Moeder, Monika* Dotaz Zobrazit nápovědu
A group of white rot fungi (Irpex lacteus, Panus tigrinus, Dichomitus squalens, Trametes versicolor and Pleurotus ostreatus) was investigated for the biodegradation of norfloxacin (NOR), ofloxacin (OF) and ciprofloxacin (CIP). The selected fluoroquinolones were readily degraded almost completely by I. lacteus and T. versicolor within 10 and 14 d of incubation in liquid medium, respectively. The biodegradation products were identified by liquid chromatography-mass spectrometry. The analyses indicated that the fungi use similar mechanisms to degrade structurally related antibiotics. The piperazine ring of the molecules is preferably attacked via either substitution or/and decomposition. In addition to the degradation efficiency, attention was devoted to the residual antibiotic activities estimated using Gram-positive and Gram-negative bacteria. Only I. lacteus was able to remove the antibiotic activity during the course of the degradation of NOR and OF. The product-effect correlations evaluated by Principal Component Analysis (PCA) enabled elucidation of the participation of the individual metabolites in the residual antibacterial activity. Most of the metabolites correlated with the antibacterial activity, explaining the rather high residual activity remaining after the biodegradation. PCA of ligninolytic enzyme activities indicated that manganese peroxidase might participate in the degradation.
- MeSH
- antibakteriální látky metabolismus MeSH
- biodegradace MeSH
- biotransformace * MeSH
- fluorochinolony metabolismus MeSH
- hmotnostní spektrometrie MeSH
- norfloxacin MeSH
- ofloxacin MeSH
- peroxidasy MeSH
- Pleurotus metabolismus MeSH
- Polyporales metabolismus MeSH
- Trametes metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The white rot fungus Pleurotus ostreatus is able to completely remove the synthetic hormone 17α-ethinylestradiol (EE2, 200 μg in 20 mL) from a liquid complex or mineral medium in 3 or 14 days, respectively. Its efficiency has also been documented in the removal of estrogenic activity that correlated with the EE2 degradation. A set of in vitro experiments using various cellular and enzyme fractions has been performed and the results showed that EE2 was degraded by isolated laccase (about 90% within 24 h). The degradation was also tested with concentrated extracellular liquid where degradation reached 50% mainly due to the laccase activity; however, after a supplementation with H₂O₂ and Mn²⁺, residual manganese-dependent peroxidase activities (40 times lower than Lac) raised the degradation to 100%. Moreover, the intracellular fraction and also laccase-like activity associated with fungal mycelium were found to be efficient in the degradation too. Isolated microsomal proteins appeared to also be involved in the process. The degradation was completely suppressed in the presence of cytochrome P-450 inhibitors, piperonylbutoxide and carbon monoxide, indicating a role of this monooxygenase in the degradation process. Attention was also paid to monitoring of changes in the estrogenic activity during these particular in vitro experiments when mainly degradations related to ligninolytic enzymes were found to decrease the estrogenic activity with EE2 removal proportionally. Several novel metabolites of EE2 were detected using different chromatographic method with mass spectrometric techniques (LC-MS, GC-MS) including also [¹³C]-labeled substrates. The results document the involvement of various different simultaneous mechanisms in the EE2 degradation by P. ostreatus by both the ligninolytic system and the eukaryotic machinery of cytochromes P-450.
- MeSH
- biodegradace MeSH
- ethinylestradiol chemie izolace a purifikace MeSH
- extracelulární prostor metabolismus MeSH
- intracelulární prostor metabolismus MeSH
- kultivační média MeSH
- lignin metabolismus MeSH
- metabolom MeSH
- mycelium metabolismus MeSH
- Pleurotus metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Flumequine, a fluoroquinolone antibiotic, is applied preferably in veterinary medicine, for stock breeding and treatment of aquacultures. Formation of drug resistance is a matter of general concern when antibiotics such as flumquine occur in the environment. Thus, biodegradation of flumequine in solution was investigated using five different ligninolytic fungi. Irpex lacteus, Dichomitus squalens, and Trametes versicolor proved most efficient and transformed more than 90% of flumequine within 6 or even 3 days. Panus tigrinus and Pleurotus ostreatus required up to 14 days to remove >90% of flumequine. Analyses of the metabolites by liquid chromatography-mass spectrometry suggest different transformation pathways for the different fungal strains. Structure proposals were elaborated for 8 metabolites. 7-Hydroxy-flumequine and flumequine ethyl ester were identified as common metabolites produced by all ligninolytic fungi. The largest variety of metabolites was formed by D. squalens. Residual antibacterial activity of the metabolite mixtures was tested using gram-positive and gram-negative bacteria. While for the less efficient P. tigrinus and P. ostreatus cultures the antibacterial activities corresponded to the residual concentrations of flumequine, a remarkable antibacterial activity remained in the D. squalens cultures although flumequine was transformed to more than 90%. Obviously, antibacterially active transformation products were formed by this fungal strain.
- MeSH
- analýza hlavních komponent MeSH
- antibakteriální látky chemie metabolismus farmakologie MeSH
- Bacteria účinky léků MeSH
- biodegradace účinky léků MeSH
- biotransformace účinky léků MeSH
- fluorochinolony chemie metabolismus farmakologie MeSH
- hmotnostní spektrometrie MeSH
- houby metabolismus MeSH
- kultivační média chemie MeSH
- lignin metabolismus MeSH
- mikrobiální testy citlivosti MeSH
- Pleurotus metabolismus MeSH
- Trametes metabolismus MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Due to their enhanced reactivity, metal and metal-oxide nanoscale zero-valent iron (nZVI) nanomaterials have been introduced into remediation practice. To ensure that environmental applications of nanomaterials are safe, their possible toxic effects should be described. However, there is still a lack of suitable toxicity tests that address the specific mode of action of nanoparticles, especially for nZVI. This contribution presents a novel approach for monitoring one of the most discussed adverse effects of nanoparticles, i.e., oxidative stress (OS). We optimized and developed an assay based on headspace-SPME-GC-MS analysis that enables the direct determination of volatile oxidative damage products (aldehydes) of lipids and proteins in microbial cultures after exposure to commercial types of nZVI. The method employs PDMS/DVB SPME fibers and pentafluorobenzyl derivatization, and the protocol was successfully tested using representatives of bacteria, fungi, and algae. Six aldehydes, namely, formaldehyde, acrolein, methional, benzaldehyde, glyoxal, and methylglyoxal, were detected in the cultures, and all of them exhibited dose-dependent sigmoidal responses. The presence of methional, which was detected in all cultures except those including an algal strain, documents that nZVI also caused oxidative damage to proteins in addition to lipids. The most sensitive toward nZVI exposure in terms of aldehyde production was the yeast strain Saccharomyces cerevisiae, which had an EC50 value of 0.08 g/L nZVI. To the best of our knowledge, this paper is the first to document the production of aldehydes resulting from lipids and proteins as a result of OS in microorganisms from different kingdoms after exposure to iron nanoparticles.
