peptides was built with an artificial neural network (ANN) approach based on structural or activity data of 58 dipeptides (including peptide activity, hydrophilic amino acids content, three-dimensional shape, size, and electrical parameters), the overall correlation coefficient of the predicted versus actual data points is R = 0.928, and the model was applied in ACE-inhibitory peptides preparation from defatted wheat germ protein (DWGP). According to the QSAR model, the C-terminal of the peptide was found to have principal importance on ACE-inhibitory activity, that is, if the C-terminal is hydrophobic amino acid, the peptides ACE-inhibitory activity will be high, and proteins which contain abundant hydrophobic amino acids are suitable to produce ACE-inhibitory peptides. According to the model, DWGP is a good protein material to produce ACE-inhibitory peptides because it contains 42.84% of hydrophobic amino acids, and structural information analysis from the QSAR model showed that proteases of Alcalase and Neutrase were suitable candidates for ACE-inhibitory peptides preparation from DWGP. Considering higher DH and similar ACE-inhibitory activity of hydrolysate compared with Neutrase, Alcalase was finally selected through experimental study.