Mouming Zhao

Purification and identification of antioxidant peptides from grass carp muscle hydrolysates by consecutive chromatography and electrospray ionization-mass spectrometry

Grass carp muscles were hydrolyzed with various proteases (papain, bovine pancreatin 6.0, bromelain, neutrase 1.5MG and alcalase 2.4L) to extract antioxidant peptides. The hydrolysates were assessed using methods of hydroxyl radical scavenging ability and lipid peroxidation inhibition activity. Hydrolysate prepared with alcalase 2.4L was found to have the highest antioxidant activity. It was purified using ultrafiltration and consecutive chromatographic methods including ion-exchange chromatography, multilayer coil high-speed counter-current chromatography, and gel filtration chromatography. The purified peptide, as a potent antioxidant, was identified as Pro-Ser-Lys-Tyr-Glu-Pro-Phe-Val (966.3Da) using RP-HPLC connected on-line to an electrospray ionization mass spectrometry. As well, it was found that basic peptides had greater capacity to scavenge hydroxyl radical than acidic or neutral peptides and that hydrophobic peptides contributed more to the antioxidant activities of hydrolysates than the hydrophilic peptides. In addition, the amino acid sequence of the peptide might play an important role on its antioxidant activity.

Identification of phenolics in the fruit of emblica (Phyllanthus emblica L.) and their antioxidant activities.

An activity-directed fractionation and purification process was used to identify the antioxidative components of emblica fruit. Dried fruit of emblica was extracted with methanol and then partitioned by ethyl ether, ethyl acetate, butanol and water. The ethyl acetate fraction showed the strongest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity among four fractions. The ethyl acetate fraction was then subjected to separation and purification using Sephadex LH-20 chromatography and reverse-phase high-performance liquid chromatography (HPLC). Six compounds were identified to be geraniin (1), quercetin 3-β-d-glucopyranoside (2), kaempferol 3-β-d-glucopyranoside (3), isocorilagin (4), quercetin (5), and kaempferol (6), respectively, by spectral methods, (1)H and (13)C nuclear magnetic resonance (NMR) spectroscopy, ultraviolet-visible (UV-Vis) spectrophotometry and mass spectroscopy (MS), and comparison with literatures. Compounds 2-4 and 6 were identified from emblica fruit for the first time. Furthermore, the antioxidant activities of purified compounds were screened for their antioxidative potential using lipid peroxidation and DPPH systems. All the purified compounds showed strong antioxidant and radical scavenging activities. Amongst, geraniin showed the highest antioxidant activity (4.7 and 65.7μM of IC50 values for DPPH and lipid peroxidation assay, respectively) than other purified compounds.

Effects of ultrasound pretreatment on the enzymatic hydrolysis of soy protein isolates and on the emulsifying properties of hydrolysates.

Soy protein isolate (SPI) was modified by ultrasound pretreatment (200 W, 400 W, 600 W) and controlled papain hydrolysis, and the emulsifying properties of SPIH (SPI hydrolysates) and USPIH (ultrasound pretreated SPIH) were investigated. Analysis of mean droplet sizes and creaming indices of emulsions formed by SPIH and USPIH showed that some USPIH had markedly improved emulsifying capability and emulsion stabilization against creaming during quiescent storage. Compared with control SPI and SPIH-0.58% degree of hydrolysis (DH), USPIH-400W-1.25% (USPIH pretreated under 400W sonication and hydrolyzed to 1.25% DH) was capable of forming a stable fine emulsion (d43=1.79 μm) at a lower concentration (3.0% w/v). A variety of physicochemical and interfacial properties of USPIH-400W products have been investigated in relation to DH and emulsifying properties. SDS-PAGE showed that ultrasound pretreatment could significantly improve the accessibility of some subunits (α-7S and A-11S) in soy proteins to papain hydrolysis, resulting in changes in DH, protein solubility (PS), surface hydrophobicity (H0), and secondary structure for USPIH-400W. Compared with control SPI and SPIH-0.58%, USPIH-400W-1.25% had a higher protein adsorption fraction (Fads) and a lower saturation surface load (Γsat), which is mainly due to its higher PS and random coil content, and may explain its markedly improved emulsifying capability. This study demonstrated that combined ultrasound pretreatment and controlled enzymatic hydrolysis could be an effective method for the functionality modification of globular proteins.

Enhanced antioxidant and antityrosinase activities of longan fruit pericarp by ultra-high-pressure-assisted extraction.

The health benefits of fruits acting against chronic diseases are ascribed to their antioxidant activities which are mainly responsible due to the presence of phenolic compounds. The use of ultra-high-pressure-assisted extraction (UHPE) has shown great advantages for the extraction of these phenolic compounds from longan fruit pericarp (LFP). Studies were carried out to investigate the effects of UHPE at pressures of 200, 300, 400 and 500 MPa on total phenolic contents, extraction yield, antioxidant and antityrosinase activities from LFP. The antioxidant activities of these extracts were analyzed, using various antioxidant models like 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, total antioxidant capacity and superoxide anion radical scavenging activity. Extract from ultra-high-pressure-assisted extraction at 500MPa (UHPE-500) showed the highest antioxidant activities of all the tested models. In addition, it also showed moderate tyrosinase inhibitory activity. Three phenolic acids, namely gallic acid, ellagic acid, and corilagin were identified and quantified by HPLC. Corilagin content was the highest compared to other phenolic acids identified. UHPE-500 obtained the higher phenolic acid contents compared to other high pressure processing and conventional extractions (CE). Compared with CE, UHPE-500 exhibited good extraction effectiveness in terms of higher extraction yields with high phenolic contents and also with higher antioxidant and antityrosinase activities.

Structural characterisation of polysaccharides from Tricholoma matsutake and their antioxidant and antitumour activities

In this study, polysaccharides from Tricholoma matsutake (TM-P) were purified using a DEAE-Sepharose fast flow column and three polysaccharide fractions (TM-P1, TM-P2 and TM-P3) were obtained. The chemical composition and structural characteristics of TM-P2 were quite different from those of TM-P1 and TM-P3. TM-P2 consisted of glucose, galactose and mannose with a molar ratio of 5.9:1.1:1.0. The glycosidic linkages were mainly composed of 1,6- and 1-linked glucose. Furthermore, TM-P2 showed the strongest in vitro antioxidant and antitumour activities. The oxygen radical absorbance capacity (ORAC) of TM-P2 was 2100.44 μmol Trolox/g. The antiproliferative activities of TM-P2 (4.0mg/ml) on the growth of HepG2 and A549 cells were 67.98% and 59.04%, respectively.

Application of targeted drug delivery system in Chinese medicine

Targeted drug delivery system of traditional Chinese medicine (TCM) refers to those using different carriers to make the effective parts or monomer extracted from TCM or natural medicine into agents which can directly concentrate on the target site. This system is an ideal delivery approach and has became a hot spot in the field of TCM pharmaceutical research since it can improve the pharmacological effects and reduce the adverse reactions. This paper reviews literatures on TCM targeted agents which were published in the past 10 years. In accordance with the different carriers, four types of agents, liposome, nanoparticle, microsphere, and emulsion are analyzed. Liposomes were studied most profoundly and a variety of new types of liposomes was developed on the basis of the traditional liposomes. Using natural or synthetic polymer materials to carry drugs, nanoparticles and microspheres can promote the drug through the blood-brain barrier and enhance its bioavailability. Emulsion has lymphatic affinity and the drug is coated in the internal phase, which can protect the drugs from hydrolysis. All these delivery agents are proved to be effective ways to improve the clinical efficacy of drugs, and each is discussed in detail with examples. At present, TCM targeted agents are still in the exploratory stage and many problems need to be solved. Especially, it is a huge challenge to research the targeted delivery systems for the effective parts of Chinese medicines and compound prescriptions, and the paper gives a particular discussion on this point. In the future, more attention should be paid to the research on the particle agents of TCM effective parts, and the development of new carrier materials in order to enhance the overall quality of TCM targeted agents.

Effect of ultrasonic treatment on the graft reaction between soy protein isolate and gum acacia and on the physicochemical properties of conjugates.

Soy protein isolate (SPI) was grafted with gum acacia (GA) in this work. In comparison with classical heating at high water activity, ultrasound could accelerate the graft reaction between SPI and GA. A degree of graft (DG) of 34 was obtained by ultrasonic treatment for 60 min, whereas 48 h was required with classical heating. Ultrasonic treatment improved the concentration of available free amino groups of SPI. The grafted SPI showed significantly (p < 0.05) higher levels of emulsifying activity index, emulsifying stability index, and surface hydrophobicity than native SPI. The droplet size (D[3,2] and D[4,3]) of SPI emulsion decreased from 8.3 to 2.3 microm and from 25.7 to 7.1 microm through ultrasound-assisted grafting, respectively. Moreover, SPI-GA conjugates gave emulsions exhibiting more stability against creaming than those prepared with only SPI during ambient temperature storage (20 days). Decreases of lysine and arginine contents during the graft reaction indicated that these two amino acid residues attended the covalent linkage between SPI and GA. The results of secondary structure suggested that grafted SPI had decreased alpha-helix and beta-sheet levels and increased unordered coils level.

Isolation and characterization of an oxygen radical absorbance activity peptide from defatted peanut meal hydrolysate and its antioxidant properties.

Defatted peanut meal hydrolysate (DPMH) was purified using ultrafiltration, gel filtration chromatography, and high-performance liquid chromatography. A tripeptide with strong oxygen radical absorbance capacity (ORAC) was isolated and identified as Tyr-Gly-Ser by ESI-MS/MS. It was then synthesized to measure its antioxidant properties in different systems. The ORAC value of Tyr-Gly-Ser was 3-fold higher than that of glutathione (GSH), and it displayed a stronger protective effect on linoleic acid peroxidation and H(2)O(2)-induced oxidative injury in rat pheochromocytoma line PC12 cells than GSH (p < 0.05). However, Tyr-Gly-Ser showed negligible DPPH radical scavenging activity, reducing power, and no metal chelating ability. The results suggested that Tyr-Gly-Ser displayed antioxidant activity via the hydrogen atom transfer mechanism, and the Tyr at the N-terminal was the hydrogen donor. The ORAC assay was recommended as a reliable and effective method to measure the antioxidant activity in the course of antioxidant peptide isolation.

Effect of maillard reaction products derived from the hydrolysate of mechanically deboned chicken residue on the antioxidant, textural and sensory properties of Cantonese sausages.

Protein hydrolysates as precursors of Maillard reaction were obtained via enzymatic hydrolysis of mechanically deboned chicken residue (MDCR). The Maillard reaction products (MRPs) were prepared at 90 (M1), 100 (M2), 110 (M3) and 120 degrees C (M4), respectively. MRPs possessed a strong reducing power and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. According to the evolution of total free fatty acids and peroxide value of Cantonese sausage with MRPs during storage, M1 and M3 had a potent antioxidant activity (P<0.05) due to their antioxidant abilities and inhibitory action against lipolytic enzymes. Cantonese sausages treated with M1 and M2 showed good textural and sensory properties. However, M3 and M4 had a negative (P<0.05) effect on the flavour and texture of Cantonese sausages compared to control. The results suggested that M1 was very potential to be used to improve their antioxidant, textural and sensory quality.

Identification of a novel phenolic compound in litchi (Litchi chinensis Sonn.) pericarp and bioactivity evaluation

Litchi (Litchi chinensis Sonn.) is a delicious fruit widely accepted by consumers all over the world. In this work, phytochemical investigation of litchi pericarp methanol extracts led to the isolation of a novel phenolic, 2-(2-hydroxyl-5-(methoxycarbonyl) phenoxy) benzoic acid, together with kaempferol, isolariciresinol, stigmasterol, butylated hydroxytoluene, 3,4-dihydroxyl benzoate, methyl shikimate and ethyl shikimate. Most were found in litchi pericarp for the first time. Their structures were mainly elucidated by NMR and MS evidences. Antioxidant activities of the eight compounds were determined by a DPPH radical scavenging assay and the results showed that 2-(2-hydroxy-5-(methoxycarbonyl) phenoxy)benzoic acid, kaempferol, isolariciresinol, butylated hydroxytoluene and 3,4-dihydroxy benzoate exhibited good antioxidant activities. An interesting finding was that butylated hydroxytoluene was detected as a natural antioxidant in this work, which was usually taken as a synthesized antioxidant. Furthermore, the novel compound exhibited no inhibitory effects against tyrosinase and α-glucosidase activities.