Fluorine-containing substituents: metabolism of the α,α-difluoroethyl thioether motif

We report the metabolism of the recently introduced α,α-difluoroethyl thioether motif to explore further its potential as a substituent for bioactives discovery chemistry. Incubation of two aryl–SCF2CH3 ethers with the model yeast organism Cunninghamella elegans, indicates that the sulfur of the thioether is rapidly converted to the corresponding sulfoxide, and then significantly more slowly to the sulfone. When the substrate was (p-OMe)PhSCF2CH3, then the resultant (demethylated) phenol sulfoxide had an enantiomeric excess of 60%, and when the substrate was the β-substituted-SCF2CH3 naphthalene, then the enantiomeric excess of the resultant sulfoxide was 54%. There was no evidence of defluorination, unlike the corresponding oxygen ether (p-OMe)PhOCF2CH3, which was converted to the (demethylated) phenol acetate ester during C. elegans incubation. We conclude that the aryl–S–CF2CH3 motif is metabolised in a similar manner to aryl–SCF3, a motif that is being widely explored in discovery chemistry. It is however, significantly less lipophilic than aryl-SCF3 which may offer a practical advantage in tuning overall pharmacokinetic profiles of molecules in development.

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Biodegradation of Chloroxylenol by Cunninghamella elegans IM 1785/21GP and Trametes versicolor IM 373: Insight into Ecotoxicity and Metabolic Pathways

International Journal of Molecular Sciences ◽

10.3390/ijms22094360 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4360

Author(s):

Marta Nowak ◽

Katarzyna Zawadzka ◽

Janusz Szemraj ◽

Aleksandra Góralczyk-Bińkowska ◽

Katarzyna Lisowska

Keyword(s):

Trametes Versicolor ◽

Significant Rise ◽

Pcr Analysis ◽

Cunninghamella Elegans ◽

Oxidation Reactions ◽

Cytochrome P450 Enzymes ◽

C Elegans ◽

Real Time Quantitative Pcr ◽

Genes Expression ◽

By Products

Chloroxylenol (PCMX) is applied as a preservative and disinfectant in personal care products, currently recommended for use to inactivate the SARS-CoV-2 virus. Its intensive application leads to the release of PCMX into the environment, which can have a harmful impact on aquatic and soil biotas. The aim of this study was to assess the mechanism of chloroxylenol biodegradation by the fungal strains Cunninghamella elegans IM 1785/21GP and Trametes versicolor IM 373, and investigate the ecotoxicity of emerging by-products. The residues of PCMX and formed metabolites were analysed using GC-MS. The elimination of PCMX in the cultures of tested microorganisms was above 70%. Five fungal by-products were detected for the first time. Identified intermediates were performed by dechlorination, hydroxylation, and oxidation reactions catalysed by cytochrome P450 enzymes and laccase. A real-time quantitative PCR analysis confirmed an increase in CYP450 genes expression in C. elegans cells. In the case of T. versicolor, spectrophotometric measurement of the oxidation of 2,20-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) showed a significant rise in laccase activity during PCMX elimination. Furthermore, with the use of bioindicators from different ecosystems (Daphtoxkit F and Phytotoxkit), it was revealed that the biodegradation process of PCMX had a detoxifying nature.

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2C-B-Fly-NBOMe Metabolites in Rat Urine, Human Liver Microsomes and C. elegans: Confirmation with Synthesized Analytical Standards

Metabolites ◽

10.3390/metabo11110775 ◽

2021 ◽

Vol 11 (11) ◽

pp. 775

Author(s):

Jitka Nykodemová ◽

Anna Šuláková ◽

Petr Palivec ◽

Hedvika Češková ◽

Silvie Rimpelová ◽

...

Keyword(s):

Phase I ◽

Human Liver ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Diagnostic Tools ◽

Cunninghamella Elegans ◽

Rat Urine ◽

C Elegans ◽

Fragmentation Patterns

Compounds from the N-benzylphenethylamine (NBPEA) class of novel psychoactive substances are being increasingly utilized in neurobiological and clinical research, as diagnostic tools, or for recreational purposes. To understand the pharmacology, safety, or potential toxicity of these substances, elucidating their metabolic fate is therefore of the utmost interest. Several studies on NBPEA metabolism have emerged, but scarce information about substances with a tetrahydrobenzodifuran (“Fly”) moiety is available. Here, we investigated the metabolism of 2-(8-bromo-2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b’]difuran-4-yl)-N-(2-methoxybenzyl)ethan-1-amine (2C-B-Fly-NBOMe) in three different systems: isolated human liver microsomes, Cunninghamella elegans mycelium, and in rats in vivo. Phase I and II metabolites of 2C-B-Fly-NBOMe were first detected in an untargeted screening and identified by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Several hypothesized metabolites were then synthesized as reference standards; knowledge of their fragmentation patterns was utilized for confirmation or tentative identification of isomers. Altogether, thirty-five phase I and nine phase II 2C-B-Fly-NBOMe metabolites were detected. Major detected metabolic pathways were mono- and poly-hydroxylation, O-demethylation, oxidative debromination, and to a lesser extent also N-demethoxybenzylation, followed by glucuronidation and/or N-acetylation. Differences were observed for the three used media. The highest number of metabolites and at highest concentration were found in human liver microsomes. In vivo metabolites detected from rat urine included two poly-hydroxylated metabolites found only in this media. Mycelium matrix contained several dehydrogenated, N-oxygenated, and dibrominated metabolites.

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Transformation of chrysene and other polycyclic aromatic hydrocarbon mixtures by the fungus Cunninghamella elegans

Canadian Journal of Botany ◽

10.1139/b95-353 ◽

1995 ◽

Vol 73 (S1) ◽

pp. 1025-1033 ◽

Cited By ~ 20

Author(s):

Jairaj V. Pothuluri ◽

Allison Selby ◽

Frederick E. Evans ◽

James P. Freeman ◽

Carl E. Cerniglia

Keyword(s):

Polycyclic Aromatic Hydrocarbons ◽

Aromatic Hydrocarbon ◽

Aromatic Hydrocarbons ◽

High Performance ◽

Parent Compound ◽

Environmental Pollutants ◽

Health Concern ◽

Cunninghamella Elegans ◽

C Elegans ◽

Polycyclic Aromatic

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous and persistent environmental pollutants; some are mutagenic, toxic, and carcinogenic and remain a public health concern. We investigated the metabolism of mixtures of PAHs and a tetracyclic aromatic hydrocarbon, chrysene, by the filamentous fungus, Cunninghamella elegans ATCC 36112. Cunninghamella elegans metabolized a mixture of PAHs including the carcinogen benzo[a]pyrene, phenanthrene, fluoranthene, pyrene, and acenaphthene completely to hydroxylated intermediates within 24 h. The metabolites from the PAH mixtures were similar to those formed in earlier studies of individual PAH compounds. In separate experiments with chrysene, C. elegans metabolized about 45% of the [5,6,11,12-14C]chrysene added to cultures during 144 h incubation. The two major metabolites of chrysene were separated by reverse-phase high performance liquid chromatography and identified by ultraviolet–visible, mass spectral, and 1H-nuclear magnetic resonance techniques as sulfate conjugates of 2,8-dihydroxychrysene and 2-hydroxychrysene. The two major metabolites accounted for about 33% of the total metabolism. The formation of sulfate conjugates of phenolic chrysene metabolites and glucoside conjugates and hydroxylated products of PAH mixtures by C. elegans may be a detoxification step, because these types of products are generally less toxic than the parent compound. Key words: polycyclic aromatic hydrocarbons, PAH mixtures, chrysene, Cunninghamella elegans, biotransformation, oxidation.

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A Microbial Transformation Model for Simulating Mammal Metabolism of Artemisinin

Molecules ◽

10.3390/molecules24020315 ◽

2019 ◽

Vol 24 (2) ◽

pp. 315 ◽

Cited By ~ 3

Author(s):

Yue Ma ◽

Peng Sun ◽

Yifan Zhao ◽

Kun Wang ◽

Xiaoqiang Chang ◽

...

Keyword(s):

Chinese Herb ◽

Microbial Transformation ◽

Ultra Performance Liquid Chromatography ◽

The Body ◽

Transformation Model ◽

Cunninghamella Elegans ◽

C Elegans ◽

Traditional Chinese Herb

Artemisinin (ART) is a highly effective antimalarial agent isolated from the traditional Chinese herb Qinghao. Metabolism of ART and its derivatives in the body is one of the most pressing issues for pharmaceutical scientists. Herein, an efficient in vitro microorganism model for simulation of metabolism of ART in vivo was developed employing Cunninghamella elegans. Metabolites in the microbial transformation system and plasma of mice pre-administrated ART orally were analyzed by ultra-performance liquid chromatography (UPLC)-electrospray ionization (ESI)-quadrupole time-of-flight (Q-TOF)-mass spectrometry (MSE) combined with UNIFI software. Thirty-two metabolites were identified in vitro and 23 were identified in vivo. After comparison, 16 products were found to be common to both models including monohydroxylated ART, dihydroxylated ART, deoxyartemisinin, hydroxylated deoxyartemisinin, hydroxylated dihydroartemisinin (DHA), and hydroxylated deoxy-DHA. These results revealed that C. elegans CICC 40250 functioned as an appropriate model to mimic ART metabolism in vivo. Moreover, an overall description of metabolites of ART from C. elegans CICC 40250 has been provided. Notably, DHA was detected and identified as a metabolite of ART in mouse plasma for the first time.

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Regio- and stereo-selective metabolism of 4-methylbenz[a]anthracene by the fungus Cunninghamella elegans

Biochemical Journal ◽

10.1042/bj2160377 ◽

1983 ◽

Vol 216 (2) ◽

pp. 377-384 ◽

Cited By ~ 10

Author(s):

C E Cerniglia ◽

P P Fu ◽

S K Yang

Keyword(s):

Epoxide Hydrolase ◽

Microsomal Fraction ◽

Reversed Phase ◽

Cunninghamella Elegans ◽

Visible Absorption ◽

Spectral Techniques ◽

Methyl Group ◽

Mass Spectral ◽

C Elegans ◽

Enzyme Systems

Metabolism of 4-methylbenz[a]anthracene by the fungus Cunninghamella elegans was studied. C. elegans metabolized 4-methylbenz[a]anthracene primarily at the methyl group, this being followed by further metabolism at the 8,9- and 10,11-positions to form trans-8,9-dihydro-8,9-dihydroxy-4-hydroxymethylbenz[a]anthracene and trans-10,11-dihydro-10,11-dihydroxy-4-hydroxymethylbenz[a]anthracene. There was no detectable trans-dihydrodiol formed at the methyl-substituted double bond (3,4-positions) or at the ‘K’ region (5,6-positions). The metabolites were isolated by reversed-phase high-pressure liquid chromatography and characterized by the application of u.v.-visible-absorption-, 1H-n.m.r.- and mass-spectral techniques. The 4-hydroxymethylbenz[a]anthracene trans-8,9- and −10,11-dihydrodiols were optically active. Comparison of the c.d. spectra of the trans-dihydrodiols formed from 4-methylbenz[a]anthracene by C. elegans with those of the corresponding benz[a]anthracene trans-dihydrodiols formed by rat liver microsomal fraction indicated that the major enantiomers of the 4-hydroxymethylbenz[a]anthracene trans-8,9-dihydrodiol and trans- 10,11-dihydrodiol formed by C. elegans have S,S absolute stereochemistries, which are opposite to those of the predominantly 8R,9R- and 10R,11R-dihydrodiols formed by the microsomal fraction. Incubation of C. elegans with 4-methylbenz[a]anthracene under 18O2 and subsequent mass-spectral analysis of the metabolites indicated that hydroxylation of the methyl group and the formation of trans-dihydrodiols are catalysed by cytochrome P-450 mono-oxygenase and epoxide hydrolase enzyme systems. The results indicate that the fungal mono-oxygenase-epoxide hydrolase enzyme systems are highly stereo- and regio-selective in the metabolism of 4-methylbenz[a]anthracene.

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Mucorales isolados do solo de mineração de cobre e produção de amilase e inulinase

Acta Botanica Brasilica ◽

10.1590/s0102-33062006000300014 ◽

2006 ◽

Vol 20 (3) ◽

pp. 641-647 ◽

Cited By ~ 3

Author(s):

André Luiz Cabral Monteiro Azevedo Santiago ◽

Cristina Maria de Souza-Motta

Keyword(s):

Rhizopus Oryzae ◽

Cunninghamella Elegans ◽

Rhizopus Microsporus ◽

Syncephalastrum Racemosum ◽

C Elegans

A presença de metais pesados no solo proporciona impacto sobre os microrganismos, vegetação e os processos funcionais do ecossistema. Visando comprovar que Mucorales são afetados pela extração de cobre na Mineradora Caraíba, Jaguarari, BA, espécies de Mucorales foram isoladas de amostras de solo coletadas nas épocas seca e chuvosa. Os Mucorales foram identificados e caracterizados quanto à capacidade de degradar amido e inulina. Utilizando os métodos de diluição e placa de solo, foram obtidos 46 isolados de Mucorales pertencentes a sete diferentes espécies. Absidia blakesleeana Lendner, A. cylindrospora Hagem, A. hialospora (Saito) Lendn., Cunninghamella elegans Lendner, Rhizopus microsporus V. Thieghen, R. oryzae Went. & Prinsen Geerl. e Syncephalastrum racemosum (Cohn.) Schroet. foram isolados na época seca. A. blakesleeana e A. hialospora foram isoladas apenas na época chuvosa. Rhizopus oryzae apresentou maior número de isolados (30,43%), seguido por A. blakesleeana (26,09%) e C. elegans (21,74%). O gênero que apresentou maior número de espécies foi Absidia seguido por Rhizopus. Entre as áreas, houve diferenças significativas no número de isolados de Mucorales, comprovando os efeitos negativos do impacto ambiental na abundância de Mucorales nos solos impactados. Todas as espécies degradaram o amido e apenas C. elegans degradou também inulina. Culturas de mesma espécie isoladas de áreas diferentes apresentaram o mesmo comportamento quanto à degradação de amido e inulina.

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Substrate Specificity and Enantioselectivity of 4-Hydroxyacetophenone Monooxygenase

Applied and Environmental Microbiology ◽

10.1128/aem.69.1.419-426.2003 ◽

2003 ◽

Vol 69 (1) ◽

pp. 419-426 ◽

Cited By ~ 85

Author(s):

Nanne M. Kamerbeek ◽

Arjen J. J. Olsthoorn ◽

Marco W. Fraaije ◽

Dick B. Janssen

Keyword(s):

Carbonyl Compounds ◽

Kinetic Resolution ◽

Enantiomeric Excess ◽

The Other ◽

Biotechnological Applications ◽

Complete Conversion ◽

Aromatic Ketones ◽

Acetate Ester ◽

Enantiomeric Ratio ◽

Villiger Oxidation

ABSTRACT The 4-hydroxyacetophenone monooxygenase (HAPMO) from Pseudomonas fluorescens ACB catalyzes NADPH- and oxygen-dependent Baeyer-Villiger oxidation of 4-hydroxyacetophenone to the corresponding acetate ester. Using the purified enzyme from recombinant Escherichia coli, we found that a broad range of carbonylic compounds that are structurally more or less similar to 4-hydroxyacetophenone are also substrates for this flavin-containing monooxygenase. On the other hand, several carbonyl compounds that are substrates for other Baeyer-Villiger monooxygenases (BVMOs) are not converted by HAPMO. In addition to performing Baeyer-Villiger reactions with aromatic ketones and aldehydes, the enzyme was also able to catalyze sulfoxidation reactions by using aromatic sulfides. Furthermore, several heterocyclic and aliphatic carbonyl compounds were also readily converted by this BVMO. To probe the enantioselectivity of HAPMO, the conversion of bicyclohept-2-en-6-one and two aryl alkyl sulfides was studied. The monooxygenase preferably converted (1R,5S)-bicyclohept-2-en-6-one, with an enantiomeric ratio (E) of 20, thus enabling kinetic resolution to obtain the (1S,5R) enantiomer. Complete conversion of both enantiomers resulted in the accumulation of two regioisomeric lactones with moderate enantiomeric excess (ee) for the two lactones obtained [77% ee for (1S,5R)-2 and 34% ee for (1R,5S)-3]. Using methyl 4-tolyl sulfide and methylphenyl sulfide, we found that HAPMO is efficient and highly selective in the asymmetric formation of the corresponding (S)-sulfoxides (ee > 99%). The biocatalytic properties of HAPMO described here show the potential of this enzyme for biotechnological applications.

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25CN-NBOMe Metabolites in Rat Urine, Human Liver Microsomes and C.elegans—Structure Determination and Synthesis of the Most Abundant Metabolites

Metabolites ◽

10.3390/metabo11040212 ◽

2021 ◽

Vol 11 (4) ◽

pp. 212

Author(s):

Anna Šuláková ◽

Jitka Nykodemová ◽

Petr Palivec ◽

Radek Jurok ◽

Silvie Rimpelová ◽

...

Keyword(s):

Human Liver ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Cunninghamella Elegans ◽

Rat Urine ◽

C Elegans ◽

Liquid Chromatography Tandem Mass ◽

Phase Ii Metabolites

N-Benzylphenethylamines are novel psychedelic substances increasingly used for research, diagnostic, or recreational purposes. To date, only a few metabolism studies have been conducted for N-2-methoxybenzylated compounds (NBOMes). Thus, the available 2,5-dimethoxy-4-(2-((2-methoxybenzyl)amino)ethyl)benzonitrile (25CN-NBOMe) metabolism data are limited. Herein, we investigated the metabolic profile of 25CN-NBOMe in vivo in rats and in vitro in Cunninghamella elegans (C. elegans) mycelium and human liver microsomes. Phase I and phase II metabolites were first detected in an untargeted screening, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification of the most abundant metabolites by comparison with in-house synthesized reference materials. The major metabolic pathways described within this study (mono- and bis-O-demethylation, hydroxylation at different positions, and combinations thereof, followed by the glucuronidation, sulfation, and/or N-acetylation of primary metabolites) generally correspond to the results of previously reported metabolism of several other NBOMes. The cyano functional group was either hydrolyzed to the respective amide or carboxylic acid or remained untouched. Differences between species should be taken into account in studies of the metabolism of novel substances.

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Enhanced Biotransformation of Fluoranthene by Intertidally Derived Cunninghamella elegans under Biofilm-Based and Niche-Mimicking Conditions

Applied and Environmental Microbiology ◽

10.1128/aem.02129-13 ◽

2013 ◽

Vol 79 (24) ◽

pp. 7922-7930 ◽

Cited By ~ 10

Author(s):

Sayani Mitra ◽

Arnab Pramanik ◽

Srijoni Banerjee ◽

Saubhik Haldar ◽

Ratan Gachhui ◽

...

Keyword(s):

Gene Expression ◽

Cytochrome P450 ◽

Glass Surface ◽

Biofilm Formation ◽

Cunninghamella Elegans ◽

Biofilm Growth ◽

Cytochrome P450 Gene ◽

Content Type ◽

P450 Gene ◽

C Elegans

ABSTRACTThe aims of the investigation were to ascertain if surface attachment ofCunninghamella elegansand niche intertidal conditions provided in a bioreactor influenced biotransformation of fluoranthene byC. elegans. A newly designed polymethylmethacrylate (PMMA) conico-cylindrical flask (CCF) holding eight equidistantly spaced rectangular strips mounted radially on a circular disc allowed comparison of fluoranthene biotransformation between CCFs with a hydrophobic surface (PMMA-CCF) and a hydrophilic glass surface (GS-CCF) and a 500-ml Erlenmeyer flask (EF). Fluoranthene biotransformation was higher by 22-fold, biofilm growth was higher by 3-fold, and cytochrome P450 gene expression was higher by 2.1-fold whenC. eleganswas cultivated with 2% inoculum as biofilm culture in PMMA-CCF compared to planktonic culture in EF. Biotransformation was enhanced by 7-fold with 10% inoculum. The temporal pattern of biofilm progression based on three-channel fluorescence detection by confocal laser scanning microscopy demonstrated well-developed, stable biofilm with greater colocalization of fluoranthene within extracellular polymeric substances and filaments of the biofilm grown on PMMA in contrast to a glass surface. A bioreactor with discs rotating at 2 revolutions per day affording 6-hourly emersion and immersion mimicked the niche intertidal habitat ofC. elegansand supported biofilm formation and transformation of fluoranthene. The amount of transformed metabolite was 3.5-fold, biofilm growth was 3-fold, and cytochrome P450 gene expression was 1.9-fold higher in the process mimicking the intertidal conditions than in a submerged process without disc rotation. In the CCF and reactor, where biofilm formation was comparatively greater, higher concentration of exopolysaccharides allowed increased mobilization of fluoranthene within the biofilm with consequential higher gene expression leading to enhanced volumetric productivity.

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BIORREMOÇÃO DE FENANTRENO POR BIOMASSA VIVA E INATIVADA CUNNINGHAMELLA ELEGANS UCP0542

e-xacta ◽

10.18674/exacta.v6i1.839 ◽

2013 ◽

Vol 6 (1) ◽

pp. 1 ◽

Cited By ~ 1

Author(s):

Marta Cristina Freitas Silva ◽

Adriana Almeida Antunes ◽

Clarissa Isabel Matos Lins ◽

Antonio Helder Parente ◽

Sônia Valéria Pereira ◽

...

Keyword(s):

Polycyclic Aromatic Hydrocarbons ◽

Aromatic Hydrocarbons ◽

Cunninghamella Elegans ◽

Biotechnological Potential ◽

Independent Variables ◽

C Elegans ◽

Live Biomass ◽

Polycyclic Aromatic ◽

Full Factorial ◽

Inactivated Biomass

Realizou-se estudos avaliando a ação da biomassa viva e inativada de Cunninghamella elegans crescida, visando à remoção do hidrocarboneto aromático policíclico fenantreno. Monitorou-se o processo de remoção do fenantreno a cada 24 h por espectrofotometria, de acordo com um planejamento fatorial completo de 22 e, como variáveis independentes 0,1 e 0,2 mM de fenantreno, na presença de 0,6, 3,3 e 6% de glicose e variável resposta o conteúdo de fenantreno removido. Os resultados obtidos indicaram que tanto a biomassa inativada como viva removeram, respectivamente, 90,0% e 82,0% a concentração de 0,2mM de fenantreno na presença da menor concentração de glicose (0,6%). A biomassa inativada de C. elegans demonstrou ser um sorbente com elevado potencial biotecnológico para a remediação de áreas poluídas com hidrocarbonetos aromáticos policíclicos.AbstractStudies were carried out evaluating the action of the life and inactivated biomass of Cunninghamella elegans grown to removal polycyclic aromatic hydrocarbon phenanthrene. The removal process was monitored each 24h by spectrophotometry, de acordo com according to full factorial design 2 2 , and independent variables the phenanthrene concentrations [0.1 and 0.2 mM], and the content of phenanthrene removed as variable response.The results showed that both inactivated and live biomass from C. elegans removed 85.0% and 90.0%, respectively, of phenantrene of 0.2 mM on glucose (0.6) concentration. The inactivated biomass from C. elegans shows a sorbent with high biotechnological potential for bioremediation of polluted area with polycyclic aromatic hydrocarbons.

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