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At least 19 recordsLinked to original sources
PubMed2004

Absorption, excretion, and distribution of dietary antioxidant betalains in LDLs: potential health effects of betalains in humans.

BACKGROUND: Betalains were recently identified as natural antioxidants. However, little is known about their bioavailability from dietary sources. OBJECTIVE: The objective was to evaluate the bioavailability of betalains from dietary sources. DESIGN: The plasma kinetics and urinary excretion of betalains were studied in healthy volunteers (n = 8) after a single ingestion of 500 g cactus pear fruit pulp, which provided 28 and 16 mg indicaxanthin and betanin, respectively. The incorporation of betalains in LDL and the resistance of the particles to ex vivo-induced oxidation was also researched. RESULTS: Betanin and indicaxanthin reached their maximum plasma concentrations 3 h after the fruit meal and declined according to first-order kinetics. The half-life of betanin (0.94 +/- 0.07 h) was shorter than that of indicaxanthin (2.36 +/- 0.17 h). Both compounds had disappeared from plasma by 12 h after intake. The urinary excretion of indicaxanthin and betanin over 12 h represented 76 +/- 3.0% and 3.7 +/- 0.2%, respectively, of the ingested compounds. LDL isolated 3 and 5 h after the fruit meal incorporated betalains at concentrations of 100.5 +/- 11 and 50 +/- 7.2 pmol/mg LDL protein, respectively. In addition, the particles appeared more resistant to ex vivo-induced oxidative injury than did the samples isolated before fruit ingestion (P < 0.05)-the higher the amount of betalains incorporated, the higher the resistance. The concentrations of vitamin E and beta-carotene in LDL did not change significantly after fruit ingestion. CONCLUSION: Our results show that cactus pear fruit is a source of bioavailable betalains and suggest that indicaxanthin and betanin may be involved in the observed protection of LDL against ex vivo-induced oxidative modifications.

Adult↗
PubMed2003

Recent advances in betalain research.

Betalains replace the anthocyanins in flowers and fruits of plants of most families of the Caryophyllales. Unexpectedly, they were also found in some higher fungi. Whereas the anthocyanin-analogous functions of betalains in flower and fruit colouration are obvious, their role in fungi remains obscure. The nature of newly identified betalains as well as final structure elucidation of earlier putatively described compounds published within the last decade is compiled in this report. Recent advances in research on betalain biosynthesis is also covered, including description of some 'early' reactions, i.e. betalain-specific dopa formation in plants and fungi and extradiolic dopa cleavage in fungi. Work on betalain-specific glucosyltransferases (GTs) has given new insights into the evolution of secondary plant enzymes. It is proposed that these GTs are phylogenetically related to flavonoid GTs. It was found that the decisive steps in betalain biosynthesis, i.e. condensation of the betalain chromophore betalamic acid with cyclo-dopa and amino acids or amines in the respective aldimine formation of the red-violet betacyanins and the yellow betaxanthins, are most likely to be non-enzymatic. Betalains have attracted workers in applied fields because of their use for food colouring and their antioxidant and radical scavenging properties for protection against certain oxidative stress-related disorders.

Acyltransferases↗
PubMed2005

Urinary pharmacokinetics of betalains following consumption of red beet juice in healthy humans.

The aim of the present pilot study was to characterise the renal elimination of betalains after consumption of red beet juice (RBJ). Six healthy, non-smoking female volunteers were given a single oral dose of either 500 mL of a commercial RBJ containing 362.7 mg of betalains and 500 mL of tap water, respectively, in a sequential manner. Urine was collected in intervals up to 24 h post-dose. Renal excretion of betalains was determined spectrophotometrically and quantified as betanin-equivalents. In addition, the identity of individual compounds was confirmed by HPLC coupled with diode-array detection and positive ion electrospray mass spectrometry, respectively. The amount (mean+/-S.D.) of intact betalains (betanin and isobetanin) recovered in urine was 1001+/-273 microg corresponding to 0.28+/-0.08% of the administered dose. Maximum excretion rates were observed after a median tmax,R of 3.0 h (range 2.5-8.0 h) amounting to 91.7+/-30.1 microg/h. The terminal elimination rate constant (lambdaz) and the corresponding half-life were 0.097+/-0.021 h(-1) and 7.43+/-1.47 h, respectively. Using the lambdaz estimates obtained the expected total betalain amount excreted in urine was 1228+/-291 microg. Based on the results obtained it is assumed that either the bioavailability of the betalains is low or that renal clearance is a minor route of systemic elimination for these compounds. The urinary excretion rates of unmetabolised betalains were fast and appeared to be monoexponential suggesting a one-compartment model. In order to get a more complete picture of the pharmacokinetics and health-promoting properties of red beet betalains, quantitative data on betalain bioavailability should include measurements of unchanged compounds and their corresponding metabolites in plasma, urine and bile.

Adult↗
PubMedDate not supplied

In situ and Ex situ adsorption and recovery of betalains from hairy root cultures of Beta vulgaris.

Various adsorbents were screened for in situ recovery of betalain pigments effluxed from hairy root cultures of red beet, Beta vulgaris. Alumina/silica (1:1) appeared ideal, showing in situ adsorption of 97% in a unit time of 30 min accounting for in situ recovery of 71.39% of the total betalaine effluxed. Other adsorbents such as Amberlite series (XAD-2 and -4), cyclodextrin, maltodextrin, dextrin white, and starches such as wheat starch and corn starch exhibited very poor in situ adsorption properties. Pretreatment of adsorbents with methanol significantly improved the adsorption capacities of some of the adsorbents, with a highest adsorption of 97.2% for alumina followed by alumina/silica (1:1) and higher adsorption by XAD-2 and -4. Complete in situ adsorption equilibrium was reached in 20 min for a solution containing 2.5 mg mL(-)(1) of betalain in adsorbents alumina, silica, and a mixture of alumina and silica. In situ betalain adsorption parameters for alumina/silica were determined using the Langmuir isotherm model where the adsorption capacity was found to be 0.174 mg g(-)(1) and the adsorption energy was 0.9 at pH 5.5 and 25 degrees C. Desorption of pigments from the adsorbents was invariably highest in poor adsorbents, indicating their poor adsorption energy for betalaines. Similarly, recovery by desorption was low in those adsorbents having high adsorption capacity, indicating that adsorbents such as activated ones with highest adsorption capacity with zero desorption property were unsuitable for the recovery of effluxed pigments. Ex situ recovery of betalain done using various combinations of alumina/silica and processed sand and different column geometries indicated that alumina with processed sand at a 2:1 ratio (w/w) and a minimum column material of 2 cm height and 2 cm diameter was good enough to cause 97% pigment adsorption from a solution containing 1.6 mg mL(-)(1). Desorption and recovery of pigments ex situ from columns were affected by various elution mixtures, where a gradient elution with ascending levels of HCl/ethanol in water resulted in 100% recovery of adsorbed pigments in a significantly lesser volume of eluent in a short period of 1 h. Different pigment flow rates of 0.2, 0.3, and 3.1 mL s(-)(1) through a column of alumina/processed sand indicated that a pigment equilibrium concentration of 0.18 mg mL(-)(1) at flow rates of 0.02 and 0.3 mL s(-)(1) resulted in a breakthrough at 110 and 14 min adsorbing 16.9 and 16.91 mg g(-)(1) betalain, respectively. From the breakthrough curves, the column capacities for respective flow rates were calculated as 8.86 and 9.6 mg g(-)(1), and the higher flow rates resulted in earlier breakthrough with lower capacity. Observations made in the present study are useful to develop a process for the on-line recovery of betalains effluxed from hairy roots.

Adsorption↗
PubMed2005

Color, betalain pattern, and antioxidant properties of cactus pear (Opuntia spp.) clones.

Total phenolics, ascorbic acid, and betalain contents of differently colored cactus pear clones (nine Opuntia ficus-indica [L.] Mill. clones and one O. robusta Wendl. clone) were investigated and related to their respective antioxidant potential assessed by Trolox-equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays. TEAC and ORAC values were very highly correlated with each other and also with values for total phenolics, betalain contents, and ascorbic acid concentrations. Total phenolics had the greatest contribution to ORAC and TEAC values. High-performance liquid chromatography (HPLC)-diode array detector (DAD)-tandem mass spectrometry (MS/MS) measurements of cactus pear juices permitted the differentiation of the clones based on variations in pigment patterns and betalain concentrations. The red and yellow betalains were absent in lime green colored cactus fruits. The ratio and concentration of these pigments were responsible for the yellow, orange, red, and purple colors in the other clones. Progeny of purple and lime green colored parents were characterized by 12% and 88% of plants bearing lime green and purple fruit, respectively. This implies that the genes for betalain production were lacking in the lime green fruits but could be provided by a parent with a complete set of genes, that is, purple fruits. Besides known pigments typical of Cactaceae, two unexpected betalains were identified. Whereas gomphrenin I was found for the first time in tissues of cactus plants, methionine-betaxanthin has never been described before as a genuine betalain. In addition to their alleged health-promoting properties, various combinations of yellow betaxanthins and red-purple betacyanins may allow the development of new food products without using artificial colorants.

Antioxidants↗
PubMed2003

Antioxidant activity of betalains from plants of the amaranthaceae.

Antioxidant activity of betalain pigments (seven pure compounds and four combined fractions) from plants of the family Amaranthaceae was evaluated using the modified DPPH(*) (1,1-diphenyl-2-picrylhydrazyl) method. All tested betalains exhibited strong antioxidant activity. Their EC(50) values ranged from 3.4 to 8.4 microM. Gomphrenin type betacyanins (mean = 3.7 microM) and betaxanthins (mean = 4.2 microM) demonstrated the strongest antioxidant activity, 3-4-fold stronger than ascorbic acid (13.9 microM) and also stronger than rutin (6.1 microM) and catechin (7.2 microM). Antioxidant activity of the tested betalains decreased in the following order: simple gomphrenins > acylated gomphrenins > dopamine-betaxanthin > (S)-tryptophan-betaxanthin > 3-methoxytyramine-betaxanthin > betanin/isobetanin > celosianins > iresinins > amaranthine/isoamaranthine. This study also investigated and discussed the relationship between the chemical structure and the activity of the betalains. The free radical scavenging activity of the betalains usually increased with the numbers of hydroxyl/imino groups and, moreover, depended on the position of hydroxyl groups and glycosylation of aglycones in the betalain molecules.

Amaranthaceae↗
PubMed2005

Distribution of betalain pigments in red blood cells after consumption of cactus pear fruits and increased resistance of the cells to ex vivo induced oxidative hemolysis in humans.

Betalain pigments are bioavailable phytochemicals recently acknowledged as natural radical scavengers. This work, which extends previous research on the postabsorbitive fate of dietary betalains, investigated the distribution of betanin and indicaxanthin in red blood cells (RBCs) isolated from healthy volunteers (n = 8), before and during the 1-8 h interval after a cactus pear fruit meal, and the potential antioxidative activity of the pigments in these cells. A peak concentration of indicaxanthin (1.03 +/- 0.2 microM) was observed in RBCs isolated at 3 h after fruit feeding, whereas the concentration at 5 h was about half, and even smaller amounts were measured at 8 h. Indicaxanthin was not detected at 1 h. Betanin (30.0 +/- 5.2 nM) was found only in RBCs isolated at 3 h from fruit feeding. In comparison with homologous RBCs before fruit ingestion, a significant delay (P < 0.05) of the onset of an ex vivo cumene hydroperoxide (cumOOH)-induced hemolysis was evident in the RBCs isolated at 3 h (33.0 +/- 4.5 min) and at 5 h (16.0 +/- 2.0 min). Neither vitamins C and E nor GSH was modified in the RBCs at any time point. Blood collected from the same volunteers after a 12-h fasting was incubated with the purified betalains in the range of 5-25 microM, to enrich the erythrocytes with either betanin or indicaxanthin, and then the cells were exposed to cumOOH. When compared to the relevant nonenriched cells, the betalain-enriched erythrocytes exhibited an enhanced resistance to the cumOOH-induced hemolysis, which was positively correlated (r (2) = 0.99) to the amount of the incorporated compound. On a micromolar basis, betanin and indicaxanthin showed a comparable effectiveness. Taken together, these findings provide evidence that human RBCs incorporate dietary betalains and support the concept that these phytochemicals may offer antioxidative protection to the cells.

Adult↗
PubMed2004

Characterization and functional identification of a novel plant 4,5-extradiol dioxygenase involved in betalain pigment biosynthesis in Portulaca grandiflora.

Betalains are pigments that replace anthocyanins in the majority of families of the plant order Caryophyllales. Betalamic acid is the common chromophore of betalains. The key enzyme of the betalain biosynthetic pathway is an extradiol dioxygenase that opens the cyclic ring of dihydroxy-phenylalanine (DOPA) between carbons 4 and 5, thus producing an unstable seco-DOPA that rearranges nonenzymatically to betalamic acid. A gene for a 4,5-DOPA-dioxygenase has already been isolated from the fungus Amanita muscaria, but no homolog was ever found in plants. To identify the plant gene, we constructed subtractive libraries between different colored phenotypes of isogenic lines of Portulaca grandiflora (Portulacaceae) and between different stages of flower bud formation. Using in silico analysis of differentially expressed cDNAs, we identified a candidate showing strong homology at the level of translated protein with the LigB domain present in several bacterial extradiol 4,5-dioxygenases. The gene was expressed only in colored flower petals. The function of this gene in the betalain biosynthetic pathway was confirmed by biolistic genetic complementation in white petals of P. grandiflora genotypes lacking the gene for color formation. This gene named DODA is the first characterized member of a novel family of plant dioxygenases phylogenetically distinct from Amanita sp. DOPA-dioxygenase. Homologs of DODA are present not only in betalain-producing plants but also, albeit with some changes near the catalytic site, in other angiosperms and in the bryophyte Physcomitrella patens. These homologs are part of a novel conserved plant gene family probably involved in aromatic compound metabolism.

Amino Acid Sequence↗
PubMed2002

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↗
PubMedDate not supplied

Food-grade chemical and biological agents permeabilize red beet hairy roots, assisting the release of betalaines.

Hairy root cultures of red beet, Beta vulgaris L., were permeabilized under the functions of food-grade chemical and biological agents cetyl trimethylammonium bromide (CTAB), Triton X-100, Tween-80, Lactobacillus helveticus, Saccharomyces cereviseae, and Candida utilis, as well as cell fractions of L. helveticus, for the recovery of betalaines with or without oxygen stress. Tween-80 (0.15%), Triton X-100 (0.2%), and CTAB (0.05%), in combination with oxygen stress, released 45%, 70%, and 90% pigment into the medium, respectively, with significantly lesser levels in agitated cultures receiving similar treatments. The release was rapid (1 h) in CTAB treatment with a much slower release in Tween-80. CTAB (0.002%) was found to be also useful in effluxing betalaines (80%) from hairy roots grown in a bubble column reactor. Viability of permeabilized hairy roots, tested on agar medium, was not affected by any level of CTAB treatment and was significantly retarded at higher levels of Triton X-100 and Tween-80. An altogether new approach of pigment release using biological agents such as live cells of food-grade microbes was used where C. utilis, L. helveticus, and S. cereviseae released 60%, 85%, and 54% betalaines, respectively, in 24 h, though lower level treatments also released similar levels of pigment by 48 h. Dried whole cell powder of L. helveticus, its total insoluble carbohydrate, and free lipid fractions released 10%, 0%, and 85% pigment, respectively. An extended study with a bubble column reactor using the free lipid fraction of L. helveticus showed 50% and 84% pigment release in 8 and 12 h, respectively, exhibiting good viability when plated on agar medium. Even in the bioreactor, replenishment of medium 8 h after treatment with free lipid of L. helveticus allowed regrowth of hairy roots. The high level of pigment release recorded here, using CTAB or lipid of L. helveticus, appears useful for developing processes for in situ recovery of betalaines. The live microbes, applicable only for batch cultures, are expected to impart improved sensory/nutraceutical effects to the recovered pigment and hence may add value to the product receiving the red beet pigment thus produced.

Beta vulgaris↗
PubMed2003

Increased resistance to oxidation of betalain-enriched human low density lipoproteins.

Betalains are natural pigments recently considered as compounds with potential antioxidative properties. In this work, ex vivo plasma spiking of pure either betanin or indicaxanthin, followed by isolation of low density lipoprotein (LDL), and measurement of its resistance to copper-induced oxidation, has been used to research if these betalains can bind to LDL and prevent oxidation of LDL lipids. When pooled human plasma from 10 healthy volunteers was incubated in the presence of 25-100 microM either betanin or indicaxanthin, incorporation of both compounds in LDL was observed, with a maximum binding of 0.52 +/- 0.08, and 0.51 +/- 0.06 nmoles of indicaxanthin and betanin, respectively, per mg LDL protein. Indicaxanthin-enriched and betanin-enriched LDL were more resistant than homologous native LDL to copper-induced oxidation, as assessed by the elongation of the induction period. The incorporated indicaxanthin, however, appeared twice as effective as betanin in increasing the length of the lag phase, while both compounds did not affect the propagation rate. Both betalains were consumed during the inhibition period of lipid oxidation, and delayed consumption of LDL-beta carotene. Indicaxanthin, but not betanin, prevented vitamin E consumption at the beginning of LDL oxidation, and prolonged the time of its utilization. The resistance of LDL to oxidation when vitamin E and indicaxanthin acted separately in a sequence, was lower than that measured when they were allowed to act in combination, indicating some synergistic interaction between the two molecules. No prooxidant effect over a large concentration range of either betanin or indicaxanthin was observed, when either betalain was added to the LDL system undergoing a copper-induced oxidation. These results show than indicaxanthin and betanin may bind to LDL, and are highly effective in preventing copper-induced lipid oxidation. Interaction with vitamin E appears to add a remarkable potential to indicaxanthin in the protection of LDL. Although molecular mechanisms remain uncompletely understood, various aspects of the action of betanin and indicaxanthin in preventing LDL lipid oxidation are discussed.

Antioxidants↗
PubMed2005

Radical scavenging activity and stability of betalains from Beta vulgaris hairy root culture in simulated conditions of human gastrointestinal tract.

Stability and radical scavenging activity of betalains, biosynthesized by Beta vulgaris hairy root culture were investigated at different pH values, bile salts concentration, as well as at the in vitro conditions of gastrointestinal tract. It was established that pH below 3 and concentrations of the bile salts up to 4% had no great influence on betalains stability. At the in vitro conditions of gastrointestinal tract betalains are relatively stable, as their radical scavenging activity decrease from 75% inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH*) to about 38%.

Beta vulgaris↗
PubMed2002

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↗
PubMed2004

Identification of betalains from petioles of differently colored Swiss chard (Beta vulgaris L. ssp. cicla [L.] Alef. Cv. Bright Lights) by high-performance liquid chromatography-electrospray ionization mass spectrometry.

The betalain pattern of differently colored Swiss chard (Beta vulgaris L. ssp. cicla [L.] Alef. cv. Bright Lights) was investigated for the first time. Nineteen betaxanthins and nine betacyanins were identified by RP-HPLC and positive ion electrospray mass spectrometry, co-injection experiments with semisynthetic reference compounds, and standards derived from authentic plant material, respectively. Histamine-betaxanthin and alanine-betaxanthin were found to be novel betaxanthins, which to the best of our knowledge have not been reported as natural compounds until now. Furthermore, tyramine-betaxanthin (miraxanthin III) and 3-methoxytyramine-betaxanthin, which to date were known only from families other than the Chenopodiaceae, were detected for the first time in colored Swiss chard. The betacyanin pattern of purple petioles was composed of betanin, isobetanin, betanidin, and isobetanidin. Although phyllocactin was present in only trace amounts, further acylated structures such as betanidin-monoferuloyl-5-O-beta-diglucoside and lampranthin II, accompanied by their corresponding C(15)-epimers, were identified. In addition, quantification of betalains and CIE LCh degrees measurements were performed with the colored extracts to correlate the visual appearance with the respective pigment patterns. Besides the novel phytochemical findings, the present study is useful for the evaluation of betalainic Swiss chard as a potential coloring foodstuff.

Beta vulgaris↗
PubMed2005

Betaxanthins as substrates for tyrosinase. An approach to the role of tyrosinase in the biosynthetic pathway of betalains.

Tyrosinase or polyphenol oxidase (EC 1.14.18.1) is the key enzyme in melanin biosynthesis and in the enzymatic browning of fruits and vegetables. The role of tyrosinase in the secondary metabolism of plants still remains unclear, but its implication in betalain biosynthesis has been proposed. Betalains are an important class of water-soluble pigments, characteristic of plants belonging to the order Caryophyllales. In this article, the betaxanthins, tyrosine-betaxanthin (portulacaxanthin II) and dopaxanthin, are reported to be physiological substrates for tyrosinase. The direct activity of tyrosinase on selected betaxanthins is characterized in depth, and conversion of tyrosine-betaxanthin to dopaxanthin and its further oxidation to a series of compounds are described. Identity of the reaction products was studied by high-performance liquid chromatography and electrospray ionization-mass spectrometry. Masses determined for the reaction products were the same in all cases, 389 m/z ([M + H]+) and equal to that determined for betanidin. Data indicate that dopaxanthin-quinone is obtained and evolves to more stable species by intramolecular cyclization. Kinetic parameters for tyrosinase acting on dopaxanthin were evaluated, showing a high affinity for this substrate (K(m) = 84.3 microM). The biosynthetic scheme of betalains is reviewed and a branch is proposed based on the description of physiological substrates for tyrosinase. Lampranthus productus, Glottiphylum oligocarpum, and Glottiphylum pigmaeum are described as sources of stereopure (2S/S)-dopaxanthin.

Betalains↗
PubMed1997

The gene coding for the DOPA dioxygenase involved in betalain biosynthesis in Amanita muscaria and its regulation.

Genomic and cDNA clones derived from the gene (dodA) coding for DOPA dioxygenase, a key enzyme in the betalain pathway, were obtained from the basidiomycete Amanita muscaria. A cDNA library was established in the phage lambda ZapII and dodA clones were isolated using polyclonal antibodies raised against the purified enzyme. Their identity was confirmed by comparison of the deduced amino acid sequence with the sequence of several tryptic peptide fragments of DOPA dioxygenase. The gene coded for a 228-amino acid protein that showed no homology to published sequences. The coding region was interrupted by five short introns. Regulation was shown to occur at the transcriptional level; the mRNA accumulated to high levels only in the coloured cap tissue. dodA was found to be a single-copy gene in A. muscaria. To our knowledge, this is the first gene from the betalain pathway to be cloned. It encodes a type of aromatic ring-cleaving dioxygenase that has not been previously described.

Amanita↗
PubMed1983

Inability of red beet betalain pigments to initiate or promote hepatocarcinogenesis.

A short-term bioassay was used to determine the ability of red-beet betalain pigments to initiate or promote hepatocarcinogenesis in rat liver. Female Sprague-Dawley rats were partially hepatectomized and separated into nine groups (6-11 animals/group). Four of the groups were treated with betacyanin pigment preparations (betacyanin solution after fermentation, 50 mg/kg; pure betanin, 50 mg/kg; degraded betanin, 50 mg/kg; a betacyanin diet containing 2000 ppm/kg) to evaluate their ability to initiate hepatocarcinogenesis. N-Nitrosodiethylamine (NDEA: 10 mg/kg) was used for the positive control. Another group, previously initiated with NDEA, received daily a betacyanin solution (100 ppm; 3.5 mg/rat/day) to determine the pigment's ability to promote NDEA hepatocarcinogenesis in comparison with positive and negative controls treated respectively with and without a promoting agent (0.5% phenobarbital in the diet). Animals were killed after month 6 (promotion test) or 8 (initiation test). Liver sections were stained with haematoxylin and eosin and for gamma-glutamyl transpeptidase (GGT) activity. The number of enzyme-altered foci and the percentage of liver volume so affected, were determined for each group, by scoring for GGT. Comparison of the results obtained for the experimental groups with those for positive and negative control groups indicated that the betacyanin pigments tested in this assay did not initiate or promote hepatocarcinogenesis in rat liver. These findings provide further evidence that betalain colourants may be viable alternatives for synthetic dyes currently used as food additives.

Animals↗