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Cloning and expression of an acidic pectin methylesterase from jelly fig (Ficus awkeotsang).

Pectin methylesterase (PME) is the key enzyme responsible for the gelation of jelly curd in the water extract of jelly fig (Ficus awkeotasang) achenes. The jelly fig PME extracted from achenes was isoelectrofocused at pH 2.5 and subjected to N-terminal amino acid sequencing. A cDNA fragment encoding the mature protein of this acidic PME was obtained by PCR cloning using a poly(T) primer and a degenerate primer designed according to the N-terminal sequence of the purified PME. The complete cDNA sequence of its precursor protein was further obtained by PCR using the same strategy. The PME clone was overexpressed in Escherichia coli, and its expressed protein was immunologically recognized as strongly as the original antigen using antibodies against purified PME. Fractionation analysis revealed that the overexpressed PME was predominantly present in the pellet and thus presumably formed insoluble inclusion bodies in E. coli cells.

Amino Acid Sequence↗

Identification of betalains from yellow beet (Beta vulgaris L.) and cactus pear [Opuntia ficus-indica (L.) Mill.] by high-performance liquid chromatography-electrospray ionization mass spectrometry.

Betaxanthins, the yellow-orange water-soluble pigments from yellow beet (Beta vulgaris ssp. vulgaris cv. Bejo Zaden) and cactus pear (Opuntia ficus-indica cv. Gialla) have been investigated using an HPLC system compatible with mass spectrometry. Five novel betaxanthins were found and characterized as the immonium adducts of betalamic acid with serine, gamma-aminobutyric acid, valine, isoleucine, and phenylalanine. To enable concentration of betalain samples, desalting was performed by solid-phase extraction. With this technique, betacyanins could be separated from the betaxanthins using the pH-dependent retention characteristics of red and yellow betalains. The betaxanthin fraction was taken for the preparation of betalamic acid as a precursor for semisynthetic standards. The HPLC method was applied to yellow beet and cactus pear, revealing a more complex betalain profile than described earlier, thus proving its suitability for screening of betaxanthin-containing plants as potential sources for natural food colors.

Beta vulgaris↗

Thiabendazole uptake and storage performance of cactus pear [Opuntia ficus-indica (L.) Mill. Cv Gialla] fruit following postharvest treatments with reduced doses of fungicide at 52 degrees C.

The storage response of cactus pears [Opuntia ficus-indica Miller (L.) cv. Gialla] was investigated over 6 weeks at 6 degrees C, plus an additional week of simulated marketing period (SMP) at 20 degrees C, after a 3-min dip treatment with thiabendazole (TBZ) at 1000 mg/L at 20 degrees C or 150 mg/L TBZ at 52 degrees C. Untreated fruits were used as control. Following TBZ treatments at 20 and 52 degrees C, total residues were recovered from the peel of cactus pear, as the concentration of residues in the pulp was negligible. Treatments with 1000 mg/L TBZ at 20 degrees C resulted in a 2.82 mg/kg residue uptake (active ingredient, whole-fruit basis), whereas treatment at 150 mg/L TBZ left 1.09 mg/kg. TBZ showed great persistence over both storage and SMP: on average, in the fruits treated at 20 and 52 degrees C, over 72 and 68%, respectively, of TBZ was still present after SMP. Postharvest treatments with 1000 mg/L TBZ at room temperature did not affect the expression of slight-to-moderate chilling injury (CI), but reduced severe CI by approximately 50% and decay development by 63.4% in comparison to those of untreated fruit after SMP. The effectiveness of TBZ was much higher with the treatment at 150 mg/L TBZ at 52 degrees C, providing 91% control of severe CI and approximately 89% suppression of decay; no treatment damage occurred during storage and SMP. External appearance was better in fruit treated with 150 mg/L TBZ at 52 degrees C. Respiration rate, titratable acidity, soluble solids concentration, and acetaldehyde in the flesh were not significantly influenced by treatments. Ethylene production rate and ethanol levels in the flesh were significantly higher in the TBZ-treated fruit as opposed to those in the untreated control fruit.

Cold Temperature↗

Antioxidant property of an ethanol extract of the stem of Opuntia ficus-indica var. saboten.

An ethanol extract of the stem of Opuntia ficus-indica var. saboten (OFS) was assessed to determine the mechanism(s) of its antioxidant activity. The ethanol extract exhibited a concentration-dependent inhibition of linoleic acid oxidation in a thiocyanate assay system. In addition, the OFS extract showed dose-dependent free-radical scavenging activity, including DPPH radicals, superoxide anions (O(2)(*-)), and hydroxyl radicals (*OH), using different assay systems. The OFS ethanol extract was also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals in a Fenton's reaction mixture. Furthermore, the extract showed significant (p < 0.01) dose-dependent protection of mouse splenocytes against glucose oxidase-mediated cytotoxicity. Finally, the OFS extract was characterized as containing a high amount of phenolics (180.3 mg/g), which might be the active compounds responsible for the antioxidant properties of the OFS extract.

Animals↗

Antioxidant activities of sicilian prickly pear (Opuntia ficus indica) fruit extracts and reducing properties of its betalains: betanin and indicaxanthin.

Sicilian cultivars of prickly pear (Opuntia ficus indica) produce yellow, red, and white fruits, due to the combination of two betalain pigments, the purple-red betanin and the yellow-orange indicaxanthin. The betalain distribution in the three cultivars and the antioxidant activities of methanolic extracts from edible pulp were investigated. In addition, the reducing capacity of purified betanin and indicaxanthin was measured. According to a spectrophotometric analysis, the yellow cultivar exhibited the highest amount of betalains, followed by the red and white ones. Indicaxanthin accounted for about 99% of betalains in the white fruit, while the ratio of betanin to indicaxanthin varied from 1:8 (w:w) in the yellow fruit to 2:1 (w:w) in the red one. Polyphenol pigments were negligible components only in the red fruit. When measured as 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) equivalents per gram of pulp, the methanolic fruit extracts showed a marked antioxidant activity. Vitamin C did not account for more than 40% of the measured activity. In addition, the extracts dose-dependently inhibited the organic hydroperoxide-stimulated red cell membrane lipid oxidation, as well as the metal-dependent and -independent low-density lipoprotein oxidation. The extract from the white fruit showed the highest protection in all models of lipid oxidation. Purified betanin and indicaxanthin were more effective than Trolox at scavenging the [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] diammonium salt cation radical. Cyclic voltammetric measurements show two anodic waves for betanin and indicaxanthin, and differential pulse voltammetry shows three anodic waves for betanin, with calculated peak potentials of 404, 616, and 998 mV, and two anodic waves for indicaxanthin, with peak potentials of 611 and 895 mV. Betanin underwent complex formation through chelation with Cu(2+), whereas indicaxanthin was not modified. These findings suggest that the above betalains contribute to the antioxidant activity of prickly pear fruits.

Antioxidants↗

Chemical characterization and biological effects of Sicilian Opuntia ficus indica (L.) mill. Fruit juice: antioxidant and antiulcerogenic activity.

The juice of whole fruits of Sicilian cultivars of prickly pear (Opuntia ficus indica (L.) Mill.) was investigated, and the contents of ascorbic acid, total polyphenols, and flavonoids were determined. In the juice, ferulic acid was the chief derivative of hydroxycinnamic acid and the mean concentration of total phenolic compounds was 746 microg/mL. The flavonoid fraction, analyzed by high-performance liquid chromatography-diode array detection, consisted of rutin and isorhamnetin derivatives. The juice showed antioxidant activity in the DPPH(*) test, probably due to the phenolic compounds that are effective radical scavengers. The preventive administration of the juice inhibited the ulcerogenic activity of ethanol in rat. Light microscopy observations showed an increase in mucus production and the restoration of the normal mucosal architecture. The juice is nutritionally interesting, and its dietary intake could provide protection against oxidative damage.

Animals↗

Chemical characterization of sicilian prickly pear (Opuntia ficus indica) and perspectives for the storage of its juice.

In this work, Sicilian cultivars of prickly pear (Opuntia ficus indica) were partially characterized from a chemical point of view, and the possibility of long-term storage of their juice was investigated. The acidity of the prickly pear juice turned out to be very low (0.02%) and the pH very high (6.4-6.5) if compared with values found in other common fruit juices. In the perspective of processing and storage conditions according to Italian law, the acidity has been corrected by adding the proper amount of tartaric and/or phosphoric acid. The sugar content (mainly glucose and fructose) is very high (11-12%), and also L-ascorbic acid is present in considerable amount (31-38 mg/100 g). Among the transition metals, a high content of manganese(II) (1.7-2.9 ppm) and good amounts of iron(III) (0.6-1.2 ppm) and zinc(II) (0.3-0.4 ppm) were found. In particular, such ions appear to be present mainly in the thick skin of the fruit or "trapped" inside the pulp. Pectin methylesterase (PME) seems to be present in very small amount and/or is not highly active. Furthermore, PME activity decreases considerably after the necessary adjustment of the pH and the thermal treatment requested for long-term storage. After approximately 2 months, none of the juices prepared was affected by noticeable sedimentation of the pulp. Finally, different samples of prickly pear juice were sensorially analyzed, employing descriptors such as color, aroma, viscosity, acidity, sweetness, and off-flavors. The results obtained can be considered very satisfactory, and the juice has been widely appreciated when compared with other products commonly available on the market such as pear and peach juices.

Ascorbic Acid↗

New peroxy triterpenes from the aerial roots of Ficus microcarpa.

Six new triterpenes, 3 beta-acetoxy-12 beta,13 beta-epoxy-11 alpha-hydroperoxyursane (1), 3 beta-acetoxy-11 alpha-hydroperoxy-13 alpha H-ursan-12-one (2), 3 beta-acetoxy-1 beta,11 alpha-epidioxy-12-ursene (3), (20S)-3 beta-acetoxylupan-29-oic acid (4), (20S)-3 beta-acetoxy-20-hydroperoxy-30-norlupane (5), and 3 beta-acetoxy-18 alpha-hydroperoxy-12-oleanen-11-one (6), together with 3 beta-acetoxy-12-oleanen-11-one (7), were isolated from the aerial roots of Ficus microcarpa. Compounds 1-3, 5, and 6 were characterized as new peroxytriterpenes. The structures of 3 and 6 were confirmed by X-ray crystallography, and their structures were elucidated by spectroscopic and chemical methods.

Crystallography, X-Ray↗

Taraxastane-type triterpenes from the aerial roots of Ficus microcarpa.

Seven new taraxastane-type triterpenes-20-taraxastene-3beta, 22alpha-diol (1), 3beta-acetoxy-20-taraxasten-22alpha-ol (2), 3beta-acetoxy-22alpha-methoxy-20-taraxastene (3), 3beta-acetoxy-20alpha,21alpha-epoxytaraxastan-22alp ha-ol (4), 3beta-acetoxy-19alpha-methoxy-20-taraxastene (5), 3beta-acetoxy-19alpha-hydroperoxy-20-taraxastene (6), 3beta-acetoxy-20alpha,21alpha-epoxytaraxastane (7)-were isolated from the aerial roots of Ficus microcarpa. Their structures were elucidated by spectroscopic and chemical methods.

Magnetic Resonance Spectroscopy↗

Composition of pulp, skin and seeds of prickly pears fruit (Opuntia ficus indica sp.).

The proximate composition of pulp, skin and seeds of prickly pear cactus (Opuntia ficus indica) was investigated and is reported on a dry weight basis. The most abundant component of the pulp and skin was ethanol-soluble carbohydrates. Pulp contained glucose (35%) and fructose (29%) while the skin contained essentially glucose (21%). Protein content was 5.1% (pulp), 8.3% (skin) and 11.8% (seeds). Starch was found in each of the three parts of the fruit. Pulp fibers were rich in pectin (14.4%), skin and seeds were rich in cellulose (29.1 and 45.1%, respectively). Skin was remarkable for its content of calcium (2.09%) and potassium (3.4%). Prickly pear is a neglected nutritional source which should be more widely used because of its potential nutrient contribution.

Calcium↗

The characterization of ficaprenol-10, -11 and 12 from the leaves of Ficus elastica (decorative rubber plant).

Evidence from mass, nuclear-magnetic-resonance and infrared spectrometry and from gas-liquid and thin-layer chromatography is presented in favour of the presence of cis-trans-decaprenol, -undecaprenol and -dodecaprenol in the mixture of polyprenols (2.6mg./g.) isolated from leaf tissue of Ficus elastica. The trivial names ficaprenol-10, -11 and -12 are proposed. Nuclear-magnetic-resonance studies showed that each of these prenols contains three trans internal isoprene residues and a cis ;OH-terminal' isoprene residue. Ficaprenol-11 is the major component of the mixture. Chromatographic evidence suggests the presence also of small amounts of ficaprenol-9 and -13. The precise position of the three trans internal isoprene residues was not determined but it is suggested that these are adjacent to the omega-terminal isoprene residue and that the ficaprenols are formed from all-trans-geranylgeranyl pyrophosphate. It is also suggested that ficaprenol-10, -11, -12 and -13 are probably the same compounds as castaprenol-10, -11, -12 and -13.

Alcohols↗

Preparation of fully active ficin from Ficus glabrata by covalent chromatography and characterization of its active centre by using 2,2'-depyridyl disulphide as a reactivity probe.

1. Fully active ficin (EC 3.4.22.3) containing 1 mol of thiol with high reactivity towards 2,2'-dipyridyl disulphide (2-Py-S-S-2-Py) at pH4.5 per mol of protein was prepared from the dried latex of Ficus glabrata by covalent chromatography on a Sepharose-glutathione-2-pyridyl disulphide gel. 2. Ficin thus prepared is a mixture of ficins I-IV and ficin G, in which ficins II and III predominate. The various ficins exhibit similar reactivity characteristics towards 2-Py-S-S-2-Py. 3. Use of 2-Py-S-S-2-Py as a reactivity probe demonstrates (a) that in ficin, as in papain (EC 3.4.22.2), the active-centre thiol and imidazole groups interact to provide a nucleophilic state at pH values of approx. 6 additional to the uncomplicated thiolate ion that predominates at pH values over 9, and (b) a structural difference between ficin and papain that leads to a much higher rate of reaction of 2-Py-S-S-2-Py with ficin than with papain at pH values 3-4. This difference is suggested to include a lack in ficin of a carboxyl group conformationally equivalent to that of aspartic acid-158 in papain. 4. The high electrophilicity of the 2-Py-S-S-2PyH+ monocation allows directly the detection of the exposure of the buried thiol group of ficin at pH values below 4.

2,2'-Dipyridyl↗

A triterpene from Ficus pumila.

The leaves of Ficus pumila afforded a new neohopane (1) by silica gel chromatography. The structure of 1 was elucidated by 1D and 2D NMR and IR spectroscopy and mass spectrometry. It showed antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Candida albicans with an average antimicrobial index of 0.5, 0.3, 0.3 and 0.7, respectively, at a concentration of 30 microg.

Anti-Bacterial Agents↗

Supplementation with cactus pear (Opuntia ficus-indica) fruit decreases oxidative stress in healthy humans: a comparative study with vitamin C.

BACKGROUND: Cactus pear (Opuntia ficus-indica) fruit contains vitamin C and characteristic betalain pigments, the radical-scavenging properties and antioxidant activities of which have been shown in vitro. OBJECTIVE: We investigated the effects of short-term supplementation with cactus pear fruit compared with vitamin C alone on total-body oxidative status in healthy humans. DESIGN: In a randomized, crossover, double-treatment study, 18 healthy volunteers received either 250 g fresh fruit pulp or 75 mg vitamin C twice daily for 2 wk, with a 6-wk washout period between the treatments. Before (baseline) and after each treatment, 8-epi-prostaglandin F(2alpha) (8-epi-PGF(2alpha)) and malondialdehyde in plasma, the ratio of reduced to oxidized glutathione (GSH:GSSG) in erythrocytes, and lipid hydroperoxides in LDL were measured as biomarkers of oxidative stress; plasma Trolox-equivalent antioxidant activity (TEAC) and vitamins A, E, and C were evaluated as indexes of antioxidant status. RESULTS: Both treatments caused comparable increases compared with baseline in plasma concentrations of vitamin E and vitamin C (P < 0.05); vitamin A and TEAC did not change significantly. After supplementation with cactus pear fruit, 8-epi-PGF(2)alpha and malondialdehyde decreased by approximately 30% and 75%, respectively; GSH:GSSG shifted toward a higher value (P < 0.05); and LDL hydroperoxides were reduced by almost one-half. Supplementation with vitamin C did not significantly affect any marker of oxidative stress. CONCLUSIONS: Consumption of cactus pear fruit positively affects the body's redox balance, decreases oxidative damage to lipids, and improves antioxidant status in healthy humans. Supplementation with vitamin C at a comparable dosage enhances overall antioxidant defense but does not significantly affect body oxidative stress. Components of cactus pear fruit other than antioxidant vitamins may play a role in the observed effects.

Adult↗

Young daughter cladodes affect CO2 uptake by mother cladodes of Opuntia ficus-indica.

BACKGROUND AND AIMS: Drought damages cultivated C3, C4 and CAM plants in the semi-arid lands of central Mexico. Drought damage to Opuntia is common when mother cladodes, planted during the dry spring season, develop young daughter cladodes that behave like C3 plants, with daytime stomatal opening and water loss. In contrast, wild Opuntia are less affected because daughter cladodes do not develop on them under extreme drought conditions. The main objective of this work is to evaluate the effects of the number of daughter cladodes on gas exchange parameters of mother cladodes of Opuntia ficus-indica exposed to varying soil water contents. METHODS: Rates of net CO2 uptake, stomatal conductance, intercellular CO2 concentration, chlorophyll content and relative water content were measured in mature mother cladodes with a variable number of daughter cladodes growing in spring under dry and wet conditions. KEY RESULTS: Daily carbon gain by mother cladodes was reduced as the number of daughter cladodes increased to eight, especially during drought. This was accompanied by decreased mother cladode relative water content, suggesting movement of water from mother to daughter cladodes. CO2 assimilation was most affected in phase IV of CAM (late afternoon net CO2 uptake) by the combined effects of daughter cladodes and drought. Rainfall raised the soil water content, decreasing the effects of daughter cladodes on net CO2 uptake by mother cladodes. CONCLUSIONS: Daughter cladodes significantly hasten the effects of drought on mother cladodes by competition for the water supply and thus decrease daily carbon gain by mother cladodes, mainly by inhibiting phase IV of CAM.

Biological Transport↗

Agrobacterium larrymoorei sp. nov., a pathogen isolated from aerial tumours of Ficus benjamina.

Tumorigenic Agrobacterium strains isolated from tumours growing on pruned branches of Ficus benjamina have previously been shown to have unique opine metabolism and sufficient 16S rRNA sequence differences to suggest that they belong to a new species. DNA-DNA hybridization results confirmed that these strains represent a new species and Agrobacterium larrymoorei sp. nov. (type strain ATCC 51759T = CFBP 5473T = NCPPB 4096T) is proposed as the name for the species.

DNA, Bacterial↗

Nature of the Conversion of Ficus carica Variety Kadota Ficin Component D to Component C. Some Physicochemical Properties of Components C and D.

Component C can be formed from component D under the experimental conditions used during purification of Ficus carica variety Kadota latex. By use of the inhibitor, sodium p-chloromercuribenzoate, the 2 components have been purified to chromatographic homogeneity. The 2 components have identical molecular weights and amino acid composition. The only difference found between the 2 components is the presence of 3 to 6 more amide groups in component D than in component C. There also appears to be a conformational difference between the 2 since component C is not as acidic, with respect to component D, as would be expected from the comparative amide contents. Conformational differences between the 2 are also indicated by the chromatographic behavior of the 2 in the presence and absence of sodium-p-chloromercuribenzoate.

Journal Article↗

Growth and Respiratory Response of Fig (Ficus carica L. cv. Mission) Fruits to Ethylene.

Growth in diameter of the fig (Ficus carica L. cv. Mission) fruit takes place in three distinct periods; two periods (I and III) of rapid growth are separated by a period (II) of slow growth. With respect to exposure to ethylene, the fruit exhibits a two phase response. Ethylene inhibits fruit growth in phase A (period I), the period of cell division, stimulates growth in early phase B (early period II), and stimulates both growth and ripening during the remainder of phase B (late period II and period III). The adverse effect of exogenous ethylene on fruits during phase A is thought to be due to inhibition of cell division. The gradual transition occurring in the response of fruits during phase B was interpreted in terms of carbohydrate level in the fruits.The onset of period III and a respiratory climacteric rise was preceded by or concomitant with a sudden burst of endogenous ethylene synthesis. This, together with the fact that exogenous ethylene applied at the proper stage of fruit growth triggers both ripening and the climacteric rise, leads to the conclusion that ethylene is the causal agent. In other words, the data support the concept that ethylene is a growth hormone that initiates a chain of metabolic and physiological events leading to fig fruit ripening.

Journal Article↗