Lemon peel and Limoncello liqueur: A proteomic duet

Combinatorial peptide ligand libraries (CPLLs) have been adopted for investigating the proteomes of lemon peels and pulp, of a home-made alcoholic infusion of peels and of a very popular Italian liqueur called “Limoncello”, stated to be an infusion of the flavedo (the outer, yellow skin of lemons). The aim of this study was not only to perform the deepest investigation so far of the lemon peel proteome but also to assess the genuineness of the commercial liqueur via a three-pronged attack. First, different extraction techniques have been used for the characterization of the peel (and additionally of the pulp) proteome, secondly a home-made infusion has been analysed and finally the proteome of the commercial drink was checked. The peel (the flavedo, not the underlying layer called albedo) proteome has been evaluated via prior capture with CPLLs at different pH values (2.2 and 7.2). Via mass spectrometry analysis of the recovered fractions, after elution of the captured populations in 4% boiling SDS, we could identify a total of 1011 unique gene products in the peel extracts and 674 in the pulp, 264 proteins in the home-made infusion and just 8 proteins (and protein fragments), together with 12 peptides, in one Italian Limoncello produced in the Sorrento Region, thus proving the genuineness of this product. On the contrary, cheaper Limoncellos were devoid of any protein/peptide, casting doubts on their production from vegetable extracts. This could be the starting point for investigating the genuineness and natural origin of commercial drinks in order to protect consumers from adulterated products.     

Production and optimization of polygalacturonase from mango (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.) peel using <Emphasis Type="Italic">Fusarium moniliforme</Emphasis> in solid state fermentation

Mango peel is one of the major wastes from fruit processing industries, which poses considerable disposal problems and ultimately leads to environmental pollution. The objective of the current research was to determine the significant parameters on the production of polygalacturonase from mango peel which is a major industrial waste. Solid state culture conditions for polygalacturonase production by Fusarium moniliforme from dried mango peel powder were optimized by Taguchi’s L-18 orthogonal array experimental design methodology. Eight fungal metabolic influencing variables, viz. temperature, mango peel, inoculum, peptone, ammonium nitrate (NH₄NO₃₎, magnesium sulphate (MgSO₄), zinc sulphate (ZnSO₄) and potassium dihydrogen phosphate (KH₂PO₄) were selected to optimize polygalacturonase production. The optimized parameters composed of temperature (30°C), mango peel (6.5%, g, w/v), inoculum (8%, ml, v/v), peptone (1%, g, w/v), NH₄NO₃ (0.60%, g, w/v), MgSO₄ (0.05%, g, w/v), ZnSO₄ (0.06%, g, w/v) and KH₂PO₄ (0.4%, g, w/v). Based on the influence of interaction of fermentation components of fermentation, these could be classified as the least significant and the most significant at individual and interaction levels. The temperature, inoculum level, mango peel substrate and KH₂PO₄ showed maximum production impact at optimized conditions. From the optimized conditions the polygalacturonase activity was maximized to 43.2 U g⁻¹.     

According to the CPLL proteome sheriffs,not all aperitifs are created equal!

Combinatorial peptide ligand libraries (CPLLs) have been adopted for investigating the proteome of a popular aperitif in Northern Italy, called “Amaro Branzi”, stated to be an infusion of a secret herbal mixture, of which some ingredients are declared on the label, namely Angelica officinalis, Gentiana lutea and orange peel, sweetened by a final addition of honey. In order to assess the genuineness of this commercial liqueur, we have prepared extracts of the three vegetable ingredients, assessed their proteomes, and compared them to the one found in the aperitif. The amaro's proteome was identified via prior capture with CPLLs at two different pH values (2.2 and 4.8). Via mass spectrometry analysis of the recovered fractions, after elution of the captured populations in 4% boiling SDS, we could confirm the presence of the following: six proteins originating from honey, 11 from orange peels, 29 from G. lutea and 46 from A. officinalis (including shared species), plus 33 species which could not be attributed to the other secret ingredients, due to paucity of genomic data on plant proteins, for a total of 93 unique gene products (merging shared proteins). This fully confirmed the genuineness of the product. Considering that most of these species could be present in trace amounts, undetectable by conventional techniques, the CPLL methodology, due to its ability to enhance the signal of trace components up to 3 to 4 orders of magnitude, could represent a powerful tool for investigating the genuineness and natural origin of commercial beverages in order to protect consumers from adulterated products.     

八种水果中的多酚含量及其抗氧化性

测定了苹果、石榴、橄榄、芒果、香蕉、菠萝、葡萄和龙眼的壳、肉及核中的多酚含量及其抗氧化性。以70%丙酮(v/v)为提取溶剂,室温下超声波辅助浸提样品后得到提取液。采用普鲁士兰(Prussian blue)法测定了提取液中多酚和单宁的含量,利用FRAP法测定其抗氧化性。结果表明:提取液中多酚和单宁的含量均与其抗氧化性成正相关关系;石榴、橄榄、芒果、葡萄和龙眼等水果的壳与核有望成为天然抗氧化剂的新来源。     

Lipofuscin like compound in mango

Thin layer chromatographic separation of chloroform-methanol extracts of mango on silica gel revealed a fluorescent substance in mango peel and pulp. The compound had fluorescence spectrum similar to that of lipofuscin, the age pigment of animal tissues and was found to be water insoluble and stable to ultraviolet irradiation. The fluorescent material appeared to be a lipoprotein.     

杧果采后生理研究

适期采收的果．采收时呼吸速率为２３．２ｍｇＣＯ２·ｋｇ（－１）ｈ（－１）果皮深绿色．果肉硬度＞３０Ｐ／ｃｍ２．置室温下（３０℃）８ｄ出现呼吸高峰,速率１３２．９ｍｇＣＯ２·ｋｇ（－１）ｈ（－１）,果皮的颜色黄绿各半．果肉硬度８—９Ｐ／ｃｍ２。采后１２—１４ｄ果实达到完熟,果皮金黄色,硬度＜３Ｐ／ｃｍ２,呼吸速率下降到３９—４５ｍｇＣＯ２·ｋｇ（－１）ｈ（－１）。试验结果还表明．可溶性糖、还原糖、非还原糖、固形物等含量．均在采后第５天明显增高,果皮从深绿转为淡绿色。可溶性糖在采后第１４天增至最高．达９．９２％,此时可滴定酸度下降至０．１４％．ＰＨ值升至６．１５,此时为果实的最佳供食状态．生产上可据此为制定果贮藏措施提供依据．     

Artichoke and Cynar liqueur: Two (not quite) entangled proteomes

Combinatorial peptide ligand libraries (CPLLs) have been adopted to investigate the proteome of artichoke extracts, of a home-made alcoholic infusion and of the Italian Cynar liqueur. The aim of study was not only to perform the deepest investigation so far of the artichoke proteome but also to assess the genuineness of the commercial aperitif via a three-pronged attack. First, different extraction techniques have been used for the characterization of the artichoke's proteome, secondly a home-made infusion has been analyzed and finally the proteome of the commercial drink was checked. The artichoke proteome has been evaluated via prior capture with CPLLs at four different pH (2.2, 4.0, 7.2 and 9.3) values. Via mass spectrometry analysis of the recovered fractions, after elution of the captured populations in 4% boiling SDS, we could identify a total of 876 unique gene products in the artichoke extracts, 18 in the home-made infusion and no proteins at all in the Italian Cynar liqueur, casting severe doubts on the procedure stated by the manufacturer (that should be made by an infusion of artichoke leaves plus thirteen different herbs). This could be the starting point for investigating the genuineness and natural origin of commercial drinks in order to protect consumers from adulterated products.     

Effect of bovicin HC5 on growth and spore germination of Bacillus cereus and Bacillus thuringiensis isolated from spoiled mango pulp

AIMS: To use bovicin HC5 to inhibit predominant bacteria isolated from spoiled mango pulp. METHODS AND RESULTS: Bovicin HC5 and nisin were added to brain heart infusion (BHI) medium (40-160 AU ml(-1)) or mango pulp (100 AU ml(-1)) and the growth of Bacillus cereus and Bacillus thuringiensis was monitored. Cultures treated with bovicin HC5 or nisin showed longer lag phases and grew slower in BHI medium. Bovicin HC5 and nisin were bactericidal and showed higher activity in mango pulp at acidic pH values. To determine the effect on spore germination and D values, mango pulp containing bovicin HC5 was inoculated with 10(6) and 10(9) spores per ml(-1), respectively, from each strain tested. Bovicin HC5 reduced the outgrowth of spores from B. cereus and B. thuringiensis, but thermal sensitivity was not affected. CONCLUSIONS: Bovicin HC5 was bactericidal against B. cereus and B. thuringiensis isolated from spoiled mango pulp. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacillus cereus and B. thuringiensis had not been previously isolated from spoiled mango pulp and bovicin HC5 has the potential to inhibit such bacteria in fruit pulps.     

Usefulness of manufactured tomato extracts in the diagnosis of tomato sensitization: Comparison with the prick-prick method

Background

Commercial available skin prick test with fruits can be negative in sensitized or allergic patients due to a reduction in biological activity during the manufacturing process. Prick-prick tests with fresh foods are often preferred, but they are a non-standardized procedure. The usefulness of freeze-dried extracts of Canary Islands tomatoes, comparing the wheal sizes induced by prick test with the prick-prick method in the diagnosis of tomato sensitization has been analyzed. The objective of the study was to assess the potential diagnostic of freeze-dried extracts of Canary Islands tomatoes, comparing the wheal sizes induced by prick test with the prick-prick method.

Methods

Two groups of patients were analyzed: Group I: 26 individuals reporting clinical symptoms induced by tomato contact or ingestion. Group II: 71 control individuals with no symptoms induced by tomato: 12 of them were previously skin prick test positive to a tomato extract, 39 were atopic and 20 were non-atopic. All individuals underwent prick-prick with fresh ripe peel Canary tomatoes and skin prick tested with freeze-dried peel and pulp extracts obtained from peel and pulp of Canary tomatoes at 10 mg/ml. Wheal sizes and prick test positivity (≥ 7 mm²) were compared between groups.

Results

In group I, 21 (81%) out of 26 patients were prick-prick positive. Twenty patients (77%) had positive skin prick test to peel extracts and 12 (46%) to pulp extracts. Prick-prick induced a mean wheal size of 43.81 ± 40.19 mm² compared with 44.25 ± 36.68 mm² induced by the peel extract (Not significant), and 17.79 ± 9.39 mm² induced by the pulp extract (p < 0.01). In group II, 13 (18%) out of 71 control patients were prick-prick positive. Twelve patients (all of them previously positive to peel extract) had positive skin prick test to peel and 3 to pulp. Prick-prick induced a mean wheal size of 28.88 ± 13.12 mm² compared with 33.17 ± 17.55 mm² induced by peel extract (Not significant), and 13.33 ± 4.80 mm² induced by pulp extract (p < 0.05 with peel extract and prick-prick).

Conclusion

Canary peel tomato extract seems to be as efficient as prick-prick tests with ripe tomatoes to diagnose patients sensitized to tomato. The wheal sizes induced by prick-prick and peel extracts were very similar and showed a high correlation coefficient.     

Role of Antifungal Gallotannins,Resorcinols and Chitinases in the Constitutive Defence of Immature Mango (Mangifera indica L.) against Colletotrichum gloeosporioides

Conidia of Colletotrichum gloeosporioides germinate and form infection hyphae on inoculated, immature mango but remain quiescent until fruit ripening. Antifungal resorcinols have previously been implicated for quiescence of C. gloesoporioides and Alternaria alternata on mango. This study revealed the presence of a mixture of several gallotannins with glycosidic linkages, including 1,2,3,4,6‐penta‐O‐galloyl‐β‐D‐glucopyranose, with significant antifungal activity in the unripe mango fruit peel. Gallotannin antifungal activity was greater in a cultivar resistant (295.8 mm² inhibition) to anthracnose than in a susceptible (148.4 mm² inhibition) cultivar. In both, the activity decreased with ripening but the decrease was 10% less in the resistant cultivar. Three recorcinols, 5‐pentadecylresorcinol, 5‐(12‐cis‐heptadecenyl)resorcinol, AR 21 and another resorcinol derivative were present in the unripe fruit peel and all declined during ripening, more significantly the 5‐(12‐cis‐heptadecenyl)resorcinol and AR 21. Mango latex, when drained out, separates into an oily and aqueous phase. The aqueous phase showed significant chitinase activity and the ability to digest conidia of C. gloeosporioides. The oily phase has previously been reported to contain resorcinols. Draining fruits of latex soon after harvest resulted in greater incidence and severity of anthracnose at ripe stage. Chitinase activity was less in the peel of fruits from which latex was drained. The evidence suggests that the resistance of unripe mango to C. gloeosporioides is because of an elaborate constitutive defence system comprising antifungal resorcinols, gallotannins and chitinases.     

Identification of olive (Olea europaea) seed and pulp proteins by nLC-MS/MS via combinatorial peptide ligand libraries

Different types of extraction protocols are described for identifying proteins in seed and pulp of olive (Olea europea), by employing both conventional extraction methods and capture with ProteoMiner as well as with in house-made combinatorial peptide ligand libraries (HM-CPLLs) at pH 7.4 and at pH 2.2. Thanks to the use of CPLLs, able to dramatically amplify the signal of low-abundance species, a quite large number of compounds has been indeed identified: 61 in the seed (vs. only four reported in current literature) and 231 in the pulp (vs. 56 described so far), the deepest investigation up to the present of the olive proteome. In the seed, it highlights the presence of seed storage proteins, oleosins and histones. In the pulp, the allergenic thaumatin-like protein (Ole e 13) was confirmed, among the other 231, as the most abundant protein in the olive pulp. The present research has also been undertaken with the aim of identifying proteins in olive oil and ascertaining the relative contribution of seed and pulp proteins in their presence, if any, in oils.     

SENSORY QUALITY STUDIES ON MANGO AROMA CONCENTRATE DURING STORAGE

Aroma concentrates from Alphonso and Totapuri mango pulps were prepared and the aroma stripped pulps were concentrated two folds. Ready to serve beverages were prepared from fresh mango pulps and pulp concentrates by adding different levels of aroma concentrates. Sensory evaluation of beverages showed that in the case of Alphonso mango addition of 50%, 75% and 100% aroma concentrate gave comparable quality while with Totapuri only 50% level showed comparable quality. The aroma stripped pulp concentrates were canned as such and after mixing with 50% aroma concentrates and stored at 25 × 5°C. Aroma concentrates were bottled and stored at 2°C. At different intervals of storage, beverages were prepared and evaluated. The results showed that throughout the storage period, beverages prepared from mango concentrates to which 50% aroma was added before canning was superior to those prepared from aroma stripped canned pulp concentrates to which aroma was added at the time of preparation of beverage indicating the effect of mango pulp on aroma stability.     

Seasonal pattern in food gathering of the weaver ant Oecophylla longinoda (Hymenoptera: Formicidae) in mango orchards in Benin

Prey capture and food scavenging activities of Oecophylla longinoda were monitored through regular weekly samplings during two consecutive years (2009–2010) in a large mango orchard of the Borgou Department of Benin, West Africa, a main mango production area located in the Sudan agro-ecological zone. In both years, interspecific competition with other ants occurred mainly during the dry season (January to March) resulting in increased captures of Formicinae, Myrmicinae and Ponerinae. More prey was caught during the rainy season (end April to end October) than during the rest of the year, with Diptera and Coleoptera prey peaking in May and June, respectively, along with the mango season. As insect prey quickly decreased during November to December weaver ants increasingly collected seeds and plant debris. A total of 241 species of insects were captured including 61 species (25.3%) associated with mango and a few with cashew, among which 48 mango pest species (78.7% of species associated with mango tree). Only five species (2.1%) of beneficial insects were captured. It is concluded that the presence of O. longinoda colonies is beneficial to perennial tree cropping systems such as mango and cashew.     

Sequence diversity and differential expression of major phenylpropanoid-flavonoid biosynthetic genes among three mango varieties

Background

Mango fruits contain a broad spectrum of phenolic compounds which impart potential health benefits; their biosynthesis is catalysed by enzymes in the phenylpropanoid-flavonoid (PF) pathway. The aim of this study was to reveal the variability in genes involved in the PF pathway in three different mango varieties Mangifera indica L., a member of the family Anacardiaceae: Kensington Pride (KP), Irwin (IW) and Nam Doc Mai (NDM) and to determine associations with gene expression and mango flavonoid profiles.

Results

A close evolutionary relationship between mango genes and those from the woody species poplar of the Salicaceae family (Populus trichocarpa) and grape of the Vitaceae family (Vitis vinifera), was revealed through phylogenetic analysis of PF pathway genes. We discovered 145 SNPs in total within coding sequences with an average frequency of one SNP every 316 bp. Variety IW had the highest SNP frequency (one SNP every 258 bp) while KP and NDM had similar frequencies (one SNP every 369 bp and 360 bp, respectively). The position in the PF pathway appeared to influence the extent of genetic diversity of the encoded enzymes. The entry point enzymes phenylalanine lyase (PAL), cinnamate 4-mono-oxygenase (C4H) and chalcone synthase (CHS) had low levels of SNP diversity in their coding sequences, whereas anthocyanidin reductase (ANR) showed the highest SNP frequency followed by flavonoid 3’-hydroxylase (F3’H). Quantitative PCR revealed characteristic patterns of gene expression that differed between mango peel and flesh, and between varieties.

Conclusions

The combination of mango expressed sequence tags and availability of well-established reference PF biosynthetic genes from other plant species allowed the identification of coding sequences of genes that may lead to the formation of important flavonoid compounds in mango fruits and facilitated characterisation of single nucleotide polymorphisms between varieties. We discovered an association between the extent of sequence variation and position in the pathway for up-stream genes. The high expression of PAL, C4H and CHS genes in mango peel compared to flesh is associated with high amounts of total phenolic contents in peels, which suggest that these genes have an influence on total flavonoid levels in mango fruit peel and flesh. In addition, the particularly high expression levels of ANR in KP and NDM peels compared to IW peel and the significant accumulation of its product epicatechin gallate (ECG) in those extracts reflects the rate-limiting role of ANR on ECG biosynthesis in mango.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1784-x) contains supplementary material, which is available to authorized users.     

Analysis and characterization of anthocyanins and carotenoids in Japanese blue tomato

Tomato (Solanum lycopersicum) is rich in anthocyanins, which are polyphenolic pigments. This study aimed to analyze and characterize the anthocyanin composition in cultivated blue tomato in Japan. The extracts of peel, seed, and pulp of tomatoes were purified following which anthocyanins and lycopene contents were analyzed using high-performance liquid chromatography and electrospray ionization mass spectrometry. Eleven types of anthocyanins were identified, including delphinidin, petunidin, and malvidin. Further, the antioxidant activity of anthocyanins was evaluated using 2,2′-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt radical quenching assays and electron spin resonance. “Blue tomato” extracts exert antioxidant activity. Thus, we showed that petunidin was present in the “blue tomato” peel while lycopene was present in the peel and pulp. Additionally, the blue tomato peel extract was found to significantly inhibit H₂O₂-induced cell death in vitro. This is the first study on cell protective effects of Japanese blue tomato extract and petunidin in murine photoreceptor cells.     

Attraction of the West Indian fruit fly to mango fruit volatiles

In this study, we investigated the attraction of West Indian fruit fly, Anastrepha obliqua (Macquart) (Diptera: Tephritidae), to volatiles of three mango [Mangifera indica L. (Anacardiaceae)] cultivars in field cage tests. The number of flies captured with Multilure traps baited with Amate mature green mangoes was significantly higher than that captured in traps baited with Coche and Ataulfo fruits. There was no significant difference between the number of flies captured in traps baited with Coche or Ataulfo mangoes. Gas chromatography‐mass spectrometry analysis of mango fruit volatiles identified 24, 22, and 19 compounds for Amate, Ataulfo, and Coche mango cultivars, respectively. A principal component analysis of the volatiles revealed that the Amate mango was more distant from the Ataulfo mango, and the latter cultivar was closer to the Coche mango. The compounds myrcene, α‐pinene, β‐selinene, and trans‐β‐ocimene were the most abundant in Amate mangoes, whereas 3‐carene, β‐selinene, terpinolene, and α‐pinene were the predominant compounds of Ataulfo cultivars. In the Coche mango, the predominant compounds were 3‐carene, β‐selinene, terpinolene, and limonene. Traps baited with a blend of myrcene, α‐pinene, and trans‐β‐ocimene captured more A. obliqua females and males than control traps. Flies were more attracted to the Super Q volatile extracts of Amate mango than to the three‐component blend formulated in a ratio of 1:1:1. However, there was no significant difference between the number of flies caught by traps baited with Amate mango extracts and that caught by traps baited with the three‐blend component when this was formulated according to the relative proportions in the mango extracts. Traps baited with myrcene, the major component, caught fewer flies than traps baited with Amate mango extracts.     

黄瓜化学成分的提取与研究

本文分别采用减压蒸馏-冷冻浓缩、微波-超声波萃取和固相微萃取(SPME)技术,对鲜黄瓜(带皮)、黄瓜皮、肉、籽进行了化学成分的提取及气相色谱-质谱(GC-MS)分析,共检测出109种化合物,其中大部分化合物尚未从黄瓜中分离得到.实验结果证明黄瓜皮及黄瓜籽中含有较多芳香类物质、生育酚、角鲨烯、姜烯、阿茶碱、亚油酸与油酸等营养物质.     

SENSORY EVALUATION OF MANGO DRINKS USING FUZZY LOGIC

Three brands of ready to serve mango drinks namely 'Frooti', 'Real'and 'Slice'that are available in the market were used for sensory evaluation along with the vacuum dried reconstituted mango powder drink. Mango powder was produced from mango pulp of 'Totapuri'variety with addition of Maltodextrin, Glycerol monostearate and Tri calcium phosphate at rate of 0.62, 0.015 and 0.015 kg per kg dry mango solid, respectively. These ingredients were added for getting a nonsticky free flowing powder. The pulp along with the ingredients was dried in a vacuum drier at 70 ± 2C and 710–750 mm Hg vacuum. Fuzzy logic analysis was used for finding out the best of the three market mango drinks. BrimA index (a criterion for acceptance of fruit juice) and total solid (kg per kg drink) of the reconstituted mango drink was adjusted based on the BrimA index and total solid values of the best market drink. The reconstituted mango drink satisfied the quality criteria set for 'mouth feel'but not the color, smell and taste.     

Arsenic in orchard and potato soils and plant tissue

Summary The total and water soluble arsenic contents of orchard and potato field soils are significantly higher where arsenic has been applied. Orchards with a record of past and present applications show average values as high as 99.57 ppm total arsenic and 1.99 ppm water-soluble arsenic. Orchards with a low or no arsenic application show 27.21 ppm total and 0.20 ppm water-soluble arsenic. Potato fields situated on ex-orchard plantings have an average of 38.70 ppm total and 0.32 ppm water-soluble arsenic, whereas those on non-orchard soils showed values of 9.63 and 0.04 ppm respectively. Highest soil arsenic concentrations were found in the 0–10 cm surface layer and at a distance of one meter from the tree trunk. In plant tissue the apple leaf, peel and pulp all were significantly higher for recently arsenic treated orchards, while only the apple pulp showed significance where applications had been discontinued for the past 10–15 years. Potato leaf and peel were significantly higher in arsenic on exorchard soil while the pulp was not.     

Phenolic compounds in peach (Prunus persica) cultivars at harvest and during fruit maturation

Six peach and six nectarine cultivars were evaluated for the phenolic content in their pulp and peel tissues. Chlorogenic acid, catechin, epicatechin, rutin and cyanidin-3-glucoside were detected as the main phenolic compounds of ripened fruits. The concentration was always higher in peel tissue, with average values ranging from 1 to 8 mg g⁻¹ dry weight (DW) depending on cultivar. Of the tested varieties, the white-flesh nectarine 'Silver Rome' emerged as the cultivar with the highest amount of total phenolics. Phenolic compounds were also profiled during fruit growth and ripening in the yellow nectarine cv. 'Stark Red Gold', which showed a decreasing concentration during fruit development in both peel and pulp tissues. Average amounts of total phenolics were approximately 25 mg g⁻¹ DW 60 days after full bloom and decreased to 3 mg g⁻¹ DW at ripening in pulp tissue. Differences among peel and pulp composition show the different dietetic and antioxidant potential of fruits consumed unpeeled and peeled.