title: Metabolomic tools for secondary metabolite discovery from marine microbial symbionts.
abstract: Marine invertebrate-associated symbiotic bacteria produce a plethora of novel secondary metabolites which may be structurally unique with interesting pharmacological properties. Selection of strains usually relies on literature searching, genetic screening and bioactivity results, often without considering the chemical novelty and abundance of secondary metabolites being produced by the microorganism until the time-consuming bioassay-guided isolation stages. To fast track the selection process, metabolomic tools were used to aid strain selection by investigating differences in the chemical profiles of 77 bacterial extracts isolated from cold water marine invertebrates from Orkney, Scotland using liquid chromatography-high resolution mass spectrometry (LC-HRMS) and nuclear magnetic resonance (NMR) spectroscopy. Following mass spectrometric analysis and dereplication using an Excel macro developed in-house, principal component analysis (PCA) was employed to differentiate the bacterial strains based on their chemical profiles. NMR 1H and correlation spectroscopy (COSY) were also employed to obtain a chemical fingerprint of each bacterial strain and to confirm the presence of functional groups and spin systems. These results were then combined with taxonomic identification and bioassay screening data to identify three bacterial strains, namely Bacillus sp. 4117, Rhodococcus sp. ZS402 and Vibrio splendidus strain LGP32, to prioritize for scale-up based on their chemically interesting secondary metabolomes, established through dereplication and interesting bioactivities, determined from bioassay screening.
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK. lynsey.macintyre@strath.ac.uk.
authors.forename: Lynsey
authors.initials: L
authors.lastname: Macintyre
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK. tong.zhang.101@strath.ac.uk.
authors.forename: Tong
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK. christina.viegelmann@strath.ac.uk.
authors.forename: Christina
authors.initials: C
authors.lastname: Viegelmann
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK. gomoku13@hotmail.com.
authors.forename: Ignacio Juarez
authors.initials: IJ
authors.lastname: Martinez
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK. cheng.cheng1@uni-wuerzburg.de.
authors.forename: Cheng
authors.initials: C
authors.lastname: Cheng
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK. c.dowdells@strath.ac.uk.
authors.forename: Catherine
authors.initials: C
authors.lastname: Dowdells
authors.affiliation: Department of Botany II, Julius-von-Sachs Institute for Biological Sciences, University of Würzburg, Julius-von-Sachs-Platz 3, D-97082 Würzburg, Germany. usama.ramadan@uni-wuerzburg.de.
authors.forename: Usama Ramadam
authors.initials: UR
authors.lastname: Abdelmohsen
authors.affiliation: Department of Botany II, Julius-von-Sachs Institute for Biological Sciences, University of Würzburg, Julius-von-Sachs-Platz 3, D-97082 Würzburg, Germany. christine.gernert@mail.uni-wuerzburg.de.
authors.forename: Christine
authors.initials: C
authors.lastname: Gernert
authors.affiliation: Department of Botany II, Julius-von-Sachs Institute for Biological Sciences, University of Würzburg, Julius-von-Sachs-Platz 3, D-97082 Würzburg, Germany. ute.hentschel@uni-wuerzburg.de.
authors.forename: Ute
authors.initials: U
authors.lastname: Hentschel
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK. ruangelie.edrada-ebel@strath.ac.uk.
authors.forename: RuAngelie
authors.initials: R
authors.lastname: Edrada-Ebel
mesh_terms: D000818:Animals; D001419:Bacteria / Q000302:isolation & purification / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D007448:Invertebrates / Q000382:microbiology*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods; D055432:Metabolomics / Q000379:methods*; D025341:Principal Component Analysis; D012606:Scotland; D064210:Secondary Metabolism; D013559:Symbiosis