Flaxseed-derived peptides ameliorate hepatic cholesterol metabolism in Sprague-Dawley rats fed a high-cholesterol and high-fat diet.

BACKGROUND: Plant peptides have been reported to have cholesterol-lowering activities. Previous research has found that ≤1 kDa flaxseed peptide (FP5 ) reduces cholesterol absorption and synthesis in vitro. In this research, we investigated the cholesterol-lowering activity of FP5 in Sprague-Dawley (SD) rats fed a high-cholesterol and high-fat diet. In addition, amino acid sequences of FP5 were determined by high-performance liquid chromatography-electrospray ionization-Orbitrap mass spectrometry. RESULTS: FP5 supplement significantly decreased the serum and hepatic cholesterol levels and modulated the hepatic gene and protein expression of cholesterol metabolism-related enzymes or regulators (3-hydroxy-3-methylglutaryl coenzyme A reductase, Low-Density Lipoprotein Receptor (LDLR), Cholesterol 7 α-hydroxylase, Niemann-Pick C1-like 1, ATP-binding cassette transporters G5 and G8). Eleven peptides were identified from FP5 . These peptides were characterized as hydrophobic amino acids such as leucine (L), proline (P), glycine (G), isoleucine (I) and continuous sequences, including LP, LL, LG and II, with low molecular weights. CONCLUSION: FP5 has a certain cholesterol-lowering activity in SD rats fed a high-cholesterol and high-fat diet. The possible mechanism for ameliorating hepatic cholesterol metabolism of FP5 includes inhibiting hepatic cholesterol de novo synthesis, promoting the synthesis and excretion of bile acids, and inhibiting the reabsorption of bile acids during enterohepatic circulation. © 2022 Society of Chemical Industry.

Structural characterization of calcium-binding sunflower seed and peanut peptides and enhanced calcium transport by calcium complexes in Caco-2 cells.

BACKGROUND: Peptide-Ca complexes can promote Ca absorption. The present study aimed to determine the transport mechanism and structural characteristics of sunflower seed and peanut peptides with high Ca binding capacity with respect to developing third-generation Ca supplements and functional food ingredients. RESULTS: High Ca-binding fractions of 1-3 kDa sunflower seed peptide (SSP4 ) and ≥ 10 kDa peanut peptide (PP1 ) had higher amount of Ca transported than CaCl2 and two hydrolyzed proteins in Caco-2 cells. SSP4 and PP1 were separated by Ca ion metal chelate affinity chromatography, and high Ca-binding fractions were observed for SSP4 -P2 and PP1 -P2 . The amino acid sequences of SSP4 -P2 and PP1 -P2 were characterized by high-performance liquid chromatography-electrospray ionization-time of flight mass spectrometry. Seven and eight peptides were identified from SSP4 -P2 and PP1 -P2 , respectively. These peptides had molecular weights ranging from 1500 Da to 2500 Da and a large number of characteristic amino acid sequences, such as EEEQQQ, EQ-QQQ-QQ, QQ-QQQQQ, E-EEE, EE-EEQ, RR, Q-QQ-QQQ, EE-EQ-EE-Q, QQ-QQQQ, and Q-QQQQ, where 'E' is glutamic acid and 'Q' is glutamine. CONCLUSION: SSP4 and PP1 can promote Ca transport in Caco-2 cells without affecting cell permeability. The amino acid sequences of SSP4 -P2 and PP1 -P2 with high Ca-binding abilities contain characteristic sequences, such as continuous glutamic acid and glutamine, and have low molecular weights. © 2020 Society of Chemical Industry.

Ultrasound-assisted macroporous resin treatment improves the color and functional properties of sunflower meal protein.

Sunflower meal protein (SMP) has been considered as a high-quality source of plant protein. However, because the chlorogenic acid (CA) contained in sunflower seed meal was prone to oxidation reactions under traditional alkali extraction conditions, the extracted protein has a dark color and some poor functional properties. To this end, this study used ultrasound-assisted macroporous resin treatment to extract SMP. The improvement effects and potential mechanisms of ultrasonic-assisted macroporous resin treatment with different powers (100, 300, and 500 W) on the color and functional properties of SMP were studied. The results showed that compared with untreated sunflower meal protein (USMP), the lightness value (L*), solubility, emulsification, and gel elasticity were significantly enhanced when treated with 100 W and 300 W ultrasonic-assisted macroporous resin. However, when the ultrasonic power was increased to 500 W, the L* value, solubility, emulsification, and gel elasticity decreased instead, indicating that lower power (100 W and 300 W) ultrasonic-assisted macroporous resin treatment significantly improved the color and functional properties of SMP. Further research found that ultrasound-assisted macroporous resin treatment changed the secondary and tertiary structures of SMP, transformed ß-sheet into α-helix and ß-turn through rearrangement, and significantly improved surface hydrophobicity. It shows that ultrasonic-assisted macroporous resin treatment expands the SMP structure and exposes hydrophobic groups, thereby improving the color and functional properties of SMP. This study provides a potential strategy for extracting SMP with light color and good functional properties. It also provides a theoretical basis for the wide application of SMP in food processing.

Preparation of sunflower seed-derived umami protein hydrolysates and their synergistic effect with monosodium glutamate and disodium inosine-5'-monophosphate.

Sunflower seeds are rich in protein and can be an excellent raw material for the production of umami peptides. In this study, sunflower seed meal, which was defatted at a low temperature, was taken as the raw material, and proteins were separated, followed by hydrolyzation for 4 h by flavourzyme® to obtain hydrolysates with strong umami intensity. These hydrolysates were deamidated using glutaminase to increase the umami intensity. The highest umami value of 11.48 was recorded for hydrolysates deamidated for 6 h, and the umami intensity was determined. The umami hydrolysates mixed with 8.92 mmol IMP + 8.02 mmol MSG showed the highest umami value of 25.21. Different concentrations of ethanol were used for further separation of hydrolysates, and the highest umami value of 13.54 was observed for 20% ethanol fraction. The results of this study provide utilization method for sunflower seed meal protein and a theoretical basis for the preparation of umami peptides. PRACTICAL APPLICATION: A large number of sunflower seed meals after oil production are used as feed for livestock and poultry. Sunflower seed meal is rich in protein, and umami amino acid composition in sunflower seed meal is up to 25%-30%, which is potentially an excellent raw material for the production of umami peptides. The umami flavor and synergistic effect of obtained hydrolysates, with MSG and IMP, were analyzed in the present study. We intend to provide a novel way for utilization of protein from sunflower seed meal along with a theoretical basis for the preparation of umami peptides.

Flaxseed-derived peptide, IPPF, inhibits intestinal cholesterol absorption in Caco-2 cells and hepatic cholesterol synthesis in HepG2 cells.

Flaxseed peptides reduced serum cholesterol levels in Sprague-Dawley rats fed with a high-cholesterol diet. However, the mechanism of this action remains unclear. Flaxseed-hydrolyzed proteins were separated through ultrafiltration. The fifth fraction (FP5 , ≤ 1 kDa) had the highest cholesterol micelle solubility inhibition rate (CMSR) of 72.39% among the other fractions. Eleven peptides were identified from FP5 . Ile-Pro-Pro-Phe (IPPF), which had the highest CMSR of 93.47%, was selected for further analyses. IPPF substantially reduced the cholesterol transported content in Caco-2 cells and the total cholesterol content in HepG2 cells. Moreover, IPPF modulated the protein levels of NCP1L1 and ABCG5/8 (cholesterol transporters) in Caco-2 cells and reduced the mRNA levels of Srebp-2 and Hmgcr (cholesterol synthesis enzymes) in HepG2 cells. IPPF inhibits cholesterol intestinal absorption by modulating the cholesterol transporters expression and reduces hepatic cholesterol synthesis by inhibiting the SREBP2-regulated mevalonate pathway. IPPF is a new food-derived cholesterol-lowering nutritional supplement. PRACTICAL APPLICATIONS: We isolated active peptides with cholesterol-lowering properties from flaxseed protein, a by-product of industrial oil production, which greatly improved the economic and medicinal value of flaxseed protein. According to our research, IPPF can be used as a new food-derived type of cholesterol intestinal absorption inhibitor to reduce dietary cholesterol absorption and cholesterol synthesis inhibitor (same pharmacological mechanism as statins). IPPF provide a nutritional therapy component for hypercholesterolemia and prevent atherosclerosis. Our research provides theoretical basis for development and utilization of new nutritional supplements and plant proteins.

Purification and characterization of calcium-binding soybean protein hydrolysates by Ca2+/Fe3+ immobilized metal affinity chromatography (IMAC).

Soybean protein hydrolysates (SPHs) can bind calcium in order to form soluble peptide-calcium complexes. However, amino acid composition and structural characteristics of the calcium chelating SPHs are still unclear. This study separated SPHs with calcium and iron immobilized metal affinity chromatography (IMAC), and examined the effects of SPHs with different amino acid composition on calcium binding capacity. Three fractions (FFe-1, FFe-2 and FFe-3) isolated with IMAC-Fe(3+) were shown possessing increased Glu, Gln, Lys and Pro content from FFe-1 to FFe-3, and improved amount of bound calcium. Furthermore, the fractions adsorbed on IMAC-Ca(2+) (Fe(3+)) were separated and identified with reverse-phase (RP)-HPLC and MALDI-TOF MS/MS. The results showed that the sequence of peptides from FCa-2 and FFe-3 fractions was DEGEQPRPFPFP.

Identification and characteristics of iron-chelating peptides from soybean protein hydrolysates using IMAC-Fe3+.

The iron-chelating peptides from soybean protein hydrolysates (SPH) were investigated using immobilized metal affinity chromatography (IMAC). The results demonstrated that SPH could absorb on the IMAC-Fe(3+) column, while the capability of the binding iron was different in SPH (10-30 kDa), SPH (3-10 kDa), and SPH (1-3 kDa). The highest binding amount on the column occurred with SPH (10-30 kDa). With the IMAC method, the iron-chelating peptides were shown to be formed at pH lower than 5.5, and they were not affected by NaCL with the concentration between 0.1 mol/L and 1 mol/L, while the iron-chelating peptides could be partially disrupted by 0.02 mol/L Na(2)HPO(4) at pH 8.0. Furthermore, the iron-chelating peptides were identified with reversed phase (RP)-HPLC, SDS-PAGE, and MALDI-TOF MS/MS. The binding characteristics of the SPH on IMAC-Fe(3+) and the sequences of the iron-chelating peptides revealed that binding sites between SPH and iron might be the carboxyl groups of Glu and Asp residues.