Tyrosinase inhibitor from black rice bran

The inhibitor of tyrosinase activity in black rice bran was investigated. The methanol extract from black rice bran was re-extracted with hexane, chloroform, ethyl acetate, or water. The ethyl acetate extract had the most potent inhibition against tyrosinase activity by 80.5% at a concentration of 0.4 mg/mL. Inhibitory compound in the ethyl acetate fraction was isolated by silica gel column chromatography, and identified as protocatechuic acid methyl ester (compound 1) by GC, GC-MS, IR, and 1H and 13C NMR spectroscopy. Compound 1 inhibited 75.4% of tyrosinase activity at a concentration of 0.50 micromol/mL. ID(50) (50% inhibition dose) value of compound 1 was 0.28 micromol/mL. To study the structure-activity relationship, protocatechuic acid (2), vanillic acid (3), vanillic acid methyl ester (4), isovanillic acid (5), isovanillic acid methyl ester (6), veratric acid (7), and veratric acid methyl ester (8) were also assayed.     

A hairy root culture of melon produces aroma compounds

Musk melon is the favorite fruit with a high market value in Japan, and the fragrance is one of the major factors determining the fruit quality of melon. In this study, mutant melon hairy roots which had been induced by means of the T-DNA insertion mutagenesis were found to produce volatile compounds with the fruity fragrance of mature melon. The volatile compounds were extracted and identified by GLC-mass spectrometry. Some essential oils such as (Z)-3-hexenol, (E)-2-hexenal, 1-nonanol, and (Z)-6-nonenol were stably synthesized by these hairy roots despite the increased number of subcultures. The productivity of these compounds by the best hairy root line was shown to be considerably higher than naturally ripened melon fruits.     

Identification of an antioxidant,ethyl protocatechuate,in peanut seed testa

The antioxidant activity and identification of the antioxidant component of peanut seed testa were investigated. The antioxidant activity of peanut seed testa was studied in the linoleic acid model system by using the ferric thiocyanate method. Among the five organic solvent extracts, the ethanolic extracts of peanut seed testa (EEPST) produced higher yields and stronger antioxidant activity than other organic solvent extracts. EEPST was separated into 17 fractions on silica gel column chromatography. Fraction 17, which showed the largest yield and significant antioxidant activity, was separated by thin-layer chromatography. Four major antioxidative subfractions were present. Subfraction 17-2 was found to be effective in preventing oxidation of linoleic acid. This subfraction was further fractionated and isolated and characterized by UV, MS, IR, and (1)H NMR techniques. The active compound was identified as ethyl protocatechuate (3,4-dihydroxybenzoic acid ethyl ester).     

Loss of ripening capacity of pawpaw fruit with extended cold storage

The fruit ripening traits of pawpaw [ Asimina triloba (L.) Dunal] were examined after harvest and after cold storage at -2, 2, 4, and 6 degrees C for up to 12 weeks. Generally, fruits stored at 2-4 degrees C for 4 weeks ripened normally, but those stored at -2 degrees C did not ripen normally, those stored at 6 degrees C were overripe, and by 6-8 weeks those stored at 2-4 degrees C had a lower respiration rate and ethylene production, lower firmness, and lower pH than fruit cold-stored for 4 weeks or less. These changes, and the occasional development of brown discoloration in the pulp once the fruits were moved back to room temperature, were evidence of chilling injury by 6 weeks. After harvest and through 4 weeks of cold storage, the main volatile compounds produced by fruit were methyl and ethyl octanoates and hexanoates. Volatile production significantly increased >5-fold in fruit ripened for 72 h after harvest or after removal from up to 4 weeks of cold storage. Fruit cold-stored for 6 weeks or more produced fewer total volatiles and esters but increased levels of such off-flavor compounds as ethyl acetate, ethyl propionate, and hexanoic and decanoic acids. Alcohol acyltransferase (AAT) activity declined in cold-stored fruit but was not correlated with either total volatile production or total ester production. Alcohol dehydrogenase activity did not change during ripening after harvest or cold storage. Lipoxygenase activity was highest just after harvest or after 2 weeks of cold storage, but was low by 4 weeks. Thus, ripening of pawpaw fruit seems to be limited to 4 weeks at 2-4 degrees C with loss of ability to continue ripening and chilling injury symptoms evident at colder temperatures and after longer periods of cold storage.     

In vitro and in vivo stability of caffeic acid phenethyl ester, a bioactive compound of propolis

The in vitro biochemical stability of caffeic acid phenethyl ester in rat and human plasma was investigated and compared with the stability of other caffeic acid esters (chlorogenic acid and rosmarinic acid). The incubation of the compounds in rat plasma for up to 6 h showed that caffeic acid phenethyl ester, but not the other compounds, was hydrolyzed, whereas human plasma did not affect the stability of all the assayed compounds. The products in rat plasma were caffeic acid and an unknown compound, which was identified by mass spectrometry as caffeic acid ethyl ester, produced by transesterification in the presence of ethanol used as vehicle for standard compounds. Specific inhibitors of different plasma esterases allowed the identification of a carboxylesterase as the enzyme involved in the metabolism of caffeic acid phenethyl ester. The oral administration in rats of caffeic acid phenethyl ester in the presence of both ethanol and 2-(2-ethoxyethoxy)ethanol gave rise to a dramatic increase of caffeic acid, as well as low levels of caffeic acid phenethyl ester, caffeic acid ethyl ester, and caffeic acid 2-(2-ethoxyethoxy)ethyl ester, in urine collected within 24 h after treatment. These results suggest that caffeic acid phenethyl ester is hydrolyzed also in vivo to caffeic acid as the major metabolite and that its biological activities should be more properly assayed and compared with those of caffeic acid, its bioactive hydrolysis product. Moreover, alcohols should be carefully used in vivo as solvents for caffeic acid phenethyl ester, since they can give rise to new bioactive caffeic acid esters.     

不同氮素水平对甜瓜芳香物质和营养品质的影响

以京玉五号为试材,利用GC-MS测定法,在溶液培养条件下研究了不同氮素供应水平对甜瓜芳香物质和营养品质的影响。结果表明,在不同氮素水平条件下,甜瓜的芳香物质的种类和含量差异显著。乙酸-2-甲基丙基酯、乙酸丁酯和2-甲基丁酸乙酯等特征芳香物质的含量随着氮素水平的升高先升高后下降,在100%的氮素处理水平下,甜瓜的特征芳香物质含量相对较高。随着氮素水平的升高,甜瓜的折光糖含量、Vc、可溶性糖和糖酸比等营养品质表现下降的趋势。研究结果表明,在本试验条件下,200.72～401.44 mg/L氮素是甜瓜优质高产栽培适宜的氮素水平。     

Constituents of Asparagus officinalis evaluated for inhibitory activity against cyclooxygenase-2

As part of a project directed toward the discovery of new cancer chemopreventive agents from plants, two new natural products, asparagusic acid anti-S-oxide methyl ester (1) and asparagusic acid syn-S-oxide methyl ester (2), a new acetylenic compound, 2-hydroxyasparenyn [3',4'-trans-2-hydroxy-1-methoxy-4-[5-(4-methoxyphenoxy)-3-penten-1-ynyl]-benzene] (3), as well as eleven known compounds, asparenyn (4), asparenyol (5), (+/-)-1-monopalmitin (6), ferulic acid (7), 1,3-O-di-p-coumaroylglycerol (8), 1-O-feruloyl-3-O-p-coumaroylglycerol (9), blumenol C, (+/-)-epipinoresinol, linoleic acid, 1,3-O-diferuloylglycerol, and 1,2-O-diferuloylglycerol, were isolated from an ethyl acetate-soluble fraction of the methanol extract of the aerial parts of Asparagus officinalis (Asparagus), using a bioassay based on the inhibition of cyclooxygenase-2 to monitor chromatographic fractionation. The structures of compounds 1-3 were elucidated by 1D- and 2D-NMR experiments ((1)H NMR, (13)C NMR, DEPT, COSY, HMQC, HMBC and NOESY). All the isolates were evaluated for their inhibitory effects against both cyclooxygenase-1 and -2, with the most active compound being linoleic acid.     

Functionality of bioactive compounds in Brazilian strawberry (Fragaria x ananassa Duch.) cultivars: evaluation of hyperglycemia and hypertension potential using in vitro models

Fruits of seven fully ripened strawberry cultivars grown in Brazil (Dover, Camp Dover, Camarosa, Sweet Charlie, Toyonoka, Oso Grande, and Piedade) were evaluated for total phenolics, antioxidant activity based on DPPH radical scavenging assay, and functionality such as inhibition of alpha-amylase, alpha-glucosidase, and angiotensin I-converting enzyme (ACE) relevant for potentially managing hyperglycemia and hypertension. The total phenolics content ranged from 966 to 1571 microg of gallic acid/g of fruit fresh weight for Toyonoka and Dover, respectively. No correlation was found between total phenolics and antioxidant activity. The major phenolic compounds in aqueous extracts of strawberries were ellagic acid, quercetin, and chlorogenic acid. Strawberries had high alpha-glucosidase inhibitory activity. However, alpha-amylase inhibitory activity was very low in all cultivars. This suggested that strawberries could be considered as a potential dietary source with anti-hyperglycemic potential. The evaluated cultivars had no significant ACE inhibitory activity, reflecting low anti-hypertensive potential.     

Biochemical and catalytic properties of three recombinant alcohol acyltransferases of melon. sulfur-containing ester formation, regulatory role of CoA-SH in activity, and sequence elements conferring substrate preference

Alcohol acyltransferases (AAT) play a key role in the biosynthesis of ester aroma volatiles in fruit. Three ripening-specific recombinant AATs of cantaloupe Charentais melon fruit (Cm-AAT1, Cm-AAT3, and Cm-AAT4) are capable of synthesizing thioether esters with Cm-AAT1 being by far the most active. All proteins, as well as AAT(s) extracted from melon fruit, are active as tetramers of around 200 kDa. Kinetic analysis demonstrated that CoA-SH, a product of the reaction, is an activator at low concentrations and an inhibitor at higher concentrations. This was confirmed by the addition of phosphotransacetylase at various concentrations, capable of modulating the level of CoA-SH in the reaction medium. Site-directed mutagenesis of some amino acids that were specific to the Cm-AAT sequences into amino acids that were consensus to other characterized AATs greatly affected the selectivity of the original protein and the number of esters produced.     

不同氮素水平对棚栽草莓果实芳香成分的影响

采用溶剂萃取法提取经不同氮素水平处理后的草莓果实芳香成分,进行气相色谱质谱联用仪(GC/MS)分析。结果表明,高量氮肥、中量氮肥、低量氮肥、不施氮肥等4个处理分别检测出28种、25种、29种、28种芳香成分,各占总峰面积的93.55%,93.40%,92.79%,83.07%。各处理成熟草莓果实特征芳香成分中丁酸甲酯、丁酸乙酯、己酸甲酯、己酸乙酯、2,5-二甲基-4-羟基-3(2H)-呋喃酮、2,5-二甲基-4-甲氧基-3(2H)-呋喃酮等的相对含量存在差异;不同氮素水平对成熟草莓果实风味品质影响明显。     

Fruit maturity and juice extraction influences ellagic acid derivatives and other antioxidant polyphenolics in muscadine grapes

Polyphenolic compounds including ellagic acid, ellagic acid derivatives, and anthocyanins were characterized and quantified by novel chromatographic conditions in eight muscadine grape (Vitis rotundifolia) cultivars and evaluated for antioxidant capacity as influenced by two ripening stages and their location within the fruit (skin, pulp, and juice). All polyphenolics generally increased as fruit ripened and the highest concentrations were located in the skins. Free ellagic acid, ellagic acid glycosides, and total ellagic acid ranged from 8 to 162, 7 to 115, and 587 to 1900 mg/kg, respectively, in the skin of ripe grapes. Hot-pressed juices contained considerably lower polyphenolic concentrations than were present in whole grapes. Five anthocyanidins were present in each cultivar in variable concentrations (delphinidin > petunidin > malvidin + peonidin > cyanidin). Antioxidant capacity was appreciably influenced by cultivar, maturity, and location in the fruit with good correlations to soluble phenolics found in both methanolic and ethyl acetate extracts (r = 0.83 and 0.92, respectively).     

Mumefural, citric acid derivative improving blood fluidity from fruit-juice concentrate of Japanese apricot (Prunus mume Sieb. et Zucc).

The effects of food components on blood fluidity were studied by in vitro assay using a dedicated microchannel instrument for model capillaries. We found that the fruit-juice concentrate of the Japanese apricot (Prunus mume Sieb. et Zucc), a traditional Japanese food, markedly improved the fluidity of human blood. Using HPLC, we isolated the active compounds and characterized them using UV, MS, IR, and NMR. They included a novel compound, 1-[5-(2-formylfuryl)methyl] dihydrogen 2-hydroxypropane-1,2, 3-tricarboxylate (mumefural), and a related compound, 5-hydroxymethyl-2-furfural (HMF). Mumefural markedly improved blood fluidity in all subjects, while HMF worked differently in different individuals. The flow rate of blood spiked with mumefural or HMF was compared to that of the two predominant organic acids in the fruit. Citric acid, malic acid, and furfuryl alcohol also improved fluidity in all subjects. The activity of P. mume is derived from not only artifacts produced during thermal processing, such as mumefural, but also from endogenous organic acids.     

Antioxidant, sugar, mineral, and phytonutrient concentrations across edible fruit tissues of orange-fleshed honeydew melon (Cucumis melo L.)

Orange-fleshed, non-netted honeydew ( Cucumis melo L.) is a relatively new melon in the marketplace and has shown a lot of potential as an alternative to netted muskmelons (cantaloupes), which are often prone to surface contamination by enteric bacteria. Orange-fleshed honeydew is a cross between orange-fleshed cantaloupe and non-netted, green-fleshed honeydew. This glasshouse study investigated the nutritional profile (phytonutrient and sugar contents) in different tissues of mature orange-fleshed honeydew melon fruit. The equatorial mesocarp of ripe fruit was segmented into hypodermal (subpeel), outer, middle, and inner (near the seed cavity) tissues and then assayed for total sugars, mineral nutrients, phytonutrients, total proteins, and enzymatic antioxidant activities. The concentrations of soluble solids, sucrose, total sugars, beta-carotene, and 5-methyltetrahydrofolic acid increased in an inward direction from the subpeel mesocarp tissues toward the seed cavity. The activities of ascorbate peroxidase, catalase, and superoxide dismutase also increased in an inward direction. The concentrations of calcium, iron, magnesium, manganese, and sodium all decreased in the inward direction. When expressed on a dry weight basis, the concentrations of ascorbic acid, boron, copper, fructose, glucose, phosphorus, potassium, and zinc were higher in the subpeel region compared to the inner mesocarp tissues, but the reverse was true when data were expressed on a fresh weight basis. These data reveal that there is considerable variation in sugars, minerals, and phytonutrients across the mesocarp regions and that expressing the data on a fresh or dry weight basis can alter interpretations of the nutritional significance and health benefits of fruit. The data also confirm that orange-fleshed honeydew melon can be a rich source of many human health-related nutrients.     

3种不同腌渍方式对腌渍黄瓜品质的影响

为改善低盐腌渍蔬菜品质下降的问题,本研究通过分析测定腌渍黄瓜的微观结构、质构性质、风味物质,研究低盐腌渍、复合腌制剂腌渍、高盐腌渍3种不同腌渍方式对腌渍黄瓜品质的影响。结果表明,高盐腌渍黄瓜和复合腌制剂腌渍黄瓜的质构及风味品质均优于低盐腌渍黄瓜。其中,高盐腌渍黄瓜的硬度略高于复合腌制剂腌渍黄瓜,氨基酸总量和有机酸总量差别不大;复合腌制剂腌渍黄瓜的硬度比低盐腌渍黄瓜高271 g;氨基酸、有机酸总量分别比低盐腌渍黄瓜样品高35.3和55.1 mg·100g^-1。 复合腌制剂腌渍黄瓜的质构、风味和感官等品质接近传统高盐腌渍黄瓜,优于低盐腌渍黄瓜。本研究结果为有效解决低盐腌渍使得黄瓜品质下降提供了一定的数据支持。     

Identification of aroma-active compounds in Jiashi muskmelon juice by GC-O-MS and OAV calculation

To identify aromatic compounds in Jiashi melon juice, gas chromatography-mass spectrometry-olfactometry (GC-MS-O) analysis was used. Odor activity values (OAVs) were also calculated on the basis of the qualitative and quantitative analysis of volatile compounds. Results showed that 42 volatiles were identified, among which 4 compounds, namely, diethyl carbonate, isophorone, 2-butoxyethyl acetate, and menthol, were identified or tentatively identified for the first time as volatiles in melon fruit. Twelve compounds, namely, (2E,6Z)-nona-2,6-dienal, (3Z,6Z)-nona-3,6-dien-1-ol, ethyl butanoate, ethyl 2-methylbutyrate, ethyl 2-methylpropanoate, (Z)-non-6-enal, (E)-2-nonenal, heptanal, methyl 2-methylbutyrate, nonanal, hexanal, and 2-methylpropyl acetate, were identified as the potent odorants of Jiashi melon juice by both OAV and detection frequency analysis (DFA). In addition, seven odorants were detected by all of the panelists and showed higher OAVs, indicating that DFA and OAV resulted in relatively similar "Jiashi" melon aroma patterns.     

Separation, characterization, and quantitation of phenolic acids in a little-known blueberry (Vaccinium arctostaphylos L.) fruit by HPLC-MS

The aim of this study was the qualitative and quantitative determination of free, ester, glycoside, and ester-bound phenolic acids in the blueberry (Vaccinium arctostaphylos L.) fruit. A method for the determination of the profile of phenolic acids of four different phenolic fractions in the fruit was developed using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Thirteen compounds (gallic, protocatechuic, p-hydroxybenzoic, m-hydroxybenzoic, gentisic, chlorogenic, p-coumaric, caffeic, ferulic, syringic, sinapic, salicylic, and trans-cinnamic acids) were identified and quantified in the berry. These experimental results showed that the predominant phenolic acid in the fruit of V. arctostaphylos is caffeic acid in free and insoluble ester-bound forms and p-coumaric acid in soluble ester and glycoside forms. Seven phenolic acids were identified as hydroxybenzoic acid derivatives (HBAs) and four as hydroxycinnamic acid derivatives (HCAs). Total content of HBAs and HCAs in the four phenolic fractions constituted 30.1 and 69.9% of the free, 27.9 and 72.1% of the ester, 24.7 and 75.3% of the glycoside, and 51.7 and 48.3% of the ester-bound forms, respectively. Total phenolics as the sum of individual phenolic acids identified is 698.5 ng/g of fresh weight (fw) for the free, 3399.2 ng/g of fw for the ester, 3522.1 ng/g of fw for the glycoside, and 3671.6 ng/g of fw for the ester-bound phenolic fractions. The present results were compared with reported levels of phenolic acids in the fruits of different Vaccinium species. These data suggest that the fruit can be considered as a potentially good dietary source of phenolic acids.     

Wild melon diversity in India (Punjab State)

We present here the first comprehensive genetic characterization of wild melon accessions from northern India. The genetic diversity among 43 wild melon accessions collected from the six agro-ecological regions of the Punjab State of India was assessed by measuring variation at 16 Simple Sequence Repeat (SSR) loci, morphological traits of plant habit and fruit morphological traits, two yield-associated traits, root nematode resistance and biochemical composition (ascorbic acid, carotenoids, titrable acidity). Variation among accessions was observed in plant habit and fruit traits and wild melon germplasm with high acidity and elevated carotenoid content and possessing resistance to Meloidogyne incognita was identified in the collection. A high level of genetic variability in wild melon germplasm was suggested by SSR analysis. Comparative analysis using SSRs of the genetic variability between wild melons from the north and other melons from the south and east regions of India and also reference accessions of cultivated melon from Spain, Japan, Korea, Maldives, Iraq and Israel, showed regional differentiation among Indian melon accessions and that Indian germplasm was not closely related to melon accessions from other parts of the world. A highly drought tolerant accession belonging to var. agrestis Naud. was also identified.     

Biosynthesis of strawberry aroma compounds through amino acid metabolism

The fate of amino acids in relation to aroma biogenesis was studied in strawberries using the in vitro growth approach. This fruit presented differences in the level of metabolization for different amino acids. Incubations of strawberries with L-isoleucine gave rise to an increase of fourteen compounds in this fruit aroma, either not detected previously or constituents of strawberry aroma. However, L-valine incubations did not provide a significant change in this fruit aroma. Strawberry feeding with L-isoleucine resulted in a 7-fold increase in the sum of 2-methylbutanoate esters, and a double production of 2-methylbutyl esters compared to those of control fruits. Around 94% of the ester increase corresponded to 2-methylbutanoates, with ethyl 2-methylbutanoate being the most representative compound (92%). On the other hand, among the 2-methylbutyl esters, comprising around 6% of total aroma volatiles increase, 2-methylbutyl acetate was the major compound (95%) arising from L-isoleucine strawberry feeding. The role of enzymatic activities within the amino acid metabolic pathway in strawberry fruits is discussed.     

Isolation, characterization and antifungal activity of major constituents of the Himalayan lichen Parmelia reticulata Tayl

Antifungal activity of hexane, ethyl acetate and methanol extracts of Parmelia reticulata was evaluated against soilborne pathogenic fungi, namely, Sclerotium rolfsii, Rhizoctonia solani, R. bataticola, Fusarium udum, Pythium aphanidermatum and P. debaryanum by poisoned food technique. Maximum antifungal activity was exhibited by hexane and ethyl acetate extracts against most of the test pathogens. Secondary metabolites, namely, (±)-isousnic acid, (±)-protolichesterinic acid, atranorin, evernyl, ethyl hematommate, ethyl orsellinate, methyl hematommate (3-formyl-2,4-dihydroxy-6-methylbenzoic acid methyl ester), 2-hydroxy-4-methoxy-3,6-dimethylbenzoic acid, 1-hydroxy-3,6-dimethoxy-8-methyl-xanthen-9-one, baeomycesic acid and salazinic acid, were isolated from the above extracts and identified by 1H NMR, 13C NMR and mass spectroscopic methods. When these metabolites were tested for antifungal activity against test pathogens, maximum antifungal activity was exhibited by (±)-protolichesterinic acid against R. solani (ED50=23.09 μg mL(-1)) and P. debaryanum (ED50=16.07 μg mL(-1)) and by atranorin against S. rolfsii (ED50=39.70 μg mL(-1)). The antifungal activity of protolichesterinic acid was found to be comparable to that of hexaconazole, a commercial fungicide.     

Identification of triterpene hydroxycinnamates with in vitro antitumor activity from whole cranberry fruit (Vaccinium macrocarpon)

Bioactivity-guided fractionation of cranberry fruit was used to determine the identity of triterpenoid esters from Vaccinium macrocarpon, which inhibit tumor cell growth and may play a role in cancer prevention. In our previous study, a fraction from whole fruit exhibited tumor cell growth inhibition in vitro. The major components of this fraction were isolated by chromatographic separation of ethyl acetate extracts, purified by semipreparative HPLC, and identified by NMR as cis- (1) and trans- (2) isomers of 3-O-p-hydroxycinnamoyl ursolic acid. These triterpenoid esters have not been previously reported in Vaccinium fruit. Bioassay of the purified triterpene cinnamates in tumor cell lines in vitro showed slightly greater activity of compound 1 in most cell lines, with GI(50) values of approximately 20 microM in MCF-7 breast, ME180 cervical and PC3 prostate tumor cell lines. Quercetin was slightly less active than 1, while cyanidin-3-galactoside exhibited much lower cytotoxicity, with GI(50) greater than 250 microM in all cell lines. Phenylboronic acid (3) was also isolated from the fruit but showed insignificant antitumor activity.