Microbial transformation of ambrisentan to its glycosides by <i>Cunninghamella elegans</i>

Sponchiado, Rafaela Martins; Sorrentino, Julia Medeiros; Olegário, Natália; Oliveira, Sendy Sales; Cordenonsi, Leticia Malgarim; Silveira, Gustavo Pozza; Mendez, Andreas Sebastian Loureiro; Steppe, Martin; Garcia, Cássia Virgínia

doi:10.1002/bmc.4496

Cited by 7 publications

(5 citation statements)

References 21 publications

(27 reference statements)

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“…The activated bacterial cells were incubated at 28 °C for about seven days until the mycelium grew vigorously and spores were abundant. The spores on the plate were washed into a sterile triangle bottle with 5 mL of sterile water to make spore fluid [ 53 ]. The number of spores in the spore fluid was counted by hemocytometry to determine the inoculation amount.…”

Section: Methodsmentioning

confidence: 99%

The Study of a Novel Paeoniflorin-Converting Enzyme from Cunninghamella blakesleeana

Pei

Cao

et al. 2023

Molecules

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Paeoniflorin is a glycoside compound found in Paeonia lactiflora Pall that is used in traditional herbal medicine and shows various protective effects on the cardio-cerebral vascular system. It has been reported that the pharmacological effects of paeoniflorin might be generated by its metabolites. However, the bioavailability of paeoniflorin by oral administration is low, which greatly limits its clinical application. In this paper, a paeoniflorin-converting enzyme gene (G6046, GenBank accession numbers: OP856858) from Cunninghamella blakesleeana (AS 3.970) was identified by comparative analysis between MS analysis and transcriptomics. The expression, purification, enzyme activity, and structure of the conversion products produced by this paeoniflorin-converting enzyme were studied. The optimal conditions for the enzymatic activity were found to be pH 9, 45 °C, resulting in a specific enzyme activity of 14.56 U/mg. The products were separated and purified by high-performance counter-current chromatography (HPCCC). Two main components were isolated and identified, 2-amino-2-p-hydroxymethyl-methyl alcohol-benzoate (tirs-benzoate) and 1-benzoyloxy-2,3-propanediol (1-benzoyloxypropane-2,3-diol), via UPLC-Q-TOF-MS and NMR. Additionally, paeoniflorin demonstrated the ability to metabolize into benzoic acid via G6046 enzyme, which might exert antidepressant effects through the blood–brain barrier into the brain.

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Section: Methodsmentioning

confidence: 99%

The Study of a Novel Paeoniflorin-Converting Enzyme from Cunninghamella blakesleeana

Pei

Cao

et al. 2023

Molecules

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“…The work here presented as a part of our ongoing efforts focused on the development and optimization of biotransformation of drug studies and e validation of the analytical method. In previous studies, several parameters related to the culture medium were modified during the process of biotransformation of drugs, such as the amount of oxygen, pH, temperature, and speed of culture medium shacking (21,22). According to those parameters, modifications were done to optimize de biotransformation process.…”

Section: Fungal Biotransformation Studymentioning

confidence: 99%

Biotransformation of Metronidazole by Cunninghamella Elegans Atcc 9245

et al. 2019

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Drug biotransformation studies appear as an alternative to pharmacological studies of metabolites, development of new drug candidates with reduced investment as well as the most efficient production of chemical structures involves and drug quality control studies. A wide range of reactions in biotransformations process is catalyzed by microorganisms. Fungi can be considered as a promising source of new biotransformation reactions. The aim of this study was to evaluate the capacity of metronidazole biotransformation through the filamentous fungus Cunninghamella elegans ATCC 9245. The monitoring of metabolite formation was performed by high-performance liquid chromatography (HPLC) coupled to ultraviolet (UV) spectrophotometry. The results of the biotransformation of metronidazole showed drug consumption in culture and the formation of four new chromatographic peaks of chemical structures not elucidated. The method showed it became linear over 10-70 μg/mL (r = 0.999953). Accuracy, precision and stability studies agree with international guidelines. Results are consistent in accordance with the principles of green chemistry as the experimental conditions had a low environmental impact, and few solvents use.

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“…Generally, supplementary methods can be used to obtain a sufficient sample amount of metabolites, including chemical methods [11] and small experimental animal models [15], microsomal preparations [16,17], enzyme-catalyzed reactions [18,19], microbial transformation [11,20] and so on. Compared with other methods, microbial transformation is more convenient and economical, especially with the advantages of in vitro large-scale preparation [20][21][22][23][24]. In particular, the filamentous zygote fungus C. elegans has been shown to possess a human-like cytochrome P450 monooxygenase system, including the CYP509A1 isoenzyme that is similar to the CYP51 family, thus producing similar metabolic profiles to mammalian animals [25,26].Sufficient samples for structural characterization based on NMR techniques as well as the evaluation of bioactivity and toxicity can thus be acquired.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive Metabolic Profiling of Euphorbiasteroid in Rats by Integrating UPLC-Q/TOF-MS and NMR as Well as Microbial Biotransformation

Xiao¹,

Wei

Xin

et al. 2022

Metabolites

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Euphorbiasteroid, a lathyrane-type diterpene from Euphorbiae semen (the seeds of Euphorbia lathyris L.), has been shown to have a variety of pharmacological effects such as anti-tumor and anti-obesity. This study aims to investigate the metabolic profiles of euphorbiasteroid in rats and rat liver microsomes (RLMs) and Cunninghamella elegans bio-110930 by integrating ultra-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-Q/TOF-MS), UNIFI software, and NMR techniques. A total of 31 metabolites were identified in rats. Twelve metabolites (M1–M5, M8, M12–M13, M16, M24–M25, and M29) were matched to the metabolites obtained by RLMs incubation and the microbial transformation of C. elegans bio-110930 and their structures were exactly determined through analysis of NMR spectroscopic data. In addition, the metabolic pathways of euphorbiasteroid were then clarified, mainly including hydroxylation, hydrolysis, oxygenation, sulfonation, and glycosylation. Finally, three metabolites, M3 (20-hydroxyl euphorbiasteroid), M24 (epoxylathyrol) and M25 (15-deacetyl euphorbiasteroid), showed significant cytotoxicity against four human cell lines with IC50 values from 3.60 μM to 40.74 μM. This is the first systematic investigation into the in vivo metabolic pathways of euphorbiasteroid and the cytotoxicity of its metabolites, which will be beneficial for better predicting the metabolism profile of euphorbiasteroid in humans and understanding its possible toxic material basis.

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Microbial transformation of ambrisentan to its glycosides by Cunninghamella elegans

Cited by 7 publications

References 21 publications

The Study of a Novel Paeoniflorin-Converting Enzyme from Cunninghamella blakesleeana

The Study of a Novel Paeoniflorin-Converting Enzyme from Cunninghamella blakesleeana

Biotransformation of Metronidazole by Cunninghamella Elegans Atcc 9245

Comprehensive Metabolic Profiling of Euphorbiasteroid in Rats by Integrating UPLC-Q/TOF-MS and NMR as Well as Microbial Biotransformation

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