Title: NMR-Based Microbial Metabolomics and the temperature-dependent coral pathogen Vibrio coralliilyticus

Arezue F. B. Boroujerdi,^1,2 Alexander Meyers,³ Elizabeth C. Pollock,⁴ Sara Lien Huynh,⁵ Tracey Schock,^1,2 Maria Vizcaino^2,6, Pamela J. Morris^2,6, Daniel W. Bearden^1,2

¹National Institute of Standards and Technology, Hollings Marine Laboratory, Charleston, SC 29412

²Hollings Marine Laboratory, Charleston, SC 29412

³Tennessee Technological University, Undergraduate Program in Chemical Engineering. Cookeville, TN 38505

⁴Chemistry Program, The Richard Stockton College of New Jersey, Pomona, NJ 08240

⁵Mt. Holyoke College, Undergraduate Program in Biology. South Hadley, MA 01075

⁶Marine Biomedicine and Environmental Sciences, Medical University of South Carolina, Charleston, SC 29412

Coral bleaching occurs when the symbioses between coral animals and their zooxanthellae is disrupted, either as part of a natural cycle or as the result of unusual events. The bacteria Vibrio coralliilyticus has been shown to cause bleaching in the coral Pocillopora damicornis at temperatures higher than 27 °C.¹ This temperature-dependence of V. coralliilyticus in regard to its metabolome was addressed. One-dimensional nuclear magnetic resonance (NMR) proton spectra were obtained of methanol-water extracts of intracellular metabolites (the endo-metabolome) from multiple samples of the bacteria cultured into late stationary phase at both a high temperature (27 °C) and a low temperature (24 °C). The spectra were subjected to principal components analysis (PCA), and significant temperature-based separations in PC1, PC2, and PC3 dimensions were observed. Betaine, succinate, and glutamate were identified as metabolites in the V. coralliilyticus samples that cause the greatest temperature-based separations in the PC scores plots. With increasing temperature, betaine was shown to be down regulated and glutamate up regulated. These metabolites function as compatible solutes by helping the cells cope with salt from their seawater environment.

[1] Ben-Haim, Y., M. Zicherman-Keren, and E. Rosenberg. 2003. Temperature-regulated bleaching and lysis of the coral Pocillopora damicornis by the novel pathogen Vibrio coralliilyticus. Appl. and Environ. Microbiol. 69(7):4236-42.

CATEGORY: Chemistry

Mentors Name: Daniel W. Bearden

Division 839 (Analytical Chemistry) Group 00 (NMR), CSTL, Hollings Marine Laboratory, 331 Ft. Johnson Rd., Charleston, SC 29412

Tel: (843)762-8924

Fax: (843)762-8742

Email: arezue.boroujerdi@nist.gov

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