LC-QTOF-MS/MS metabolite profiling data from ethyl acetate extracts of Penicillium brevicompactum in modified glucose nutrient medium

Published: 5 April 2024| Version 3 | DOI: 10.17632/ksgsr5mfg4.3
Contributors:
Lukhanyo Mekuto,

Description

This data represents the metabolites which are produced by Penicillium brevicompactum in modified glucose medium as determined by LC-QTOF-MS/MS.

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Experimental Design, materials and methods Isolation of microorganism P. brevicompactum was isolated from an unidentified brown mushroom in a forest dominated by Celtis Africana in Cape Town, South Africa [1]. Microorganism biotransformation 1 L of modified glucose-nutrient (MGN) medium was prepared where the composition of MGN contained 5 g/L of peptone, 5 g/L dipotassium phosphate, 5 g/L yeast extract and 5 g/L sodium chloride. The mixture was sterilized at 121 ℃ for 90 minutes. Once the medium was cooled to 50 °C, 20 g/L of glucose was added to the medium. The spores of the newly grown Penicillium brevicompactum were inoculated in MGN medium to the 1 L flask. The amalgam was incubated at 30 °C for 9 days at stagnant conditions. After the incubation period, the mixture was transferred into falcon tubes and centrifuged at 4000 rpm for 20 minutes at 4 °C to remove the cell debris. Equal volumes of the supernatant and ethyl acetate were then mixed in a separating funnel and allowed to stand for approximately 30 minutes – 1 hour. The solvent phase was then concentrated using a rotary evaporator set at 70 °C and the resulting mixture was dried overnight in an oven to remove any additional solvent still present. The final blends were analysed using LC-QTOF-MS. This was done to detect the metabolites that this organism produced. Metabolite profiling using LC-QTOF-MS The analysis and detection of the metabolites were detected using the LC-QTOF system with a Dionex UltiMate 3000 UHPLC (Thermo Scientific, Darmstadt, Germany) coupled to a Compact™ QTOF (Bruker Daltonics, Bremen, Germany) [2]. 5 µl of the prepared sample was injected in the system for chromatographic separation of analytes in reverse phase ultra-high-performance liquid chromatography (RP-UHPLC) through a Raptor ARC-18 column (Restek, Bellefonte, USA). The mobile phase was composed of solvent A (A) consisting of 0.1 % formic acid in H2O (v/v) and solvent B (B) consisting of 0.1 % formic acid in acetonitrile (v/v). The operation and control of the instrument including data acquisition was done using HyStar software version 2.10 (Thermo Scientific, Darmstadt, Germany) while the spectral data processing was performed using the Bruker Compass DataAnalysis software version 4.3 (Bruker Daltonics, Bremen, Germany). MetFrag Handler version 2.1 software (GitHub, California, USA) was used to characterize the resulting fragment spectra by linking to three compound databases, namely PubChem, ChemSpider and KEGG [3]. Additional databases that were used included METLIN (Scripps Research, California, USA), KNapSAcK (Kanaya Laboratory, Japan), and Fungalmet (LINFA Project, Porto Salvo Vibo Valentia, Italy) a database primarily used for fungal metabolites. Control samples were analysed in the same conditions as above using the utilized growth medium such that the impurities are filtered during data interpretation.

Institutions

University of Johannesburg - Doornfontein Campus

Categories

Biochemistry, Metabolite, Biocatalysis, Penicillium, Liquid Chromatography Tandem Mass Spectrometry

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