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      Lentinula edodes Mycelium as Effective Agent for Piroxicam Mycoremediation

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          Abstract

          Pollution of the environment with inorganic and organic substances is one of the main problems in the world. For this reason, it is necessary to conduct researches for effective methods of biodegradation of xenobiotics, including drugs whose unmetabolized forms are introduced into the environment, especially into water. One possible solution to this problem may be the use of white rot fungi, such as Lentinula edodes. This is an edible species used in medicine because of its beneficial anti-cancer, hypocholesterolemic, hypotensive, hypoglycemic and antioxidant effects. Due to the fact that the mycelium of L. edodes produces enzymes with oxidizing properties that can degrade xenobiotics. The aim of the work was verification if in vitro cultures of L. edodes can be used for bioremediation of non-steroidal, anti-inflammatory drug: piroxicam. For this purpose, the in vitro culture of L. edodes was derived and the mycelial cultures of this species enriched with piroxicam were analyzed. The biodegradation pathway of piroxicam by L. edodes mycelium was carried out by the UPLC/MS/MS method. The degradation process of piroxicam was found to affect primarily the linker between the thiazine and the piperidine ring, leading to its oxidation and cleavage. Additionally, oxidation of the benzothiazine moiety was observed, leading to hydroxylation and oxidation of the phenyl ring as well as oxidation of the thiazine ring leading to partial or complete removal of the sulfonamide moiety. It seems that the degradation process led finally to 2-hydroxybenozquinone, which may be further oxidized to inorganic compounds. What’s more, concentration of piroxicam in in vitro cultures of L. edodes was not correlated with effectiveness of biodegradation of this compound – on each experimental series, the level of degradation was the same. The results confirm the possibility of using the investigated L. edodes mycelium for remediation of piroxicam.

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          Most cited references47

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          Lignin degradation: microorganisms, enzymes involved, genomes analysis and evolution

          Abstract Extensive research efforts have been dedicated to describing degradation of wood, which is a complex process; hence, microorganisms have evolved different enzymatic and non-enzymatic strategies to utilize this plentiful plant material. This review describes a number of fungal and bacterial organisms which have developed both competitive and mutualistic strategies for the decomposition of wood and to thrive in different ecological niches. Through the analysis of the enzymatic machinery engaged in wood degradation, it was possible to elucidate different strategies of wood decomposition which often depend on ecological niches inhabited by given organism. Moreover, a detailed description of low molecular weight compounds is presented, which gives these organisms not only an advantage in wood degradation processes, but seems rather to be a new evolutionatory alternative to enzymatic combustion. Through analysis of genomics and secretomic data, it was possible to underline the probable importance of certain wood-degrading enzymes produced by different fungal organisms, potentially giving them advantage in their ecological niches. The paper highlights different fungal strategies of wood degradation, which possibly correlates to the number of genes coding for secretory enzymes. Furthermore, investigation of the evolution of wood-degrading organisms has been described.
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            Toxic effects of the non-steroidal anti-inflammatory drug diclofenac. Part I: histopathological alterations and bioaccumulation in rainbow trout.

            Human and veterinary pharmaceuticals have been shown to occur in considerably high amounts in sewage treatment plant (STP) effluents and surface waters. The non-steroidal inflammatory drug diclofenac represents one of the most commonly detected compounds. Information concerning possible ecotoxicological risks of the substance are rather scarce. So far there are no data available on its possible effects in fish after prolonged exposure. In order to evaluate sublethal toxic effects of diclofenac in fish, rainbow trout (Oncorhynchus mykiss) exposed to diclofenac concentrations ranging from 1 microg/L to 500 microg/L over a 28 day period were investigated by histopathological methods. In addition, diclofenac residues in various organs were analyzed by means of gas chromatography/mass spectrometry (GC/MS). The histopathological examinations of diclofenac-exposed fish revealed alterations of the kidney such as an hyaline droplet degeneration of the tubular epithelial cells and the occurrence of an interstitial nephritis. In the gills, the predominant finding consisted in a necrosis of pillar cells leading to damage of the capillary wall within the secondary lamellae. The lowest observed effect concentration (LOEC) at which both renal lesions and alterations of the gills occurred was 5 microg/L. In contrast, the light microscopical examination of the liver, the gastro-intestinal tract, and the spleen did not reveal any histopathological alterations neither in diclofenac-exposed fish nor in solvent controls or control individuals. Chemical analysis showed a concentration-related accumulation of diclofenac in all organs examined. The highest amounts could be detected in the liver, followed by the kidney, the gills and the muscle tissue. Dependent on the diclofenac concentration used, the bioconcentration factors (BCF) were 12-2732 in the liver, 5-971 in the kidney, 3-763 in the gills, and 0.3-69 in the muscle respectively. From the present findings it can be assumed, that prolonged exposure in environmentally relevant concentrations of diclofenac leads to an impairment of the general health condition of fish.
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              Ability of white-rot fungi to remove selected pharmaceuticals and identification of degradation products of ibuprofen by Trametes versicolor.

              A screening using four white-rot fungi (Trametes versicolor, Irpex lacteus, Ganoderma lucidum and Phanerochaete chrysosporium) was performed on the degradation of 10 mg L(-1) of ibuprofen (IBU, anti-inflammatory), clofibric acid (CLOFI, lipid regulator) and carbamazepine (CARBA, antiepileptic/analgetic) after 7 d of incubation. Whereas IBU was extensively degraded by all the fungi tested, T. versicolor was the only strain able to degrade either CLOFI (approximately 91%) and CARBA (approximately 58%), although the latter was also degraded by G. lucidum (approximately 47%). In vitro experiments using manganese peroxidase and laccase-mediator system showed that extracellular fungal enzyme systems did not appear to play a role in the first step of degradation. However, our in vivo studies using the cytochrome P450 inhibitors 1-aminobenzotriazole and piperonyl butoxide suggested that the cytochrome P450 system may be involved in the first step of CLOFI and CARBA oxidation by T. versicolor. During the very early stages of IBU degradation by T. versicolor, two hydroxylated metabolites were detected: 1-hydroxy ibuprofen and 2-hydroxy ibuprofen. These byproducts were subsequently degraded by the fungus to 1,2-dihydroxy ibuprofen, that was not reported in biological systems to date. Furthermore, these results are of particular interest because CLOFI and CARBA are highly persistent in the aquatic environment and they pass unchanged or poorly transformed in wastewater treatment plants.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                21 February 2019
                2019
                : 10
                : 313
                Affiliations
                [1] 1Department of Pharmaceutical Botany, Faculty of Pharmacy, Jagiellonian University Medical College , Kraków, Poland
                [2] 2Department of Inorganic Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College , Kraków, Poland
                [3] 3Department of Medicinal Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College , Kraków, Poland
                [4] 4Department of Integrated Dentistry, Jagiellonian University Medical College , Kraków, Poland
                Author notes

                Edited by: Mariusz Cycoń, Medical University of Silesia, Poland

                Reviewed by: Robert Skibinski, Medical University of Lublin, Poland; Anna Gałązka, Institute of Soil Science and Plant Cultivation, Poland; Giovanna Cristina Varese, University of Turin, Italy; Santosh Kr. Karn, Sardar Bhagwan Singh Post Graduate Institute of Biomedical Science & Research, India

                *Correspondence: Monika Dąbrowska, mtylka@ 123456cm-uj.krakow.pl

                This article was submitted to Microbiotechnology, Ecotoxicology and Bioremediation, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2019.00313
                6393367
                30846979
                d6509ff8-156b-4a96-9e3b-185fd7cfe616
                Copyright © 2019 Muszyńska, Dąbrowska, Starek, Żmudzki, Lazur, Pytko-Polończyk and Opoka.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 10 December 2018
                : 05 February 2019
                Page count
                Figures: 2, Tables: 1, Equations: 0, References: 54, Pages: 9, Words: 0
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                bioremediation,edible mushroom,lentinula edodes,piroxicam,uplc/ms/ms analysis

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