Major phenolics in apple and their contribution to the total antioxidant capacity

The contribution of each phytochemical to the total antioxidant capacity of apples was determined. Major phenolic phytochemicals of six apple cultivars were identified and quantified, and their contributions to total antioxidant activity of apples were determined using a 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging assay and expressed as vitamin C equivalent antioxidant capacity (VCEAC). Average concentrations of major phenolics and vitamin C in six apple cultivars were as follows (mg/100 g of fresh weight of apples): quercetin glycosides, 13.20; procyanidin B(2), 9.35; chlorogenic acid, 9.02; epicatechin, 8.65; phloretin glycosides, 5.59; vitamin C, 12.80. A highly linear relationship (r (2) > 0.97) was attained between concentrations and total antioxidant capacity of phenolics and vitamin C. Relative VCEAC values of these compounds were in the order quercetin (3.06) > epicatechin (2.67) > procyanidin B(2) (2.36) > phloretin (1.63) > vitamin C (1.00) > chlorogenic acid (0.97). Therefore, the estimated contribution of major phenolics and vitamin C to the total antioxidant capacity of 100 g of fresh apples is as follows: quercetin (40.39 VCEAC) > epicatechin (23.10) > procyanidin B(2) (22.07) > vitamin C (12.80) > phloretin (9.11) > chlorogenic acid (8.75). These results indicate that flavonoids such as quercetin, epicatechin, and procyanidin B(2) rather than vitamin C contribute significantly to the total antioxidant activity of apples.     

Contribution of individual polyphenolics to total antioxidant capacity of plums

To study the effect of polyphenolics on antioxidant capacities of plums, the amounts of total phenolics, total flavonoids and individual phenolic compounds, and vitamin C equivalent antioxidant capacity (VCEAC) of eleven plum cultivars was determined. There was a good linear relationship between the amount of total phenolics and total antioxidant capacity (r2 = 0.9887). The amount of total flavonoids and total antioxidant capacity also showed a good correlation (r2 = 0.9653). Although the summation of individual antioxidant capacity was lower than the total antioxidant capacity of plum samples, there was a positive correlation (r2 = 0.9299) of total antioxidant capacity of plum samples with the sum of the VCEACs calculated from individual phenolics. Chlorogenic acids and glycosides of cyanidin, peonidin, and quercetin were major phenolics among eleven plum cultivars. The antioxidant capacity of chlorogenic acids and anthocyanins showed higher correlation (r2) of 0.7751 and 0.6616 to total VCEAC, respectively, than that of quercetin glycosides (r2 = 0.0279). Chlorogenic acids were a major source of antioxidant activity in plums, and the consumption of one serving (100 g) of plums can provide antioxidants equivalent to 144.4-889.6 mg of vitamin C.     

Cloning, expression, and characterization of two beta-glucosidases from isoflavone glycoside-hydrolyzing Bacillus subtilis natto

On the basis of the genomic sequence of Bacillus subtilis 168, two beta-glucosidase genes (bglH and yckE) from B. subtilis natto, which has been reported to have high isoflavone glucoside-hydrolyzing activity, were cloned and overexpressed in E. coli M15. The temperature for the optimal p-nitrophenyl-beta-D-glucoside hydrolyzing activity of both enzymes was between 37 and 45 degrees C, but BglH had a higher thermal stability than YckE. Both showed high activity at pH 6.0, but YckE was stable over a wider pH range than BglH. Recombinant BglH was inhibited 73%, 63%, and 43% by 1.0 mM Cd(2+), Fe(2+), or Cu(2+), respectively, while other divalent metal ions resulted in 0-23% inhibition, whereas YckE was inhibited by less than 20% by any of the divalent metal ions we tested. Among the substrate we used, BglH showed the highest affinity for genistin and YckE showed the highest affinity for p-nitrophenyl-beta-D-fructopyranoside. Both BglH and YckE hydrolyzed genistin and daidzin into their isoflavone aglycones, genistein and daidzein, but BglH was more efficient than YckE in isoflavone glucoside hydrolysis (20-fold higher kcat). Our results suggest that recombinant BglH may be applicable in the process of isoflavones deglycosylation.     

Characterization of volatile aroma compounds in cooked black rice

Black rice ( Oryza sativa L.), an aromatic specialty rice popular in Asia, has a unique flavor, the volatile chemistry of which has not been reported. The objectives of this research were to study volatile profiles of cooked black rice and to characterize the odor-active compounds. Thirty-five volatile compounds were identified by gas chromatography-mass spectrometry using a dynamic headspace system with Tenax trapping. Aldehydes and aromatics were quantitatively in the greatest abundance, accounting for 80.1% of total relative concentration of volatiles. The concentration of 2-acetyl-1-pyrroline (2-AP) was high, exceeded only by hexanal, nonanal, and 2-pentylfuran. A total of 25 odor-active compounds, determined by gas chromatography-olfactometry, were applied to principal component analysis, demonstrating significant differences between a black and a traditional white rice cultivar in terms of aroma and explaining 93.0% of the total variation. 2-AP, guaiacol, indole, and p-xylene largely influenced the difference between the aroma in cooked black and white rice. 2-AP and guaiacol were major contributors to the unique character of black rice based on odor thresholds, relative concentrations, and olfactometry.     

Latent and active polyphenol oxidase (PPO) in red clover (Trifolium pratense) and use of a low PPO mutant to study the role of PPO in proteolysis reduction

Polyphenol oxidase (PPO) activity in leaf extracts of wild type (WT) red clover and a mutant line expressing greatly reduced levels of PPO (LP red clover) has been characterized. Both latent and active forms of PPO were present, with the latent being the predominant form. PPO enzyme and substrate (phaselic acid) levels fluctuated over a growing season and were not correlated. Protease activation of latent PPO was demonstrated; however, the rate was too low to have an immediate effect following extraction. A novel, more rapid PPO activation mechanism by the enzyme's own substrate was identified. Rates of protein breakdown and amino acid release were significantly higher in LP red clover extracts compared with WT extracts, with 20 versus 6% breakdown of total protein and 1.9 versus 0.4 mg/g FW of free amino acids released over 24 h, respectively. Inclusion of ascorbic acid increased the extent of protein breakdown. Free phenol content decreased during a 24 h incubation of WT red clover extracts, whereas protein-bound phenol increased and high molecular weight protein species were formed. Inhibition of proteolysis occurred during wilting and ensilage of WT compared with LP forage (1.9 vs 5 and 17 vs 21 g/kg of DM free amino acids for 24 h wilted forage and 90 day silage, respectively). This study shows that whereas constitutive red clover PPO occurs predominantly in the latent form, this fraction can contribute to reducing protein breakdown in crude extracts and during ensilage.     

Antioxidant effects of phlorotannins isolated from Ishige okamurae in free radical mediated oxidative systems

Three phlorotannins, including phloroglucinol, diphlorethohydroxycarmalol, and 6,6'-bieckol, were isolated from Ishige okamurae by column chromatography. The structures of the phlorotannins were determined on the basis of spectroscopic analysis, including NMR and mass spectrometry (MS) techniques. Antioxidant effects of phlorotannins were measured by direct free radical scavenging activities using the electron spin resonance spectrometry (ESR) technique and cellular systems in vitro. The results indicated that diphlorethohydroxycarmalol and 6,6'-bieckol showed potential radical scavenging activities against the 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, alkyl, and superoxide radicals. Moreover, no cytotoxicities of the phlorotannins on human fetal lung fibroblasts cell line (MRC-5), mouse macrophages cell line (RAW264.7), and human leukemic cell line (HL-60) were observed. In addition, diphlorethohydroxycarmalol and 6,6'-bieckol significantly reduced the intracellular reactive oxygen species level assessed by 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay in RAW264.7 cells, and myeloperoxide (MPO) activity in HL-60 cells and radical-mediated oxidation of cell membrane proteins in RAW264.7 cells were dose-dependently inhibited in the presence of diphlorethohydroxycarmalol and 6,6'-bieckol. In conclusion, these results suggested that phlorotannins could be used as novel functional foodstuffs or antioxidants in the cosmetic and drug industries.     

Nematicidal activity of plant essential oils and components from coriander (Coriandrum sativum), Oriental sweetgum (Liquidambar orientalis), and valerian (Valeriana wallichii) essential oils against pine wood nematode (Bursaphelenchus xylophilus)

Commercial essential oils from 28 plant species were tested for their nematicidal activities against the pine wood nematode, Bursaphelenchus xylophilus. Good nematicidal activity against B. xylophilus was achieved with essential oils of coriander (Coriandrum sativum), Oriental sweetgum (Liquidambar orientalis), and valerian (Valeriana wallichii). Analysis by gas chromatography-mass spectrometry led to the identification of 26, 11, and 4 major compounds from coriander (Coriandrum sativum), Oriental sweetgum (Liquidambar orientalis), and valerian (Valeriana wallichii) oils, respectively. Compounds from each plant essential oil were tested individually for their nematicidal activities against the pine wood nematode. Among the compounds, benzaldehyde, trans-cinnamyl alcohol, cis-asarone, octanal, nonanal, decanal, trans-2-decenal, undecanal, dodecanal, decanol, and trans-2-decen-1-ol showed strong nematicidal activity. The essential oils described herein merit further study as potential nematicides against the pine wood nematode.     

Tastes and structures of bitter peptide, asparagine-alanine-leucine-proline-glutamate, and its synthetic analogues

Asn-Ala-Leu-Pro-Glu (NALPE) is a strong bitter peptide with a minimum response threshold (MRT) of 0.074 mM. To elucidate the relationship of spatial structure and bitterness on peptides, NALPE and its analogues, NALPW, NALPS, NALPL, NALPP, NALPD, and NALPR, were synthesized and sensorially evaluated. Structural analysis using computer simulation for each peptide revealed that the presence of a polar group and hydrophobic bitter amino acids, the composition of hydrophobic regions, the spatial orientation of the polar group and hydrophobic regions, and the proximity between polar groups and hydrophobic regions faced within the same plane space may be the major determinants for the taste type and intensity of peptide bitterness.     

Release kinetics of tocopherol and quercetin from binary antioxidant controlled-release packaging films

This paper investigated the feasibility of manipulating packaging polymers with various degrees of hydrophobicity to release two antioxidants, tocopherol and quercetin, at rates suitable for long-term inhibition of lipid oxidation in food. For example, one antioxidant can be released at a fast rate to provide short-term/intermediate protection, whereas the other antioxidant can be released at a slower rate to provide intermediate/long-term protection of lipid oxidation. Controlled-release packaging films containing tocopherol and quercetin were produced using ethylene vinyl alcohol (EVOH), ethylene vinyl acetate (EVA), low-density polyethylene (LDPE), and polypropylene (PP) polymers; the release of these antioxidants to 95% ethanol (a fatty food simulant) was measured using UV-vis spectrophotometry, and Fickian diffusion models with appropriate initial and boundary conditions were used to fit the data. For films containing only quercetin, the results show that the release of quercetin was much faster but lasted for a much shorter time for hydrophilic polymers (EVOH and EVA) than for hydrophobic polymers (LDPE and PP). For binary antioxidant films containing tocopherol and quercetin, the results show that tocopherol released more rapidly but for a shorter period of time than quercetin in LDPE and EVOH films, and the difference is more pronounced for LDPE films than EVOH films. The results also show the presence of tocopherol can accelerate the release of quercetin. Although none of the films produced is acceptable for long-term lipid oxidation inhibition, the study provides encouraging results suggesting that acceptable films may be produced in the future using polymer blend films.