Ryoji Takata

Environmental stress enhances biosynthesis of flavor precursors, S-3-(hexan-1-ol)-glutathione and S-3-(hexan-1-ol)-L-cysteine, in grapevine through glutathione S-transferase activation

The biosynthesis of S-(3-hexan-1-ol)-glutathione (3MH-S-glut) and S-(3-hexan-l-ol)-L-cysteine (3MH-S-cys), which act as flavour precursors in wines, in Vitis vinifera grapes exposed to various environmental stress conditions is reported here. Ultraviolet (UV-C) irradiation, water deficit, and biological stimulation up-regulated 3MH-S-glut and 3MH-S-cys biosynthesis in grape leaves. 3MH-S-glut and 3MH-S-cys contents in grape berries were increased by cold shock, heat shock, UV-C irradiation, and biological stimulation. The results suggest that environmental stress enhances the biosynthesis of both flavour precursors in grapevine. The transcription of VvGST1, VvGST3, VvGST4, and GGT in grapevine exposed to the stress conditions was increased markedly compared with that in control grapevine. Also, UV irradiation increased GST (glutathione S-transferase) and GGT (γ-glutamyl transferase) enzyme activities in grape berries. Recombinant VvGST3 and VvGST4, but not VvGST1, mediated the synthesis of 3MH-S-glut from reduced glutathione and trans-2-hexenal in vitro. The enzymatic mediation of flavour precursor production is a novel function of plant GSTs and may result in the detoxification of damaged grape cells under stress conditions.

Immunostimulatory effects of a polysaccharide-rich substance with antitumor activity isolated from black currant (Ribes nigrum L.).

The fruit juice of black currant was found to contain a polysaccharide-rich substance, which was designated cassis polysaccharide (CAPS), with macrophage-stimulating activity. Especially, its interleukin (IL)-1β-inducing activity was remarkably high, compared with other fruit juice preparations. CAPS was found to consist of rhamnose, mannose, arabinose, galactose, xylose, and glucose in a molar ratio of 11.3:0.9:54.1:29.8:2.0:1.9. CAPS turned out to be partitioned into a soluble component (CAPS-l.m.) and a precipitable component (CAPS-h.m.) with mean MWs of 80,000 and 600,000 respectively in 45% (v/v) ethanol solution. At least in vitro, CAPS-l.m. rather than CAPS-h.m. appeared to play an important role in macrophage activation. Oral administration of black currant juice and CAPS to Ehrlich carcinoma-bearing mice retarded the growth of the solid tumor by 45% and 51% respectively. CAPS administration had a stimulatory effect on the release of IL-2, IL-10, interferon-γ, and IL-4 from splenocytes in comparison with PBS treatment in tumor-bearing mice. The IL-4 level was, however, still lower than that exhibited by a group of normal mice. CAPS showed a certain cytotoxicity directly against tumor cells.

Cytochrome P450 CYP71BE5 in grapevine (Vitis vinifera) catalyzes the formation of the spicy aroma compound (−)-rotundone

Highlight CYP71BE5 from grapevine was identified as a sesquiterpene oxidase capable of transforming α-guaiene to (−)-rotundone, responsible for the characteristic spicy aroma in wines.

Iron is an essential cause of fishy aftertaste formation in wine and seafood pairing.

Fishy aftertaste is sometimes perceived in wine with fish and seafood pairing. However, what component of wine clashes with seafood or what compound contributes to the unpleasant fishy aftertaste in the mouth remains an open problem. First, intensities of unpleasant fishy aftertaste of wine and dried scallop pairings were rated by sensory analysis. Second, components of the wines were analyzed. Strong positive correlations were found between the intensity of fishy aftertaste and the concentration of both total iron and ferrous ion. Moreover, the intensity of fishy aftertaste was increased by the addition of ferrous ion in model wine and suppressed by the chelation of ferrous ion in red wine. Third, potent volatile compounds of fishy aftertaste, such as hexanal, heptanal, 1-octen-3-one, (E,Z)-2,4-heptadienal, nonanal, and decanal, were determined by gas chromatography-olfactometry and gas chromatography-mass spectrometry in dried scallop soaked in red wine. The formations of these compounds depended on the dose of ferrous ion in the model wine. These results suggest that ferrous ion is a key compound of the formation of fishy aftertaste in wine and seafood pairing within the concentration range commonly found in wine.

Improvement of the Antitumor Activity of Black Currant Polysaccharide by an Enzymatic Treatment

A polysaccharide-rich substance isolated from black currant, named cassis polysaccharide (CAPS), was partially digested with β-galactosidase from Aspergillus oryzae and its immunostimulatory activity was investigated. The in vitro cytokine-inducing effect of CAPS on RAW264 cells was gradually decreased along with lowering of the average MW of CAPS. In vivo, partially digested CAPS with a mean MW of approximately 20,000 showed the most potent antitumor activity against Ehrlich carcinoma in mice.

Systems pharmacology of adiposity reveals inhibition of EP300 as a common therapeutic mechanism of caloric restriction and resveratrol for obesity

Both caloric restriction (CR) and resveratrol (RSV) have beneficial effects on obesity. However, the biochemical pathways that mediate these beneficial effects might be complex and interconnected and have not been fully elucidated. To reveal the common therapeutic mechanism of CR and RSV, we performed a comparative transcriptome analysis of adipose tissues from diet-induced obese (DIO) zebrafish and obese humans. We identified nine genes in DIO zebrafish and seven genes in obese humans whose expressions were regulated by CR and RSV. Although the gene lists did not overlap except for one gene, the gene ontologies enriched in the gene lists were highly overlapped, and included genes involved in adipocyte differentiation, lipid storage and lipid metabolism. Bioinformatic analysis of cis-regulatory sequences of these genes revealed that their transcriptional regulators also overlapped, including EP300, HDAC2, CEBPB, CEBPD, FOXA1, and FOXA2. We also identified 15 and 46 genes that were dysregulated in the adipose tissue of DIO zebrafish and obese humans, respectively. Bioinformatics analysis identified EP300, HDAC2, and CEBPB as common transcriptional regulators for these genes. EP300 is a histone and lysyl acetyltransferase that modulates the function of histone and various proteins including CEBPB, CEBPD, FOXA1, and FOXA2. We demonstrated that adiposity in larval zebrafish was significantly reduced by C646, an inhibitor of EP300 that antagonizes acetyl-CoA. The reduction of adiposity by C646 was not significantly different from that induced by RSV or co-treatment of C646 and RSV. These results indicate that the inhibition of EP300 might be a common therapeutic mechanism between CR and RSV in adipose tissues of obese individuals.

ε-Viniferin, a resveratrol dimer, prevents diet-induced obesity in mice.

Red wines are thought to be one of the major dietary sources of trans-resveratrol. The beneficial effects of t-resveratrol against metabolic disorders have been well characterized, however, red wines also contain various resveratrol derivatives whose health benefits have not been completely elucidated. In this report, we investigated ε-viniferin, a resveratrol dimer, which is present at comparable concentrations to t-resveratrol in red wines, and has higher anti-adipogenesis activity in 3T3-L1 cells. In addition, ε-viniferin was more effective than t-resveratrol in its anti-obesity and anti-inflammatory effects in high-fat diet fed mice. These results suggested ε-viniferin may be one of the active ingredients against metabolic disorders in red wines, in addition to t-resveratrol.

Analysis of Rotundone in Japanese Syrah Grapes and Wines using Stir Bar Sorptive Extraction (SBSE) with Heart-Cutting Two-Dimensional GC-MS

Rotundone, an oxygenated sesquiterpene, is responsible for the peppery aroma in wines. We developed an effective and simple method to determine trace levels of rotundone in grapes and wines. The method uses stir bar sorptive extraction (SBSE) with heart-cutting two-dimensional gas chromatography/mass spectrometry without a pre-extraction process such as solid-phase extraction. The limits of detection and quantitation for rotundone were 2.1 and 6.5 ng/kg in grape and 2.4 and 7.2 ng/L in wine. The recoveries of rotundone from Cabernet Sauvignon grape and wine were 106% (spiked with 120 ng/kg for grape) and 96% (spiked with 120 ng/L for wine), respectively. We demonstrated that rotundone concentrations in Syrah grapes and wines from Ueda were higher (up to 2342 ng/kg and 232 ng/L, respectively) than the maximal rotundone concentrations in Australian Syrah grapes and wines. Findings indicate that some environmental factors in Japan may promote rotundone accumulation in Syrah grapes, and underscores the importance of studying regional characteristics of rotundone accumulation in Syrah grapes.

Impact of Harvest Timing on the Concentration of 3-Mercaptohexan-1-ol Precursors in Vitis vinifera Berries

The effect of harvest timing on the concentration of flavor precursor 3-mercaptohexan-1-ol (flavor precursor 3MH) in Vitis vinifera grape berries was investigated. The accumulation of flavor precursor 3MH in Chardonnay grape increased in early morning and decreased during the day. The daily fluctuation of flavor precursor 3MH accumulation in grape was common to several grape cultivars irrespective of the sampling year. These results suggested that early morning harvest might yield higher concentrations of 3MH and 3-mercaptohexyl acetate in wines than daytime harvest. These findings could contribute to improving viticultural and enological practices with the aim of producing aromatic wines.

Effects of Polysaccharide Derived from Black Currant on Relieving Clinical Symptoms of Japanese Cedar Pollinosis: A Randomized Double-Blind, Placebo-Controlled Trial

We investigated the efficacy of the polysaccharide derived from black currant, named cassis polysaccharide (CAPS), for inhibiting Japanese cedar pollinosis symptoms and improving quality of life by a randomized double-blind, placebo-controlled trial in 2006. A total of 28 subjects were enrolled in the study, and 10 subjects in each group completed the trial. Although there was no significant difference between the CAPS and placebo group in the weekly mean value of any symptom in the daily symptom diary at any time, a smaller degree of final symptom aggravation was found in the CAPS group. Significant aggravation of the score was finally observed in the placebo group with inferior conch swelling and with sneezing, itchy nose, itchy eye and watery eye in the Japan rhino-conjunctivitis quality of life questionnaire assessment, while the changes observed in the CAPS group were not significant. In conclusion, our findings clearly indicate that CAPS would be useful as a food supplement in assisting the treatment of Japanese cedar pollinosis.