Preparation Process Optimization of Peptides from Agaricus blazei Murrill, and Comparison of Their Antioxidant and Immune-Enhancing Activities Separated by Ultrafiltration Membrane Technology.

Agaricus blazei murrill (ABM), a large fungus, is reported to have extensive biological activities but the antioxidant and immune-regulatory capacities have been less studied and the components responsible for the functions are unclear. This study prepared ABM peptides (ABMP) using ultrasound-assisted enzymatic extraction (UAEE) strategy and cascade ultrafiltration (UF) membrane technology. The UAEE extraction conditions were optimized using response surface methodology (RSM) with four factors and three levels to achieve the maximum ABMP yield (34.03%); the optimal conditions were an enzyme amount of 4%, ratio of ABM to water of 1:30, ultrasonic power of 360 W, and ultrasonic time of 30 min. Four ABMP fractions were obtained after UF with different pore size and their antioxidant and immune-regulatory abilities were evaluated and compared. The results showed that they could effectively scavenge DPPH, hydroxyl, and ABTS radicals, especially for ABMP-2; the scavenging rate of the above radicals were 79.31%, 63.60%, and 96.08%, respectively. In addition, four ABMP fractions also activated macrophage activity through strengthening phagocytosis and the production of NO, IL-6, IL-1ß, and TNF-α in a dose-dependent manner. Notably, the ABMP-2 fraction with a MW of 3-5 kDa and peptide purity of 82.88% was found to have the best effect, showing the maximum phagocytosis (189.37%) as well as NO (7.98 µM), IL-6 (195.05 pg/mL), IL-1ß (876.15 pg/mL), and TNF-α (1620 pg/mL) secretion at a treatment concentration of 150 µg/mL. The findings indicated that the ABMP, especially for the separate ABMP-2, could be used as dietary supplements and have the potential to be exploited as immune-enhancing agents.

Effects of ultrasonic pre-treatment on physicochemical properties of proteins extracted from cold-pressed sesame cake.

Sesame is an oil crop with high nutritional value. Protein is one of the main ingredients of sesame, however research on protein of cold-pressed sesame cake is limited. This study aimed to investigate the effects of ultrasonic pre-treatment (UPT) on physicochemical properties of proteins (yield, solubility, amino acid composition, surface properties, structural and thermal stability) extracted from the cold-pressed sesame cake, after removing lignans by ultrasonic-assisted extraction. By comparison, the extraction yield of protein was significantly (p ＜ 0.05) increased from 22.24% (without UPT) to 25.95% (with UPT), while the purity (54.08% without UPT, 55.43% with UPT), total amount of essential amino acids (22.48% without UPT, 23.10% with UPT) and non-essential amino acids (37.48% without UPT, 36.54% with UPT) were not significantly influenced. Besides, UPT slightly reduced the solubility, foaming capacity and stability (FC and FS) of protein. In addition, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR) and thermal stability (TG) analysis demonstrated that UPT could disorder and loose protein molecular structure, resulting in the change of morphology, secondary structure and thermal stability. In conclusion, this study provides a way for the separation and future application of sesame cake protein. UPT is a good option to remove the lignans from sesame cake proteins.

Effects of Chinese Dietary Pattern of Fat Content, n-6/n-3 Polyunsaturated Fatty Acid Ratio, and Cholesterol Content on Lipid Profile in Rats.

This study aims to investigate the effect of Chinese diet pattern of fat content (30% or 36.06%), n-6/n-3 polyunsaturated fatty acid (PUFA) ratio (5 : 1 or 9 : 1), and cholesterol content (0.04 or 0.057 g/kg total diet) on lipid profile using a rat model. Results showed that rats' body weights (BWs) were controlled by the simultaneous intakes of cholesterol level of 0.04 g/kg total diet and n-6/n-3 ratio of 5 : 1. In addition, under high-fat diet, increased cholesterol feeding led to increased total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) levels and decreased triacylglycerols (TG) in rats' plasma. However, high density lipoprotein cholesterol (HDL-C) level and the ratios of HDL-C/LDL-C and HDL-C/TC in rats' plasma increased in response to simultaneous intakes of low n-6/n-3 ratio (5 : 1) and cholesterol (0.04 g/kg total diet) even under high-fat diet. Moreover, as the n-6/n-3 PUFA ratio in the diet decreased, the proportion of n-3 PUFAs increased in plasma, liver, and muscle and resulted in the decrease of n-6/n-3 PUFA ratio.

Methionine sulfone-containing orbitides, good indicators to evaluate oxidation process of flaxseed oil.

This study aimed to explore the possibility of using methionine sulfone (Msn)-containing orbitides as indicators to evaluate the oxidation process of flaxseed oils. Results showed that after 4 days' heating, oxidation values slightly increased (p > .05) with significant decrease in methionine (Met)-containing peptides (p < .05) instead of γ-tocopherol (p > .05). However, as oxidation time continues increasing, oxidation values significantly increased (p < .05) with significant reduction of γ-tocopherol (p < .05). It demonstrated that Met-containing peptides were more readily oxidized compared with γ-tocopherol and showed certain antioxidant activity. Besides, high logarithmic correlations were found between oxidation values and Msn-containing orbitides (0.94-1.00), such as between total carbonyl compounds and orbitide [1-8-NαC],[1-MetO2]-CLE (64.95 lnx - 52.14, R2 = 0.99, Dingya23 oil). Therefore, in comparison with common oxidation indices, Msn-containing orbitides may be better indicators for evaluating the oxidation process of flaxseed oil with superior separation efficiency, specific information and high stability.

Esterification of Quercetin Increases Its Transport Across Human Caco-2 Cells.

Plant polyphenols showed useful biochemical characteristics in vitro; however, the assessments of their clinical applications in vivo are restricted by their limited bioavailability due to their strong resistance to 1st-pass effects during absorption. In order to improve the bioavailability of quercetin (QU), the ester derivative of QU (3,3',4',5,7-pentahydroxy flavones, TAQU) was synthesized, followed by examining the oil-water partition coefficient as well as the transport mechanisms of QU and its ester derivative (TAQU) using human Caco-2 cells. The transport characteristics of QU and TAQU transport under different conditions (different concentrations, time, pH, temperature, tight junctions, and potential transporters) were systematically investigated. Results showed that QU had a lower permeability coefficient (2.82 × 10(-6) cm/s) for apical-to-basolateral (AP-BL) transport over 5 to 50 µM, whereas the transport rate for AP to BL flux of TAQU (5.23 × 10(-6) cm/s) was significantly greater than that of QU. Paracellular pathways were not involved during the transport of both QU and TAQU. QU was poorly absorbed by active transport, whereas TAQU was mostly absorbed by passive diffusion. Efflux transporters, P-glycoproteins, multidrug resistance proteins were proven to participate in the transport process of QU, but not in that of TAQU. These results suggested that improving the lipophicity of QU by esterification could increase the transport of QU across Caco-2 cells.

Lipozyme RM IM-catalyzed acidolysis of Cinnamomum camphora seed oil with oleic acid to produce human milk fat substitutes enriched in medium-chain fatty acids.

In the present study, a human milk fat substitute (HMFS) enriched in medium-chain fatty acids (MCFAs) was synthesized through acidolysis reaction from Cinnamomum camphora seed oil (CCSO) with oleic acid in a solvent-free system. A commercial immobilized lipase, Lipozyme RM IM, from Rhizomucor miehei, was facilitated as a biocatalyst. Effects of different reaction conditions, including substrate molar ratio, enzyme concentration, reaction temperature, and reaction time were investigated using response surface methodology (RSM) to obtain the optimal oleic acid incorporation. After optimization, results showed that the maximal incorporation of oleic acid into HMFS was 59.68%. Compared with CCSO, medium-chain fatty acids at the sn-2 position of HMFS accounted for >70%, whereas oleic acid was occupied predominantly at the sn-1,3 position (78.69%). Meanwhile, triacylglycerol (TAG) components of OCO (23.93%), CCO (14.94%), LaCO (13.58%), OLaO (12.66%), and OOO (11.13%) were determined as the major TAG species in HMFS. The final optimal reaction conditions were carried out as follows: substrate molar ratio (oleic acid/CCSO), 5:1; enzyme concentration, 12.5% (w/w total reactants); reaction temperature, 60 °C; and reaction time, 28 h. The reusability of Lipozyme RM IM in the acidolysis reaction was also evaluated, and it was found that it could be reused up to 9 times without significant loss of activities. Urea inclusion method was used to separate and purify the synthetic product. As the ratio of HMFS/urea increased to 1:2, the acid value lowered to the minimum. In a scale-up experiment, the contents of TAG and total tocopherols in HMFS (modified CCSO) were 77.28% and 12.27 mg/100 g, respectively. All of the physicochemical indices of purified product were within food standards. Therefore, such a MCFA-enriched HMFS produced by using the acidolysis method might have potential application in the infant formula industry.