Glucosinolate Hydrolysis Products Research Articles

Initialin vitroevaluations of the antibacterial activities of glucosinolate enzymatic hydrolysis products against plant pathogenic bacteria

The aim of the study was to evaluate the in vitro antibacterial effects of glucosinolate hydrolysis products (GHP) against plant pathogenic micro-organisms namely Agrobacterium tumefaciens, Erwinia chrysanthemi, Pseudomonas cichorii, Pseudomonas tomato, Xanthomonas campestris and Xanthomonas juglandis. Using a disc diffusion assay, seven different doses of 10 GHP were tested against each bacteria. The results showed that the isothiocyanates were potent antibacterials, whilst the other GHP were much less efficient. Moreover, the antibacterial effects were dose-dependent, increasing with the dose applied; 2-phenylethylisothiocyanate and sulforaphane showed the strongest inhibitory effects. The overall results show a great potential for using the isothiocyanates as an alternative tool to control undesired bacterial growth in plants. Glucosinolate hydrolysis products and more specifically the isothiocyanates: benzylisothiocyanate, 2-phenylethylisothiocyanate, the isothiocyanate Mix and sulforaphane, were effective phytochemicals against the in vitro growth of the phytopathogenic bacteria. The antibacterial activity exhibited by these phytochemicals reinforces their potential as alternatives to the traditional chemical control of phytopathogenic bacteria. This current in vitro study is the first providing comparative data on GHP as potential control agents for plant pathogenic bacteria. However, more studies are needed to determine their possible allelopathic impacts e.g. inhibition of plant growth and negative effects on beneficial soil bacteria and fungi (mycorrhizae).

The antimicrobial effects of glucosinolates and their respective enzymatic hydrolysis products on bacteria isolated from the human intestinal tract

The aim of the study was to evaluate the in vitro antibacterial activity of glucosinolates and their enzymatic hydrolysis product against bacteria isolated from the human intestinal tract. Using a disc diffusion bioassay, different doses of intact glucosinolates and their corresponding hydrolysis products were tested. There were clear structure-activity and concentration differences with respect to the in vitro growth inhibition effects as well as differences in the sensitivities of the individual bacteria. The most effective glucosinolate hydrolysis products were the isothiocyanates; sulforaphane and benzyl isothiocyanate were the best inhibitors of growth. Indole-3-carbinol had some inhibitory effects against the Gram-positive bacteria but had no effect, even at the highest dose, against the Gram-negative bacteria. Indole-3-acetonitrile had some inhibitory activity against the Gram-negative bacteria. Glucosinolates, nitriles and amines were ineffective at all the doses used. Glucosinolate hydrolysis products and specifically the isothiocyanates SFN and BITC have significant antimicrobial activity against Gram-positive and Gram-negative bacteria, and might be useful in controlling human pathogens through the diet. This the first major in vitro study demonstrating the potential of these natural dietary chemicals as an alternative to, or in combination with, current antibiotic-based therapies for treating infectious diseases.

Cruciferous vegetable intake and the risk of human cancer: epidemiological evidence

Over several decades a number of epidemiological studies have identified the inverse associations between cruciferous vegetables and the risk of several cancers, including gastric, breast, colo-rectal, lung, prostate, bladder and endometrial cancers, via plausible physiological mechanisms. Although retrospective case–control studies have consistently reported inverse associations between the risk of these cancers and the intake of cruciferous vegetables and isothiocyanate-containing plants, current prospective cohort studies have found these associations to be weaker and less consistent. Genetic variations affecting the metabolism of glucosinolate hydrolysis products may modulate the effects of cruciferous vegetable consumption on cancer risk, which may be one of the reasons for the discrepancies between retrospective and prospective studies. In addition, methodological issues such as measurement errors of dietary exposure, misclassification, recall bias, publication bias, confounding and study design should be carefully considered in interpreting the results of case–control and cohort studies and in drawing conclusions in relation to the potential effects of cruciferous vegetables on cancers. Although recent comprehensive reviews of numerous studies have purported to show the specific protective role of cruciferous vegetables, and particularly Brassicas, against cancer risk, the current epidemiological evidence suggests that cruciferous vegetable consumption may reduce the risk only of gastric and lung cancers. However, there is at present no conclusive evidence that the consumption of cruciferous vegetables attenuates the risk of all other cancers.

The ‘mustard oil bomb’: not so easy to assemble?! Localization, expression and distribution of the components of the myrosinase enzyme system

Glucosinolates are plant secondary metabolites that are hydrolysed by the action of myrosinases into various products (isothiocyanates, thiocyanates, epithionitriles, nitriles, oxazolidines). Massive hydrolysis of glucosinolates occurs only upon tissue damage but there is also evidence indicating metabolism of glucosinolates in intact plant tissues. It was originally believed that the glucosinolate–myrosinase system in intact plants was stable due to a spatial separation of the components. This has been coined as the ‘mustard oil bomb’ theory. Proteins that form complexes with myrosinases have been described: myrosinase-binding proteins (MBPs) and myrosinase-associated proteins (MyAPs/ESM). The roles of these proteins and their biological relevance are not yet completely known. Other proteins of the myrosinase enzyme system are the epithiospecifier protein (ESP) and the thiocyanate-forming protein (TFP) that divert the glucosinolate hydrolysis from isothiocyanate production to nitrile/epithionitrile or thiocyanate production. Some glucosinolate hydrolysis products act as plant defence compounds against insects and pathogens or have beneficial health effects on humans. In this review, we survey and critically assess the available information concerning the localization, both at the tissular/cellular and subcellular level, of the different components of the myrosinase enzyme system. Data from the model plant Arabidopsis thaliana is compared to that from other glucosinolate-producing Brassicaceae in order to show common as well as divergent features of the ‘mustard oil bomb’ among these species.

Regulation and function of specifier proteins in plants

Specifier proteins are responsible for the diversification of biologically active products formed upon myrosinase-catalyzed glucosinolate hydrolysis and are therefore assumed to have an impact on the defensive function of the glucosinolate–myrosinase system. Among glucosinolate hydrolysis products, the generation of epithionitriles and organic thiocyanates requires the presence of epithiospecifier protein (ESP) and thiocyanate-forming protein (TFP), respectively, while myrosinase alone is sufficient for the production of isothiocyanates. Both ESP and TFP also promote the formation of simple nitriles upon myrosinase-catalyzed glucosinolate hydrolysis. Only little is known about the biological effects of epithionitriles and thiocyanates. Moreover, simple nitriles have repeatedly been reported to be less toxic to plant pathogens and herbivorous insects than the correponding isothiocyanates. Thus, it has remained an open question how plants benefit from the presence of specifier proteins. In this review, we survey the biological effects of different types of glucosinolate hydrolysis products on insects and pathogens as well as the current knowlegde on the developmental, organ specific and stimuli-mediated regulation of specifier proteins. Integrating these findings can help us to better understand the ecological functions of plant specifier proteins as well as the co-evolution of glucosinolate-containing plants and their insect herbivores.

Transcriptional and biochemical signatures of divergence in natural populations of two species of New Zealand alpinePachycladon

New Zealand is diverse in alpine and subalpine environments, a consequence of Late Tertiary tectonic and climatic change. However, few studies have sought to evaluate the importance of these environments as abiotic drivers in the diversification of plant species. Of particular interest is the Late Tertiary radiation of Pachycladon, an endemic New Zealand genus of alpine cress. Here we report observations on genome-wide levels of differential expression measured in the habitats of two closely related species of Pachycladon with distinct altitudinal preferences. Using Arabidopsis microarrays, we have identified 310 predominantly hormone- and stress-response genes up-regulated in Pachycladon fastigiata and 324 genes up-regulated in Pachycladon enysii. Expression patterns for glucosinolate biosynthesis and hydrolysis genes (MAM1, MAM-I, MAM-D, AOP2, ESP, ESM1) as well as flavonoid biosynthesis genes (F3'H, FLS, FAH1) were found to be species specific. Predicted differences in flavonoid contents were partly confirmed by high performance liquid chromatography analysis. Differences in glucosinolate profiles and glucosinolate hydrolysis products obtained by high performance liquid chromatography and gas chromatography-mass spectrometry analysis, respectively, also supported inferences from expression analyses. Five glucosinolate chemotypes were matched to known Arabidopsis ecotypes, and the potential adaptive significance of these chemotypes has been discussed. Our findings, in contrast to expectations for evolution of the New Zealand flora, suggest that biotic drivers, such as plant-herbivore interactions, are likely to be as important as abiotic drivers in the diversification of Pachycladon.

Comparative Innate Responses of the Aphid Parasitoid Diaeretiella rapae to Alkenyl Glucosinolate Derived Isothiocyanates, Nitriles, and Epithionitriles

Cruciferous plants (Brassicaceae) are characterized by the accumulation of a group of secondary metabolites known as glucosinolates that, following attack by pathogens or herbivores, may be hydrolyzed to one of a number of products including isothiocyanates and nitriles. Despite the range of hydrolysis products that may be produced, the toxicity of glucosinolates to pathogens and herbivores may be explained largely by the production of isothiocyanates. Isothiocyanates are also known to provide an indirect defense by acting as host finding cues for parasitoids of insect herbivores that attack crucifers. It has been speculated that nitriles may provide a similar indirect defense. Here, we investigate the olfactory perception and orientation behavior of the aphid parasitoid Diaeretiella rapae, to a range of alkenylglucosinolate hydrolysis products, including isothiocyanates, nitriles, and epithionitriles. Electroantennogram responses indicated peripheral odor perception in D. rapae females to all 3-butenylglucosinolate hydrolysis products tested. By contrast, of the 2-propenylglucosinolate hydrolysis products tested, only the isothiocyanate elicited significant responses. Despite showing peripheral olfactory detection of a range of 3-butenylglucosinolate hydrolysis products, naïve females oriented only to the isothiocyanate. Similarly, parasitoids oriented to 3-isothiocyanatoprop-1-ene, but not to the corresponding nitrile or epithionitrile. However, by rearing D. rapae either on Brassica nigra, characterized by the accumulation of 2-propenylglucosinolate, or Brassica rapa var rapifera, characterized by the accumulation of 3-butenylglucosinolate, altered the innate response of parasitoids to 3-isothiocyanatoprop-1-ene and 4-isothiocyanatobut-1-ene. These results are discussed in relation to the defensive roles of glucosinolate hydrolysis products and the influence of the host plant on aphid parasitoid behavior.

Glucosinolates and biofumigation: fate of glucosinolates and their hydrolysis products in soil

The bioactive hydrolysis products of glucosinolates, particularly the isothiocyanates, can be used to control soil pests and weeds by incorporating glucosinolate-containing plant material in soil—a practice known as biofumigation. The fate of glucosinolates and their hydrolysis products in soil determines both the efficacy and environmental impact of biofumigation. Knowledge of the processes by which these compounds are sorbed, degraded or otherwise lost from the soil is fundamental to developing effective, but environmentally benign biofumigation strategies. Effective biofumigation relies on maximum hydrolysis of the glucosinolate in the plant tissue to generate high isothiocyanate concentrations in the soil after incorporation. This is favoured by maximum cell disruption, by addition of water, and a high soil temperature. Residual glucosinolates are very weakly sorbed, readily leached and are microbially degraded and mineralised in soil. In contrast, isothiocyanates are strongly sorbed by the organic matter in soil, react strongly with nucleophilic groups present in soil, and are prone to volatilization losses in addition to microbial degradation and mineralisation. These loss processes are influenced by soil type, water content and temperature. Using appropriate incorporation strategies, sufficiently high isothiocyanate concentrations (>100 nmol g−1) can be achieved in soil using biofumigation for effective suppression of susceptible pests. The relatively rapid sorption and degradation of the isothiocyanates in the period of days after incorporation minimizes the risks of persistence in the environment or leaching. Biofumigation is therefore a promising technique which can be further developed to form part of IPM (Integrated Pest Management) strategies to reduce reliance on synthetic pesticides with minimal unintended impacts on the environment.

The Desert Locust, Schistocerca gregaria, Detoxifies the Glucosinolates of Schouwia purpurea by Desulfation

Desert locusts (Schistocerca gregaria) occasionally feed on Schouwia purpurea, a plant that contains tenfold higher levels of glucosinolates than most other Brassicaceae. Whereas this unusually high level of glucosinolates is expected to be toxic and/or deterrent to most insects, locusts feed on the plant with no apparent ill effects. In this paper, we demonstrate that the desert locust, like larvae of the diamondback moth (Plutella xylostella), possesses a glucosinolate sulfatase in the gut that hydrolyzes glucosinolates to their corresponding desulfonated forms. These are no longer susceptible to cleavage by myrosinase, thus eliminating the formation of toxic glucosinolate hydrolysis products. Sulfatase is found throughout the desert locust gut and can catalyze the hydrolysis of all of the glucosinolates present in S. purpurea. The enzyme was detected in all larval stages of locusts as well as in both male and female adults feeding on this plant species. Glucosinolate sulfatase activity is induced tenfold when locusts are fed S. purpurea after being maintained on a glucosinolate-free diet, and activity declines when glucosinolates are removed from the locust diet. A detoxification system that is sensitive to the dietary levels of a plant toxin may minimize the physiological costs of toxin processing, especially for a generalist insect herbivore that encounters large variations in plant defense metabolites while feeding on different species.

Transcriptional responses of Arabidopsis thaliana ecotypes with different glucosinolate profiles after attack by polyphagous Myzus persicae and oligophagous Brevicoryne brassicae

Plants are equipped with a range of defence mechanisms against herbivorous insects. In cruciferous species, jasmonic acid, salicylic acid, and ethylene along with glucosinolates and their hydrolysis products play important roles in plant protection and plant-insect communication. In turn, a number of herbivores have adapted to plants that contain glucosinolates. As a result of adaptation to their host plants, specialized insects may elicit different plant-inducible responses than generalists. Oligonucleotide microarrays and qRT-PCR analysis were used to characterize transcriptional profiles of Arabidopsis thaliana plants in response to infestation with a generalist aphid, Myzus persicae, or a cruciferous plant specialist, Brevicoryne brassicae. To find possible differences and similarities in molecular responses between plants differing in predominant glucosinolate hydrolysis products, three ecotypes of A. thaliana were chosen: Wassilewskija (Ws), Cape Verde Islands (Cvi), and Landsberg erecta (Ler), which, respectively, produce mainly isothiocyanates, epithionitriles, and nitriles. In all three ecotypes, general stress-responsive genes, genes belonging to octadecanoid and indole glucosinolate synthesis pathways were induced upon both generalist and specialist attack. By contrast, transcription of myrosinases, enzymes hydrolysing glucosinolates, was suppressed. The induction of the jasmonic acid synthesis pathway was strongest in Cvi, while the up-regulation of the indole glucosinolate synthesis pathway was highest in Ler, suggesting a slightly different defence strategy in these two ecotypes. Specialist and generalist infestations caused statistically significant differential regulation of 60 genes in Ws and 21 in Cvi. Among these were jasmonic acid and tryptophan synthesis pathway enzymes, and pathogenesis related protein (PR1). Insect no-choice experiments revealed lowered fitness of B. brassicae on Ler and Cvi in comparison to Ws, but no ecotype-dependent change in fecundity of M. persicae. Targeted studies employing constructs of GUS reporter gene under the control of promoters from CYP79B2 and CYP79B3 genes showed insect-specific induction of the indole glucosinolates synthesis pathway.

Glucosinolate hydrolysis in Lepidium sativum––identification of the thiocyanate-forming protein

Glucosinolates are a class of thioglycosides found predominantly in plants of the order Brassicales whose function in anti-herbivore defense has been attributed to the products formed by myrosinase-catalyzed hydrolysis upon plant tissue damage. The most common type of hydrolysis products, the isothiocyanates, are toxic to a wide range of organisms. Depending on the glucosinolate side-chain structure and the presence of certain protein factors, other types of hydrolysis products, such as simple nitriles, epithionitriles and organic thiocyanates, can be formed whose biological functions are not well understood. Of the proteins controlling glucosinolate hydrolysis, only epithiospecifier proteins (ESPs) that promote the formation of simple nitriles and epithionitriles have been identified on a molecular level. We investigated glucosinolate hydrolysis in Lepidium sativum and identified a thiocyanate-forming protein (TFP) that shares 63-68% amino acid sequence identity with known ESPs and up to 55% identity with myrosinase-binding proteins from Arabidopsis thaliana, but differs from ESPs in its biochemistry. TFP does not only catalyze thiocyanate and simple nitrile formation from benzylglucosinolate but also the formation of simple nitriles and epithionitriles from aliphatic glucosinolates. Analyses of glucosinolate hydrolysis products in L. sativum autolysates and TFP transcript accumulation revealed an organ-specific regulation of thiocyanate formation. The identification of TFP defines a new family of proteins that control glucosinolate hydrolysis and challenges the previously proposed reaction mechanism of epithionitrile formation. As a protein that promotes the formation of a wide variety of hydrolysis products, its identification provides an important tool for further elucidating the mechanisms of glucosinolate hydrolysis as well as the ecological role and the evolutionary origin of the glucosinolate-myrosinase system.

Altered Glucosinolate Hydrolysis in Genetically Engineered Arabidopsis thaliana and its Influence on the Larval Development of Spodoptera littoralis

The antiherbivore potential of the glucosinolate-myrosinase defense system found in plants of the order Capparales is heavily influenced by the types of hydrolysis products (e.g. isothiocyanates, nitriles) formed from the parent glucosinolates upon plant damage. However, comparison of the effects of glucosinolate hydrolysis products on insect herbivores has been hampered by the lack of suitable experimental tools for rigorous bioassays, such as intact plants differing only in the types of hydrolysis products they produce, or artificial diets that can accurately simulate glucosinolate hydrolysis. The wide array of molecular resources for Arabidopsis thaliana has facilitated the identification of several genes that play a role in glucosinolate hydrolysis. One of these encodes the epithio-specifier protein (ESP) that promotes the formation of nitriles at the expense of isothiocyanates in certain ecotypes of A. thaliana. We overexpressed the ESP cDNA from the nitrile-producing ecotype Landsberg erecta in the isothiocyanate-producing ecotype Columbia-0 to generate transgenic lines of A. thaliana that differed from wild-type plants in the type of glucosinolate hydrolysis products formed upon tissue damage, whereas parent glucosinolate profile and myrosinase activity levels, as well as plant morphology and growth habit, remained unchanged. Bioassays with the model generalist herbivore Spodoptera littoralis (Lepidoptera: Noctuidae) demonstrated that larvae reared on the nitrile-producing lines on average gained weight faster in the first larval stages than larvae that fed on isothiocyanate-producing control plants. Furthermore, larvae with medial growth rates showed a tendency to pupate earlier on the ESP-overexpressing plant lines. Together with the results of previous studies, these findings suggest that isothiocyanates are more effective defenses against insect herbivores than nitriles, and raise questions about what conditions select for nitrile formation in plants.

Association between Consumption of Cruciferous Vegetables and Condiments and Excretion in Urine of Isothiocyanate Mercapturic Acids

A high intake of cruciferous vegetables is associated with a reduced risk of cancer and cardiovascular diseases. This protective effect has been linked to isothiocyanates, enzymatic hydrolysis products of glucosinolates. In this study, the metabolic fate of glucosinolates and isothiocyanates after ingestion of 19 different cruciferous vegetables was studied in three male subjects. After the consumption of 13 cruciferous vegetables (glucosinolate content, 0.01-0.94 mmol/kg) and six condiments (isothiocyanate content, 0.06-49.3 mmol/kg), eight different isothiocyanate mercapturic acids were determined in urine samples. Excretion levels after the consumption of raw vegetables and condiments were higher (bioavailability, 8.2-113%) as compared to cooked vegetables (bioavailability, 1.8-43%), but the excretion rate was similar (t1/2=2.1-3.9 h). Isothiocyanates in urine remain longer at a nonzero level after the consumption of glucosinolates from cooked vegetables, as compared to raw vegetables and condiments, and maximal levels in urine were reached about 4 h later. Isothiocyanate mercapturic acids can be used as a biomarker to reflect the active dose of isothiocyanates absorbed.

Micellar electrokinetic capillary chromatography—Synchronous monitoring of substrate and products in the myrosinase catalysed hydrolysis of glucosinolates

A micellar electrokinetic capillary chromatography (MECC) method has been developed for monitoring the myrosinase catalysed hydrolysis of 2-hydroxy substituted glucosinolates and the simultaneous formation of the corresponding degradation products (oxazolidine-2-thiones (OZTs) and nitriles). Glucosibarin ((2 R)-2-hydroxy-2-phenylethylglucosinolate) was chosen as the model glucosinolate owing to the difficulties in determining hydrolysis rates of this type of substrates in traditional UV-assays. The method was afterwards validated with glucobarbarin ((2 S)-2-hydroxy-2-phenylethylglucosinolate) and progoitrin ((2 R)-2-hydroxybut-3-enylglucosinolate). Aromatic glucosinolates without a 2-hydroxy group in their side chains, such as glucotropaeolin (benzylglucosinolate) and gluconasturtiin (phenethylglucosinolate) were also tested. Formation of the glucosinolate hydrolysis products was monitored simultaneously at 206 nm and 230 nm. This allowed estimation of the extinction coefficient of the OZT derived from glucosibarin, which was found to be 18,000 M −1 cm −1 and 12,000 M −1 cm −1 at 206 nm and 230 nm, respectively. The developed method has limit of detection of 0.04 mM and 0.06 mM and limit of quantification of 0.2 mM and 0.3 mM for the glucosibarin derived OZT and nitrile, respectively. Linearity of the glucosinolate concentration was examined at six concentration levels from 2.5 mM to 100 mM and at 206 nm a straight line ( R 2 = 0.9996) was obtained. The number of theoretical plates ( N) at the optimal system conditions was 245,000 for the intact glucosibarin, 264,000 for the OZT and 252,000 for the nitrile.

Production of glucosinolate hydrolysis products in Farsetia aegyptia suspension cultures following elicitation

Levels of glucosinolates in Farsetia aegyptia var. ovalis suspension cultures were monitored after treatment with yeast extract, chitosan, methyl jasmonate, ampicillin, and Phytophthora infestans autoclaved mycelia as elicitors. Glucosinolates were identified, and an estimation of their levels obtained from their hydrolysis products. Yeast extract improved glucotropaeolin (benzyl-glucosinolate) and glucocheirolin [3-(methylsulfonyl)propyl-glucosinolate] accumulation, and production of sec-butyl, isobutylglucosinolate and gluconasturtiin (2-phenylethyl-glucosinolate) was only detected in yeast elicited cultures. Increases were shown in glucotropaeolin levels in cultures elicited using methyljasmonate, and chitosan and methyljasmonate in combination.

Brassica napus seed meal soil amendment modifies microbial community structure, nitric oxide production and incidence of Rhizoctonia root rot

A low glucosinolate content (21.8 μmol g −1) Brassica napus seed meal (RSM) applied to orchard soils altered communities of both pathogenic and saprophytic soil micro-organisms. RSM amendment reduced infection by native and introduced isolates of Rhizoctonia spp. and recovery of Pratylenchus spp. from apple roots. Root infection by Rhizoctonia solani AG-5 was also suppressed in split-root assays where a portion of the root system was cultivated in RSM-amended soils and the remainder grown in the presence of the pathogen but lacking RSM. R. solani hyphal growth was not inhibited by RSM amendment. Suppression of Pratylenchus was attained to an equivalent extent by amending soils with either RSM or soybean meal (SM) when applied to provide a similar N content. Thus, glucosinolate hydrolysis products did not appear to have a significant role in the suppression of Rhizoctonia spp. or Pratylenchus spp. obtained via RSM amendment. RSM amendment elevated populations of Pythium spp. and of ammonia-oxidizing bacteria that release nitric oxide but suppressed fluorescent pseudomonad numbers. Streptomyces spp. soil populations increased significantly in response to RSM but not SM amendment. The vast majority of Streptomyces spp. recovered from the apple rhizosphere produced nitric oxide and possessed a nitric oxide synthase homolog. We propose that transformations in the bacterial community structure are associated with the observed control of Rhizoctonia root rot, with NO production by soil bacteria potentially having a role in the induction of plant systemic resistance.

Glucosinolate hydrolysis products from various plant sources: pH effects, isolation, and purification

Glucosinolates are a class of organic anions that can be hydrolyzed either enzymatically with myrosinase or non-enzymatically to form primarily isothiocyanates and/or nitriles. The isolation and purification of these hydrolysis products are of particular interest both for their potential use in organic synthesis and for their biological activities. Methods were developed for the isolation and purification of gram-scale quantities of several ω-(methylthio)alkyl-, ω-(methylsulfinyl)alkyl-, ω-(methylsulfonyl)alkyl-, and substituted benzyl glucosinolate hydrolysis products; the isothiocyanates erucin [1-isothiocyanato-4-(methylthio)butane], iberin [1-isothiocyanato-3-(methylsulfinyl)propane], cheirolin [1-isothiocyanato-3-(methylsulfonyl)propane], lesquerellin [1-isothiocyanato-6-(methylthio)hexane], hesperin [1-isothiocyanato-6-(methylsulfinyl)hexane], sulforaphene [1-isothiocyanato-4-(methylsulfinyl)but-3-ene], 3-methoxybenzyl isothiocyanate, and 4-hydroxybenzyl isothiocyanate; and the nitriles iberverin nitrile [1-cyano-3-(methylthio)propane], erucin nitrile [1-cyano-4-(methylthio)butane], and 3-methoxybenzyl nitrile using defatted seedmeal from several different genera within the crucifer family (Brassicaceae) and meadowfoam family (Limnanthaceae) as the source of parent glucosinolates. The procedures use solvent extraction of autolyzed defatted seedmeals from various plant sources together with variable reaction pHs and solvent partitioning to obtain relatively pure (generally >97%) compounds without the need for chromatographic separation.

Production of two volatile glucosinolate hydrolysis compounds in Nasturtium montanum and Cleome chelidonii plant cell cultures

Callus and suspension cultures established from Nasturtium montanum and Cleome chelidonii were shown to produce glucosinolates by analysis of their hydrolysis products. Large increases in two glucosinolate hydrolysis products were noted when cultures were supplemented with l-cysteine and l-methionine, and further increases were produced in N. montanum with l-tryptophan supplementation.

Dietary agent indole-3-carbinol protects female rats against the hepatotoxicity of the antitumor drug ET-743 (trabectidin) without compromising efficacy in a rat mammary carcinoma.

ET-743, an experimental antitumor drug with promising activity in sarcoma, breast and ovarian carcinoma, is currently under phase 2 clinical evaluation. It is hepatotoxic in animals and patients. We tested the hypothesis that indole-3-carbinol (I3C), the hydrolysis product of glucosinolates occurring in cruciferous vegetables, may protect against ET-743-induced hepatotoxicity in the female Wistar rat, the animal species with the highest sensitivity toward the adverse hepatic effect of this drug. Hepatotoxicity was adjudged by measurement of plasma levels of bilirubin, alkaline phosphatase (ALP) and aspartate aminotransferase (AST) and by liver histopathology. The effect of I3C on the kinetics of ET-743 in rat plasma and liver was investigated by high-pressure liquid chromatography. The effect of I3C on the antitumor efficacy of ET-743 was explored in rats bearing the 13762 mammary carcinoma. ET-743 (40 microg/kg i.v.) alone caused an elevation of plasma bilirubin, ALP and AST levels and degeneration and patchy focal necrosis of bile duct epithelial cells. Addition of I3C to the diet (0.5%) for 6 days prior to ET-743 administration almost completely abolished manifestations of hepatotoxicity. In contrast, a dietary concentration of 0.1% I3C did not protect, nor did dietary diindolylmethane (0.2%), an acid-catalyzed condensation product of I3C. Ingestion by rats of I3C for 6 days prior to ET-743 (40 microg/kg i.v.) decreased plasma but not hepatic concentrations of ET-743 compared to animals that received ET-743 alone. I3C did not interfere with the antitumor efficacy of ET-743. The results suggest that ingestion of I3C may counteract the unwanted effect of ET-743 in the liver. I3C should be investigated as a hepatoprotectant in patients who receive ET-743 therapy.

Glucosinolate hydrolysis products from various plant sources: pH effects, isolation, and purification*1

Glucosinolates are a class of organic anions that can be hydrolyzed either enzymatically with myrosinase or non-enzymatically to form primarily isothiocyanates and/or nitriles. The isolation and purification of these hydrolysis products are of particular interest both for their potential use in organic synthesis and for their biological activities. Methods were developed for the isolation and purification of gram-scale quantities of several ω-(methylthio)alkyl-, ω-(methylsulfinyl)alkyl-, ω-(methylsulfonyl)alkyl-, and substituted benzyl glucosinolate hydrolysis products; the isothiocyanates erucin [1-isothiocyanato-4-(methylthio)butane], iberin [1-isothiocyanato-3-(methylsulfinyl)propane], cheirolin [1-isothiocyanato-3-(methylsulfonyl)propane], lesquerellin [1-isothiocyanato-6-(methylthio)hexane], hesperin [1-isothiocyanato-6-(methylsulfinyl)hexane], sulforaphene [1-isothiocyanato-4-(methylsulfinyl)but-3-ene], 3-methoxybenzyl isothiocyanate, and 4-hydroxybenzyl isothiocyanate; and the nitriles iberverin nitrile [1-cyano-3-(methylthio)propane], erucin nitrile [1-cyano-4-(methylthio)butane], and 3-methoxybenzyl nitrile using defatted seedmeal from several different genera within the crucifer family (Brassicaceae) and meadowfoam family (Limnanthaceae) as the source of parent glucosinolates. The procedures use solvent extraction of autolyzed defatted seedmeals from various plant sources together with variable reaction pHs and solvent partitioning to obtain relatively pure (generally >97%) compounds without the need for chromatographic separation.