Metabolic characterisation of Magnetospirillum gryphiswaldense MSR-1 using LC-MS-based metabolite profiling

Salah Abdelrazig, Laudina Safo, Graham A. Rance, Michael W. Fay, Eirini Theodosiou, Paul D. Topham, Dong-hyun Kim, Alfred Fernández-castané*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Magnetosomes are nano-sized magnetic nanoparticles with exquisite properties that can be used in a wide range of healthcare and biotechnological applications. They are biosynthesised by magnetotactic bacteria (MTB), such as Magnetospirillum gryphiswaldense MSR-1 (Mgryph). However, magnetosome bioprocessing yields low quantities compared to chemical synthesis of magnetic nanoparticles. Therefore, an understanding of the intracellular metabolites and metabolic networks related to Mgryph growth and magnetosome formation are vital to unlock the potential of this organism to develop improved bioprocesses. In this work, we investigated the metabolism of Mgryph using untargeted metabolomics.
Liquid chromatography-mass spectrometry (LC-MS) was performed to profile spent medium samples of Mgryph cells grown under O2-limited (n ¼ 6) and O2-rich conditions (n = 6) corresponding to magnetosome- and non-magnetosome producing cells, respectively. Multivariate, univariate and pathway enrichment analyses were conducted to identify significantly altered metabolites and pathways.
Rigorous metabolite identification was carried out using authentic standards, the Mgryph-specific metabolite database and MS/MS mzCloud database. PCA and OPLS-DA showed clear separation and clustering of sample groups with cross-validation values of R2X ¼ 0.76, R2Y ¼ 0.99 and Q2 ¼ 0.98 in OPLS-DA. As a result, 50 metabolites linked to 45 metabolic pathways were found to be significantly altered in the tested conditions, including: glycine, serine and threonine; butanoate; alanine, aspartate and glutamate metabolism; aminoacyl-tRNA biosynthesis and; pyruvate and citric acid cycle (TCA) metabolisms. Our findings demonstrate the potential of LC-MS to characterise key metabolites in
Mgryph and will contribute to further understanding the metabolic mechanisms that affect Mgryph growth and magnetosome formation.
Original languageEnglish
Pages (from-to)32548-32560
Number of pages13
JournalRSC advances
Volume10
Issue number54
DOIs
Publication statusPublished - 2 Sep 2020

Bibliographical note

This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence.

Funding: This work was supported by the NanoPrime EPSRC grant EP/
R025282/1; Royal Society Research grant RGS\R1\191377; the
Aston Institute of Materials Research (AIMR) Seed-corn grant;
the Energy Research Accelerator (ERA) grant from Innovate UK
and the Green Chemicals Beacon of Excellence, University of
Nottingham.

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