Efficient biotransformation of non-steroid anti-inflammatory drugs by endophytic and epiphytic fungi from dried leaves of a medicinal plant, Plantago lanceolata L.

In the current study, decomposition of diclofenac, diflunisal, ibuprofen, mefenamic acid and piroxicam was tested using nine identified strains of endophytic and epiphytic fungi (from Ascomycota) adapted to natural products resembling the pharmaceuticals. The strains were isolated from a medicinal plant, Plantago lanceolata leaves. Metabolites were tentatively identified by liquid chromatography - tandem mass spectrometry (LC-MS3). Eighteen of the 45 combinations resulted in significant decrease of the concentration of the NSAIDs in model solutions. The most active strains were Aspergillus nidulans and Bipolaris tetramera, while Epicoccum nigrum and Aspergillus niger showed somewhat less potency. Piroxicam and diclofenac were most resistant to biotransformation, while ibuprofen and mefenamic acid were efficiently metabolized by most strains. Ten metabolites could be tentatively identified, including hydroxy-metabolites of all tested NSAIDs, and a dihydroxy-metabolite of piroxicam. This biotransformation is likely to modify the toxicity and bioaccumulation potential of these pharmaceuticals. The results highlight the applicability of polyphenol-rich dried medicinal plant materials as an excellent source of fungi with high biotransforming potential. The results also suggest more in-depth testing of these fungi for biodegradation processes

Myrosinase Compatible Simultaneous Determination of Glucosinolates and Allyl Isothiocyanate by Capillary Electrophoresis Micellar Electrokinetic Chromatography (CE-MEKC)

Introduction. The functional food Cruciferous vegetables contain glucosinolates which are decomposed by the myrosinase enzyme upon tissue damage. The isothiocyanates are the most frequent decomposition products. Because of their various bioactivities, these compounds and the myrosinase is of high interest to many scientific fields. Objective. Development of a capillary electrophoresis method capable of myrosinase- compatible, simultaneous quantification of glucosinolates and isothiocyanates. Methods. Capillary electrochromatography parameters were optimized, followed by optimization of a myrosinase-compatible derivatization procedure for isothiocyanates. Vegetable extracts (Brussels sprouts, horseradish, radish and watercress) were tested for myrosinase activity, glucosinolate content and isothiocyanate conversion rate. Allyl isothiocyanate was quantified in some food products. Results. The method allows quantification of sinigrin, gluonasturtiin and allyl isothiocyanate after myrosinase compatible derivatization in-vial by mercaptoacetic acid. The chromatograhpic separation takes 2.5 minutes (short end injection) or 15 minutes (long end injection). For the tested vegetables, measured myrosinase activity was between 0.960 – 27.694 and 0.461 – 26.322 μmol min-1 mg-1 protein, glucosinolate content was between 0 – 2291.8 and 0 – 248.5 μg g-1 fresh weight for sinigrin and gluconastrutiin, respectively. The possible specificity of plants to different glucosinolates was also shown. Allyl isothiocyanate release rate was different in different vegetables (73.13 to 102.13 %). The method could also be used for quantification of allyl isothiocyanate from food products. Conclusions. The presented capillary electrophoresis method requires a minimal amount of sample and contains only a few sample preparation steps, and can be used in several applications (glucosinolate determination, myrosinase activity measurement, isothiocyanate release estimation)

A Simple Method for On-Gel Detection of Myrosinase Activity

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Chemotyping of terrestrial Nostoc-like isolates from alkali grassland areas by non-targeted peptide analysis

The Nostoc genus is a well-known heterocytous, filamentous cyanobacterium which can be found all over the world. The size of terrestrial and/or freshwater colonies can be microscopic and macroscopic as well. In addition, Nostoc species are one of the most common photosynthetic cyanobacterial partners in symbiotic interactions. Terrestrial cyanobacterial colonies were collected and isolated in this study from various alkali grassland habitats (Great Hungarian Plain). Altogether 133 colonies were isolated from the 65 collected samples. The peptide patterns of the Nostoc-like strains were examined using HPLC-ESI-MS/MS and 41 peptides were identified from 45 isolated Nostoc-like strains; these compounds belonged to 4 different peptide classes. Twelve nostoginin/microginin, 16 anabaenopeptin, 12 banyaside/suomilide variants were identified. 37% of our isolated Nostoc-like strains produced some of the peptide metabolites we tested. These strains showed distinct chemotypes according to their peptide patterns, and can be divided into 4 groups based on their metabolisms. Strains either contained: (1) nostoginins/microginins, (2) anabaenopeptins, (3) anabaenopeptins and banyasides or (4) banyasides as major compounds. Banyasides were present in many of our strains and showed very high intensity in some cases. A number of previously unknown banyaside variants have been identified

Allyl-isothiocyanate and microcystin-LR reveal the protein phosphatase mediated regulation of metaphase-anaphase transition in Vicia faba

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