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At least 19 recordsLinked to original sources
PubMed2000

Tissue and intracellular localization of indican and the purification and characterization of indican synthase from indigo plants.

Indican (indoxyl beta-D-glucoside) was found to accumulate only in green leaves of the indigo plant, and not in any other tissues. Comparisons of the indican content of protoplasts and vacuoles showed that indican was stored only in the vacuole of the cell. Indican content appeared and increased with the appearance and growth of leaves. In mature plants, the younger leaves contained larger amounts of indican than the older ones. Cell extracts of young leaves of indigo plant catalyzed the synthesis of indican from UDP-glucose and indoxyl. Indican synthase was extracted and purified from young leaves. The enzyme was separated into two fractions by anion-exchange chromatography. The enzyme in the fraction which was eluted by 0.1 M NaCl had a molecular weight of 53,000 by SDS-PAGE. Optimum pH of the enzyme was at about 10.0, indicating that the enzyme is likely localized in a different intracellular compartment from that of indican storage. The enzyme showed normal Michaelis-Menten kinetics and a K(m) value of 0.13 mM for UDP-glucose.

Glucosyltransferases↗
PubMedDate not supplied

A new HPLC-ELSD method to quantify indican in Polygonum tinctorium L. and to evaluate beta-glucosidase hydrolysis of indican for indigo production.

A method to quantify the indigo precursor indican (indoxyl-beta-D-glucoside) in Polygonum tinctorium L. has been developed. Plant material was extracted in deionized water, and indican was identified and quantified using high performance liquid chromatography (HPLC) coupled to an evaporative light scattering detector (ELSD). Results confirmed that with this method it is possible to measure indican content in a short time, obtaining reliable and reproducible data. Using this method, leaf indican content was quantified every 15 days during the growing season (from May to October) in P. tinctorium crops grown in a field experiment in Central Italy. Results showed that indican increased along the growing season until flowering and was positively affected by photosynthetic active radiation (PAR). Indican is naturally hydrolyzed by native beta-glucosidase to indoxyl and glucose, the indoxyl yielding indigo. The activity of two enzymes, sweet almond beta-glucosidase and Novarom G preparation, were compared with P. tinctorium native beta-glucosidase to evaluate indigo production. Results showed that the ability to promote indigo formation increased as follows: almond beta-glucosidase <or= Novarom G.

Chromatography, High Pressure Liquid↗
PubMed1981

Effect of lactobacilli on urinary indican excretion in gnotobiotic rats and in man.

The effect of Lactobacillus species on urinary indican excretion in two types of gnotobiotic rats (GBH-9 and GB-5) and in man was studied. L. salivarius, L. plantarum and L. casei colonized in the cecum and colon of both types of rats at levels of 10(7) to 10(8) per gram of contents following one-dose oral administration and caused a significant reduction in urinary indican excretion, whereas two strains of L. acidophilus which colonized at low levels (10(4) to 10(5)/g) did not reduce indican excretion. Daily feeding of Lactobacillus concentrates caused a further significant reduction in indican excretion which was corrected by tryptophan intake, even in the case of L. acidophilus. There was a clear relationship between fecal lactobacillus counts and urinary indican. A significant reduction of indican secretion was obtained only when the fecal counts of lactobacilli exceeded 10(7).4/g in rats. L. casei also had an effect in man, reducing urinary excretion of indican and p-cresol. Reduction in indican excretion seems to be accompanied by decreasing fecal tryptophanase activity in rats and man. A negative correlation was also obtained between fecal resident lactobacilli and urinary indican in man (r = -0.532, n = 28, p less than 0.01). Subjects with lactobacilli at a level of 10(6)/g or more excreted less indican than those harboring the bacteria at levels below 10(5)/g.

Adult↗
PubMed2004

Environmental factors affecting productivity, indican content, and indigo yield in Polygonum tinctorium Ait., a subtropical crop grown under temperate conditions.

Polygonum tinctorium Ait. is a herbaceous subtropical annual plant, belonging to the family Polygonaceae. Within the cells of its leaves P. tinctorium accumulates large amounts of a colorless glycoside, indican (indoxyl beta-d-glucoside), from which the blue dye indigo is synthesized. P. tinctorium is well-known in Japan, where it had been cultivated to produce natural indigo for textile dyeing, whereas it represents a potentially interesting new crop in Europe. To better understand the effects of environmental parameters on P. tinctorium crop production and indigo yield, field experiments were carried out in central Italy under temperate climate. Three lines were tested during the 2001 and 2002 growing seasons, and plant/leaf yields as well as indican contents were evaluated. The results showed that P. tinctorium grown in temperate climate conditions can be harvested three times a year. Yields of 82 and 120 t ha(-1) of fresh plant yield were obtained in 2001 and 2002, respectively. The contrasting weather conditions between the two years significantly affected biomass production, which was higher in the 2002 season, characterized by wet weather conditions. The cycle length from sowing to the last harvest was accomplished in 229-238 days when plants had accumulated 2017-2018 degrees C. Green leaves accounted for 40-45% by weight of fresh plant tissue and contained 11-20 g kg(-1) indican. The three lines did not significantly differ in the main productive parameters or in fresh leaf indican content (14.1 g kg(-1) mean value). Photosynthetic active radiation influences indican leaf production according to the model y = 0.0004x + 8.566 (P < 0.01, correlation coefficient = 0.818). Indican content ranged from 12 to 25 g kg(-1) of fresh leaves with PAR daily values from 10000 to 40000 mEinstein m(-2) (recorded in May and at the end of July-beginning of August, respectively). The results indicate that in nonlimiting rainfall conditions a very high indican content and a potentially high indigo yield can be obtained by cultivating P. tinctorium in this pioneer geographical area.

Climate↗
PubMed1985

Indican interference with six commercial procedures for measuring total bilirubin.

We have studied the effect of indican on six commercial procedures for the measurement of total bilirubin in serum. Total bilirubin measured by the Bilirubin A-Gent (Abbott) 2,4-dichlorophenyl diazonium procedure increased by 50 mg/L for each 1 mmol/L of added indican. Similarly, total bilirubin measured by the Bilirubin C-System (Boehringer Mannheim) 2,5-dichlorophenyl diazonium procedure increased by 33 mg/L per mmol/L of indican. Indican also interfered with the Micro Bilirubin Reagent Set (Harleco) Malloy-Evelyn procedure, but to a much lesser extent. The Jendrassik Bilirubin Reagent System (American Monitor) and a modified Jendrassik-Grof procedure (Hoffmann-LaRoche) adapted to the Cobas Bio analyzer were unaffected by the presence of indican. The amount of interference with the 2,5-dichlorophenyl diazonium procedure increased significantly with color development time and was twice the initial amount after 30 min. Concentrations of indican as high as 0.38 mmol/L have been found in sera of patients with renal failure, which would increase total bilirubin values measured by the first two procedures above by 19 and 12 mg/L, respectively. Users of these procedures should therefore be suspicious of unexpectedly high bilirubin values obtained with sera from patients with chronic renal disease.

Autoanalysis↗
PubMed1976

Value of urinary simple phenol and indican determinations in the diagnosis of the stagnant loop syndrome.

The urinary excretion of phenol, p-cresol, and indican was determined in 7 patients with the stagnant loop syndrome, 26 patients with coeliac disease, chronic pancreatitis, and partial gastrectomy, and 18 control patients. The mean excretion of the compounds in the patients with the stagnant loop syndrome and in the control patients, respectively, was 77 and 2.3 mg/24 h of phenol (p less than 0.05), 164 and 39.5 mg/24 h of p-cresol (n.s.), and 369 and 41.5 mg/24 h of indican (p less than 0.01). When applied as diagnostic tests for the stagnant loop syndrome, the phenol excretion showed 2 false negative results, the p-cresol excretion 3 false negative and 2 false positive results, and the indican excretion 6 false positive results. The combined use of phenol and indican determinations eliminated the number of false positive results with the indican test, and was found most useful as screening procedure. Determination of phenol and indican in a 24-hour urine sample is likely to provide a simple method for selecting patients with signs of abnormal bacterial colonization in the small intestine for more detailed investigations.

Aged↗
PubMed1986

Trypsin inhibitor ingestion-induced urinary indican excretion and pancreatic acinar cell hypertrophy.

Sodium saccharin (NaSacc) has been shown to be a protease inhibitor and to induce an increase in urinary indican, which is a product that is dependent on microbial metabolism of tryptophan. These findings suggest that urinary indican might provide a noninvasive marker of increased pancreatic acinar cell size associated with plant trypsin inhibitor ingestion. The results demonstrate the 7.5% of dietary NaSacc, which increases urinary indican, also increases relative pancreas mass (g/kg body weight), and that these effects are not induced by intravenous infusion of NaSacc. Dietary soybean trypsin inhibitor in the dose range of 17-713 mg/100 g diet was associated with parallel dose-dependent increases in urinary indican and pancreatic acinar cell size (assessed histologically). These findings suggest that measurement of relative urinary indican excretion (microgram/g diet ingested) can provide a noninvasive marker of increased pancreatic acinar cell size in rats that ingest compounds which inhibit digestive proteases.

Animals↗
PubMed2007

Formation of the indigo precursor indican in genetically engineered tobacco plants and cell cultures.

The production of the blue dye indigo in plants has been assumed to be a possible route to the introduction of novel coloration into flowers or fibres. As the human cytochrome P450 mono-oxygenase 2A6 (CYP2A6) can form indigo in bacterial cultures, we investigated whether the expression of the corresponding cDNA in transgenic plants could lead to indigo formation. In a first attempt, we generated tobacco cell suspension cultures expressing the cDNA encoding human CYP2A6. Supplementation of the medium with indole led to the generation of indican (3-hydroxyindole-beta-d-glucoside), a metabolite usually exclusively present in indigoferous dye plants. Hence, the recombinant CYP2A6 converted indole to the reactive metabolite 3-hydroxyindole (indoxyl), whereas rapid glucosylation is obviously conducted by ubiquitous plant glucosyl transferases (GTs). Interestingly, of nine additionally tested plant cell suspension cultures from various plant families, five were also capable of the formation of indican after indole supplementation, although this metabolism was more pronounced in transgenic tobacco cell suspension cultures expressing CYP2A6 cDNA. To evaluate whether indican or even indigo could be produced in whole plants, we generated transgenic tobacco plants harbouring active CYP2A6 together with an indole synthase (BX1) from maize. The genetically engineered tobacco plants accumulated indican, but did not develop a blue coloration. Although the de novo formation of indican in transgenic tobacco plants hampered indigo formation, it supports the contention that biosynthetic pathways can be efficiently mimicked by metabolic engineering.

Aryl Hydrocarbon Hydroxylases↗
PubMed1983

Indican excretion in the elderly.

It has been suggested that up to a half of elderly hospitalized patients excrete an abnormally high concentration of indican compared with young individuals. Thus, the commonly used colorimetric qualitative test for urinary indican excess may be invalid in the aged. However, of 21 young control subjects, 18 healthy elderly subjects, and 23 disabled, elderly, long-stay patients, no individual had levels of indican detectable by the standard colorimetric qualitative test. The use of a quantitative assay demonstrated that urinary indican concentration is similar in young and elderly individuals. A diurnal variation in urinary indican concentration was discovered which may affect the interpretation of the qualitative test.

Adult↗
PubMed1985

A 2,4-dichlorophenyl diazonium-based method for total bilirubin without interference from indican in uremic sera.

Comparison of total bilirubin quantification by a 2,4-dichlorophenyl diazonium method (2,4-DCPD) with a Jendrassik-Grof type of method showed excellent correlation for randomly selected sera. However, sera from uremic patients on chronic hemodialysis showed a marked positive bias for the 2,4-DCPD result as compared with the Jendrassik-Grof result. The mean difference was 5.3 mg/L, and resulted in about 20% of the hemodialysis patients having bilirubin values greater than 13 mg/L, the upper limit of our reference range. Indican in uremic sera reportedly reacts with certain diazo reagents, so we investigated indican's reactivity in the above methods. In vitro addition of indican caused no interference in the Jendrassik-Grof method, but produced a significant positive interference in the 2,4-DCPD method, 1 mmol of indican per liter appearing as about 36 mg of total bilirubin per liter. Long reaction times with the 2,4-DCPD reagent accentuate the problem. By shortening the reaction time with the 2,4-DCPD reagent to 1.7 min, we find that the indican interference can be eliminated, without affecting quantification of total bilirubin in either normal or uremic sera.

Bilirubin↗