Yuanying Ni

Comparison of membrane-bound and soluble polyphenol oxidase in Fuji apple (Malus domestica Borkh. cv. Red Fuji)

This study compared membrane-bound with soluble polyphenol oxidase (mPPO and sPPO, respectively) from Fuji apple. Purified mPPO and partially purified sPPO were used. mPPO was purified by temperature-induced phase partitioning and ion exchange chromatography. The specific activity of mPPO was 34.12× higher than that of sPPO. mPPO was more stable than sPPO at pH 5.0-8.5. Although mPPO was more easily inactivated at 25-55 °C, it is still more active than sPPO in this temperature range. The optimum substrate of mPPO was 4-methyl catechol, followed by catechol. L-cysteine had the highest inhibitory effects on mPPO followed by ascorbic acid and glutathione. Surprisingly, EDTA increased mPPO activity. The results revealed that purified mPPO is a dimer with a molecular weight of approximately 67 kDa.

Separation and preparation of 6-gingerol from molecular distillation residue of Yunnan ginger rhizomes by high-speed counter-current chromatography and the antioxidant activity of ginger oils in vitro

Molecular distillation residue (MD-R) from ginger had the most total phenol content of 247.6mg gallic acid equivalents per gram (GAE/g) among the ginger oils. High-speed counter-current chromatography (HSCCC) technique in semi-preparative scale was successfully performed in separation and purification of 6-gingerol from MD-R by using a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (10:2:5:7, v/v/v/v). The target compound was isolated, collected, purified by HSCCC in the head-tail mode, and then analyzed by HPLC. A total of 90.38±0.53mg 6-gingerol was obtained from 600mg MD-R, with purity of 99.6%. In addition, the structural identification of 6-gingerol was performed by EI/MS, (1)H NMR and (13)C NMR. Moreover, the orders of antioxidant activity were vitamin E (VE)>supercritical fluid extraction oleoresin (SFE-O)=MD-R=6-gingerol>molecular distillation essential oil (MD-EO) and butylated hydroxytoluene (BHT)=VE>6-gingerol>MD-R=SFE-O>MD-EO, respectively in 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging and β-Carotene bleaching.

Purification and structural analysis of membrane-bound polyphenol oxidase from Fuji apple.

Membrane-bound polyphenol oxidase (mPPO) in Fuji apple (Malus domestica Borkh. cv. Red Fuji) was purified and analyzed with a nanoelectrospray ionization mass spectrometer. The three-dimensional model and binding site of mPPO to 4-methyl catechol were also studied using molecular docking. mPPO was purified 54.41-fold using temperature-induced phase partitioning technique and ion exchange chromatography. mPPO had a molecular weight of 67.3kDa. Even though a significant level of homology was observed between mPPO and the soluble polyphenol oxidase in the copper binding sequence, there was another region, rich in histidine residues, which differed in 13 amino acids. The three-dimensional structure of mPPO consisted of six α-helices, two short β-strands, and ten random coils. The putative substrate-binding pocket contained six polar or charged amino acids, His191, His221, Trp224, Trp228, Phe227, and Val190. Trp224 and Trp228 formed hydrogen bonds with 4-methyl-catechol.

Characterisation of seed oils from different grape cultivars grown in China

To explore the potential of the large amount of grape pomace in wineries of China, oils of three Eurasianxa0grape cultivars (Chardonnay, Merlot and Carbernet Sauvignon) and two Chinese traditional grape cultivars (Vitis amurensis and Vitis davidii), were characterised. The results showed seed oil properties differ for various grape varities. Grape seed oils were demonstrated to be good sources of polyunsaturated fatty acid (PUFA) (63.88–77.12xa0%), sterols (227.99–338.83xa0mg/100xa0g oil) and tocotrienols (320.08–679.24xa0mg/kg oil). Seed oil of V. amurensis exhibited the highest values of polyunsaturated fatty acid, total tocotrienols, total tocols and DPPH· scavenging capacity. Seed oil of Carbernet Sauvignon had the highest contents of squalene, total sterols, total tocopherols and total phenolics. Principal component analysis five grape cultivars differentiated on the basis of bioactive components content and antioxidant properties.

Carotenoids and Carotenoid Esters of Red and Yellow Physalis (Physalis alkekengi L. and P. pubescens L.) Fruits and Calyces

Carotenoid profiles of fruits and calyces of red (Physalis alkekengi L.) and yellow (P. pubescens L.) Physalis were characterized by HPLC-DAD-APCI-MSn. Altogether 69 carotenoids were detected in red Physalis, thereof, 45 were identified. In yellow Physalis, 40 carotenoids were detected and 33 were identified. Zeaxanthin esters with various fatty acids were found to be the most abundant carotenoids in red Physalis, accounting for 51-63% of total carotenoids, followed by β-cryptoxanthin esters (16-24%). In yellow Physalis, mainly free carotenoids such as lutein and β-carotene were found. Total carotenoid contents ranged between 19.8 and 21.6 mg/100 g fresh red Physalis fruits and 1.28-1.38 mg/100 g fresh yellow Physalis fruits, demonstrating that Physalis fruits are rich sources of dietary carotenoids. Yellow Physalis calyces contained only 153-306 μg carotenoids/g dry weight, while those of red Physalis contained substantially higher amounts (14.6-17.6 mg/g dry weight), thus possibly exhibiting great potential as a natural source for commercial zeaxanthin extraction.

Optimization of ultrasound-assisted extraction of okra (Abelmoschus esculentus (L.) Moench) polysaccharides based on response surface methodology and antioxidant activity

This study determined the optimal conditions for ultrasound-assisted extraction of a water-soluble polysaccharide, Raw Okra Polysaccharide, from the fruit of okra using response surface methodology. The optimal extraction temperature, extraction time and ultrasonic power were 59°C, 30min and 522W, respectively, giving a yield of 10.35±0.11%. ROP was further isolated, lyophilized and purified using a DEAE-Sepharose Fast Flow column and Sepharose CL-6B column, revealing three elution peaks subsequently designated ROP -1, -2, and -3, respectively. Of these, ROP-2 showed the highest yield, and was therefore selected for physicochemical analysis and evaluation of antioxidant activity. Gas chromatography, fourier transform infrared spectroscopy, and high-performance liquid chromatography were used to characterize the primary structural features and molecular weight, revealing that ROP-2 is composed of glucose, mannose, galactose, arabinose, xylose, fructose, and rhamnose (molar percentages: 28.8, 12.5, 13.1, 15.9, 9.2, 13.7, and 6.8%, respectively) and has an average molecular weight of 1.92×105Da. A superoxide radical scavenging assay and DPPH radical scavenging assay further revealed the significant in vitro antioxidant activity of ROP-2. These findings present an effective technique for extraction of the natural antioxidant ROP-2, warranting further analysis of its potential application in the food industry.

Sulfated modification and anticoagulant activity of pumpkin (Cucurbita pepo, Lady Godiva) polysaccharide

Sulfated modification of pumpkin polysaccharide using CAS with pyridines as catalysts under different conditions was conducted to obtain different degrees of sulfation on a laboratory scale. Anticoagulant activities of pumpkin polysaccharide and its sulfated derivatives were also investigated employing various established in vitro systems. Results showed that addition of high ratio of CAS/pyridine under constant conditions could increase the degree of substitution. Sulfate substitution was further confirmed by the FT-IR and 13C NMR analysis. The df values between 2.11-2.73 indicated the relatively expanded conformation of the sulfated derivatives. The sulfated polysaccharides showed higher anticoagulant activities through activated partial thrombosis time (aPTT), thrombin time (TT), prothrombin time (PT) and anti-Xa activity assay, which revealed that better anticoagulant activities could be obtained when DS remained higher and Mw maintained in a moderate range.

Comparison of biochemical properties and thermal inactivation of membrane-bound polyphenol oxidase from three apple cultivars (Malus domestica Borkh)

A comparative study of the properties of membrane‐bound polyphenol oxidase (mPPO) from three apple cultivars, namely Red Fuji (FJ), Granny Smith (GS) and Golden Delicious (GD), was carried out for the first time. Data indicate that mPPOs from three cultivars exhibit significantly different properties. GS mPPO had the strongest affinity to catechol, but FJ mPPO had the highest maximum velocity. Red Fuji (FJ) mPPO had the significantly higher activity than those of GD and GS mPPOs. Red Fuji (FJ) mPPO had the highest activity at pH 8.00, while GD and GS mPPOs at 4.50 and 7.50–8.00, respectively. Red Fuji (FJ) mPPO was more stable than GD and GS mPPOs over the pH range of 5.0–8.5. The optimal temperature for GS mPPO was within 70–75xa0°C, which is higher than those for mPPOs from FJ and GD. Thermal inactivation of the three mPPOs followed a first‐order kinetic model with different inactivation kinetic parameters.

Screening of critical factors influencing the efficient hydrolysis of zeaxanthin dipalmitate in an adapted in vitro- digestion model

As hydrolysis of carotenoid esters is believed to be highly efficient in vivo, their insufficient hydrolysis in in vitro-digestion models, particularly, regarding zeaxanthin diesters, is a current issue. Therefore, in this study, several factors related to the enzymatic hydrolysis were investigated in an adapted version of the standardized INFOGEST in vitro-digestion model, using zeaxanthin dipalmitate (ZDP) as a substrate. The results showed that pancreatic lipase was able to hydrolyze ZDP, whereas carboxyl ester lipase (CEL) substantially contributed to ZDP cleavage. Replacement of commonly used porcine with bovine bile extracts and the substitution of coffee creamer for soybean oil at identical fat contents both significantly improved hydrolysis efficiency and bioaccessibility of total zeaxanthin to better mimic in vivo conditions. Thus, bile and lipids selection for in vitro digestion of carotenoid esters was crucial. The combined use of coffee creamer, pancreatin, CEL, and bovine bile led to the highest hydrolysis efficiency of 29.5%.