Targeted isolation of antibiofilm compounds from halophytic endophyte Bacillus velezensis 7NPB-3B using LC-HR-MS-based metabolomics

Singh, Sanju and Nwagwu, Elizabeth and Young, Louise and Kumar, Pankaj and Shinde, Pramod B. and Edrada-Ebel, RuAngelie (2024) Targeted isolation of antibiofilm compounds from halophytic endophyte Bacillus velezensis 7NPB-3B using LC-HR-MS-based metabolomics. Microorganisms, 12 (2). 413. ISSN 2076-2607 (https://doi.org/10.3390/microorganisms12020413)

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Abstract

The discovery of new natural products has become more challenging because of the re-isolation of compounds and the lack of new sources. Microbes dwelling in extreme conditions of high salinity and temperature are huge prospects for interesting natural metabolites. In this study, the endophytic bacteria Bacillus velezensis 7NPB-3B isolated from the halophyte Salicornia brachiata was screened for its biofilm inhibition against methicillin-resistant Staphylococcus aureus (MRSA). The fractionation of the crude extract was guided by bioassay and LC-HRMS-based metabolomics using multivariate analysis. The 37 fractions obtained by high-throughput chromatography were dereplicated using an in-house MS-Excel macro coupled with the Dictionary of Natural Products database. Successive bioactivity-guided separation yielded one novel compound (1), a diketopiperazine (m/z 469.258 [M − H]−) with an attached saturated decanoic acid chain, and four known compounds (2–5). The compounds were identified based on 1D- and 2D-NMR and mass spectrometry. Compounds 1 and 5 exhibited excellent biofilm inhibition properties of >90% against the MRSA pathogen at minimum inhibition concentrations of 25 and 35 µg/mL, respectively. The investigation resulted in the isolation of a novel diketopiperazine from a bacterial endophyte of an untapped plant using an omics approach.

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