Metabolic profiles and fingerprints for the investigation of the influence of nitisinone on the metabolism of the yeast Saccharomyces cerevisiae

Abstract

Nitisinone (2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione, NTBC) is considered a potentially effective drug for the treatment of various metabolic diseases associated with disorders of l-tyrosine metabolism however, side-effects impede its widespread use. This work aimed to broaden the knowledge of the influence of NTBC and its metabolites 2-amino-4-(trifluoromethyl)benzoic acid (ATFA), 2-nitro-4-(trifluoromethyl)benzoic acid (NTFA), and cyclohexane-1,3-dione (CHD) on the catabolism of l-tyrosine and other endogenous compounds in Saccharomyces cerevisiae. Based on a targeted analysis performed by LC–ESI–MS/MS, based on multiple reaction monitoring, it was found that the dissipation kinetics of the parent compound and its metabolites are compatible with a first-order reaction mechanism. Moreover, it has been proven that formed NTBC metabolites, such as CHD, cause a decrease in l-tyrosine, l-tryptophan, and l-phenylalanine concentrations by about 34%, 59% and 51%, respectively, compared to the untreated model organism. The overall changes in the metabolism of yeast exposed to NTBC or its derivatives were evaluated by non-targeted analysis via LC–ESI–MS/MS in the ion trap scanning mode. Based on principal components analysis, a statistically significant similarity between metabolic responses of yeast treated with ATFA or NTFA was observed. These findings facilitate further studies investigating the influence of NTBC on the human body and the mechanism of its action.

Cladosporium sphaerospermum extract inhibits quorum sensing associated virulence factors of Serratia marcescens

“…It has been reported to have numerous biological activities, including anti-inflammatory, antitumor, and antibacterial properties against L. hongkongensis , M. luteus and Ruegeria sp [ 29 , 30 ]. The compound with a Rt of 11.5 min was 3-phenyllactic acid which exhibited QSI and anti-biofilm activity against P. aeruginosa [ 21 , 31 ]. Anthranilic acid with a Rt of 10.0 min, could inhibit hypha formation in S. scitamineum [ 32 , 33 ].…”

Metabolic profiling to evaluate the impact of amantadine and rimantadine on the secondary metabolism of a model organism

“…Subsequently, each sample was filtered through a nylon filter with a pore diameter of 0.22 μm, and 1 mL of the clear extract was transferred to a vial for chromatographic analysis. Detailed information on the optimization of extraction conditions is available in our article 31 . The procedure was repeated for all sample types in triplicate.…”

“…The tuning and optimization of the compound-dependent parameters, including the declustering potential (DP), collision energy (CE), entrance potential (EP), and collision exit potential (CXP), were performed through the direct infusion of a standard solution into the ion source using a Harvard syringe pump. The pairs of monitoring ions for AMT and RMT, as well as compound-dependent MS parameters, are shown in Table 3SM, while selected endogenous compounds were presented in our previous article 31 . The protonated molecular ions [M + H] + of AMT and RMT at the optimized conditions of QTRAP MS were at m/z 152.108 and 180.644, respectively, for details, see Supplementary Material, Figure 1SM .…”

“…They concluded that, despite some doubts, Saccharomyces cerevisiae is a suitable model organism for higher organisms 26 , 27 . To date, Saccharomyces cerevisiae as a model organism was employed in the study of various respiratory dysfunctions 28 , aging processes 29 , and neurodegenerative 30 and metabolic 31 disorders.…”

Metabolic profiling to evaluate the impact of amantadine and rimantadine on the secondary metabolism of a model organism

“…Subsequently, each sample was ltered through a nylon lter with a pore diameter of 0.22 µm, and 1 mL of the clear extract was transferred to a vial for chromatographic analysis. Detailed information on the optimization of extraction conditions is available in our article 31 . The procedure was repeated for all sample types in triplicate.…”