27 research outputs found
Dissolved organic matter produced by Thalassiosira pseudonana
Author Posting. Ā© The Author(s), 2014. This is the author's version of the work. It is posted here by permission of Elsevier for personal use, not for redistribution. The definitive version was published in Marine Chemistry 168 (2015): 114-123, doi:10.1016/j.marchem.2014.11.003.Phytoplankton are significant producers of dissolved organic matter (DOM) in marine ecosystems but the identity and dynamics of this DOM remain poorly constrained. Knowledge on the identity and dynamics of DOM are crucial for understanding the molecular-level reactions at the base of the global carbon cycle. Here we apply emerging analytical and computational tools from metabolomics to investigate the composition of DOM produced by the centric diatom Thalassiosira pseudonana. We assessed both intracellular metabolites within T. pseudonana (the endo-metabolome) and extracellular metabolites released by T. pseudonana (the exo-metabolome). The intracellular metabolites had a more variable composition than the extracellular metabolites. We putatively identified novel compounds not previously associated with T. pseudonana as well as compounds that have previously been identified within T. pseudonanaās metabolic capacity (e.g. dimethylsulfoniopropionate and degradation products of chitin). The resulting information will provide the basis for future experiments to assess the impact of T. pseudonana on the composition of dissolved organic matter in marine environments.Instrumentation in the WHOI FT-MS facility was funded by the National Science Foundation MRI program (OCE-0619608) and by the Gordon and Betty T. Moore Foundation (Grant #1214). This work was supported by NSF grant OCE-0928424 to EBK
Organic sulfur: a spatially variable and understudied component of marine organic matter
Ā© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Longnecker, K., Oswald, L., Soule, M. C. K., Cutter, G. A., & Kujawinski, E. B. Organic sulfur: a spatially variable and understudied component of marine organic matter. Limnology and Oceanography Letters, (2020), doi:10.1002/lol2.10149.Sulfur (S) is a major heteroatom in organic matter. This project evaluated spatial variability in the concentration and molecularālevel composition of organic sulfur along gradients of depth and latitude. We measured the concentration of total organic sulfur (TOS) directly from whole seawater. Our data reveal high variability in organic sulfur, relative to established variability in total organic carbon or nitrogen. The deep ocean contained significant amounts of organic sulfur, and the concentration of TOS in North Atlantic Deep Water (NADW) decreased with increasing age while total organic carbon remained stable. Analysis of dissolved organic matter extracts by ultrahigh resolution mass spectrometry revealed that 6% of elemental formulas contained sulfur. The sulfurācontaining compounds were structurally diverse, and showed higher numbers of sulfurācontaining elemental formulas as NADW moved southward. These measurements of organic sulfur in seawater provide the foundation needed to define the factors controlling organic sulfur in the global ocean.We thank Catherine Carmichael, Winifred Johnson, and Gretchen Swarr for assistance with sample collection and processing, and Joe Jennings for the analysis of inorganic nutrients. The help of the captain and crew of the R/V Knorr and the other cruise participants during the āDeepDOMā cruise is appreciated. Two anonymous reviewers and Patricia Soranno provided thorough comments that greatly improved the manuscript. The ultrahigh resolution mass spectrometry samples were analyzed at the WHOI FTāMS Users' Facility that is funded by the National Science Foundation (grant OCEā0619608) and the Gordon and Betty Moore Foundation (GMBF1214). This project was funded by NSF grants OCEā1154320 (to EBK and KL), the W.M. Marquet Award (to KL), and OCEā1435708 (to GAC). The authors declare no conflicts of interest
Extraction efficiency and quantification of dissolved metabolites in targeted marine metabolomics
Ā© The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Limnology and Oceanography: Methods 15 (2017): 417ā428, doi:10.1002/lom3.10181.The field of metabolomics seeks to characterize the suite of small molecules that comprise the end-products of cellular regulation. Metabolomics has been used in biomedical applications as well as environmental studies that explore ecological and biogeochemical questions. We have developed a targeted metabolomics method using electrospray ionizationāliquid chromatography tandem mass spectrometry to analyze metabolites dissolved in seawater. Preparation of samples from the marine environment presents challenges because dilute metabolites must be concentrated and desalted. We present the extraction efficiencies of 89 metabolites in our targeted method using solid phase extraction (SPE). In addition, we calculate the limits of detection and quantification for the metabolites in the method and compare the instrument response factors in five different matrices ranging from deionized water to spent medium from cultured marine microbes. High background organic matter content reduces the instrument response factor for only a small group of metabolites, yet enhances the extraction efficiency for other metabolites on the SPE cartridge used here, a modified styrene-divinylbenzene polymer called PPL. Aromatic or larger uncharged compounds, in particular, are reproducibly well retained on the PPL polymer. This method is suitable for the detection of dissolved metabolites in marine samples, with limits of detection ranging fromā<ā1 pM to ā¼ 2 nM dependent on the dual impacts of seawater matrix on extraction efficiency and on instrument response factors.Gordon and Betty Moore Foundation Grant Number: 3304;
National Science Foundation Grant Number: OCE-1154320;
Simons Foundation Internationa
Evidence for quorum sensing and differential metabolite production by a marine bacterium in response to DMSP
Ā© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in The ISME Journal 10 (2016): 2304ā2316, doi:10.1038/ismej.2016.6.Microbes, the foundation of the marine foodweb, do not function in isolation, but rather rely on molecular level interactions among species to thrive. Although certain types of interactions between autotrophic and heterotrophic microorganisms have been well documented, the role of specific organic molecules in regulating inter-species relationships and supporting growth are only beginning to be understood. Here, we examine one such interaction by characterizing the metabolic response of a heterotrophic marine bacterium, Ruegeria pomeroyi DSS-3, to growth on dimethylsulfoniopropionate (DMSP), an abundant organosulfur metabolite produced by phytoplankton. When cultivated on DMSP, R. pomeroyi synthesized a quorum-sensing molecule, N-(3-oxotetradecanoyl)-l-homoserine lactone, at significantly higher levels than during growth on propionate. Concomitant with the production of a quorum-sensing molecule, we observed differential production of intra- and extracellular metabolites including glutamine, vitamin B2 and biosynthetic intermediates of cyclic amino acids. Our metabolomics data indicate that R. pomeroyi changes regulation of its biochemical pathways in a manner that is adaptive for a cooperative lifestyle in the presence of DMSP, in anticipation of phytoplankton-derived nutrients and higher microbial density. This behavior is likely to occur on sinking marine particles, indicating that this response may impact the fate of organic matter.This research is funded in part by the Gordon and Betty Moore Foundation through Grant GBMF3304 as well as by the National Science Foundation (Grants OCE-0928424 and OCE-1154320)
A phosphate starvation response gene (psr1-like) is present and expressed in Micromonas pusilla and other marine algae
Ā© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Fiore, C. L., Alexander, H., Soule, M. C. K., & Kujawinski, E. B. A phosphate starvation response gene (psr1-like) is present and expressed in Micromonas pusilla and other marine algae. Aquatic Microbial Ecology, 86, (2021): 29ā46, https://doi.org/10.3354/ame01955.Phosphorus (P) limits primary production in regions of the surface ocean, and many plankton species exhibit specific physiological responses to P deficiency. The metabolic response of Micromonas pusilla, an ecologically relevant marine photoautotroph, to P deficiency was investigated using metabolomics and comparative genomics. The concentrations of some intracellular metabolites were elevated in the P-deficient cells (e.g. xanthine, inosine), and genes involved in the associated metabolic pathways shared a predicted conserved amino acid motif in the non-coding regions of each gene. The presence of the conserved motif suggests that these genes may be co-regulated, and the motif may constitute a regulatory element for binding a transcription factor, specifically that of Psr1 (phosphate starvation response). A putative phosphate starvation response gene ( psr1-like) was identified in M. pusilla with homology to well characterized psr1/ phr1 genes in algae and plants, respectively. This gene appears to be present and expressed in other marine algal taxa (e.g. Emiliania huxleyi) in field sites that are chronically P limited. Results from the present study have implications for understanding phytoplankton taxon-specific roles in mediating P cycling in the ocean.This research was funded by the Gordon and Betty Moore Foundation through Grant GBMF3304 to E.B.K., and it was partially supported by a grant from the Simons Foundation (Award ID 509034 to E.B.K.)
Release of ecologically relevant metabolites by the cyanobacterium Synechococcus elongatusā CCMP 1631
Author Posting. Ā© The Author(s), 2015. This is the author's version of the work. It is posted here by permission of Society for Applied Microbiology for personal use, not for redistribution. The definitive version was published in Environmental Microbiology 17 (2015): 3949ā3963, doi:10.1111/1462-2920.12899.Photoautotrophic plankton in the surface ocean release organic compounds that fuel secondary production by heterotrophic bacteria. Here we show that an abundant marine cyanobacterium, Synechococcus elongatus, contributes a variety of nitrogen-rich and sulfur-containing compounds to dissolved organic matter. A combination of targeted and untargeted metabolomics and genomic tools was used to characterize the intracellular and extracellular metabolites of S. elongatus. Aromatic compounds such as 4-hydroxybenzoic acid and phenylalanine, as well as nucleosides (e.g., thymidine, 5ā-methylthioadenosine, xanthosine), the organosulfur compound 3-mercaptopropionate, and the plant auxin indole 3-acetic acid, were released by S. elongatus at multiple time points during its growth. Further, the amino acid kynurenine was found to accumulate in the media even though it was not present in the predicted metabolome of S. elongatus. This indicates that some metabolites, including those not predicted by an organismās genome, are likely excreted into the environment as waste; however, these molecules may have broader ecological relevance if they are labile to nearby microbes. The compounds described herein provide excellent targets for quantitative analysis in field settings to assess the source and lability of dissolved organic matter in situ.This project was funded by the Gordon and Betty Moore Foundation through Grant #3304 to E. Kujawinski.2016-07-0
Hepatic metabolite profiling of polychlorinated biphenyl (PCB)-resistant and sensitive populations of Atlantic killifish (Fundulus heteroclitus)
Author Posting. Ā© The Author(s), 2018. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Aquatic Toxicology (2018), doi:10.1016/j.aquatox.2018.10.007.Atlantic killifish inhabiting polluted sites along the east coast of the U.S. have evolved
resistance to toxic effects of contaminants. One such contaminated site is the Acushnet River
estuary, near New Bedford Harbor (NBH), Massachusetts, which is characterized by very high
PCB concentrations in the sediments and in the tissues of resident killifish. Though killifish at
this site appear to be thriving, the metabolic costs of survival in a highly contaminated
environment are not well understood. In this study we compared the hepatic metabolite profiles
of resistant (NBH) and sensitive populations (Scorton Creek (SC), Sandwich, MA) using a
targeted metabolomics approach in which polar metabolites were extracted from adult fish livers
and quantified. Our results revealed differences in the levels of several metabolites between fish
from the two sites. The majority of these metabolites are associated with one-carbon
metabolism, an important pathway that supports multiple physiological processes including DNA
and protein methylation, nucleic acid biosynthesis and amino acid metabolism. We measured
the gene expression of DNA methylation (DNA methyltransferase 1, dnmt1) and demethylation
genes (Ten-Eleven Translocation (TET) genes) in the two populations, and observed lower
levels of dnmt1 and higher levels of TET gene expression in the NBH livers, suggesting possible
differences in DNA methylation profiles. Consistent with this, the two populations differed
significantly in the levels of 5-methylcytosine and 5-hydroxymethylcytosine nucleotides. Overall,
our results suggest that the unique hepatic metabolite signatures observed in NBH and SC
reflect the adaptive mechanisms for survival in their respective habitats.This work was supported by the Joint Initiative Awards Fund from the Andrew W. Mellon
Foundation (NA and EBK) and National Institute of Environmental Health Sciences (NIEHS)
Superfund Research Program (P42ES007381) at Boston University. LG was supported by the
Postdoctoral Scholar Program at the Woods Hole Oceanographic Institution (with funding
provided by the Townsend Postdoctoral Scholarship Fund, and the John H. Steele Endowment
in support of Postdoctoral Research)
Environmental metabolomics : databases and tools for data analysis
Ā© The Author(s), 2015. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Marine Chemistry 177 (2015): 366ā373, doi:10.1016/j.marchem.2015.06.012.Metabolomics is the study of small molecules, or āmetabolitesā, that are the end products of biological processes. While -omics technologies such as genomics, transcriptomics, and proteomics measure the metabolic potential of organisms, metabolomics provides detailed information on the organic compounds produced during metabolism and found within cells and in the environment. Improvements in analytical techniques have expanded our understanding of metabolomics and developments in computational tools have made metabolomics data accessible to a broad segment of the scientific community. Yet, metabolomics methods have only been applied to a limited number of projects in the marine environment. Here, we review analysis techniques for mass spectrometry data and summarize the current state of metabolomics databases. We then describe a boutique database developed in our laboratory for efficient data analysis and selection of mass spectral targets for metabolite identification. The code to implement the database is freely available on GitHub (https://github.com/joefutrelle/domdb). Data organization and analysis are critical, but often under-appreciated, components of metabolomics research. Future advances in environmental metabolomics will take advantage of continued development of new tools that facilitate analysis of large metabolomics datasets.The field data populating the database comes from scientific cruises funded by grants from the National Science Foundation to EBK and KL (Atlantic Ocean, OCE-1154320) and E.V. Armbrust (Pacific Ocean, OCE-1205233). The laboratory experiment with coastal seawater was funded by a grant from the Gulf of Mexico Research Initiative to EBK and H.K. White. The laboratory experiments with microbial isolates and the database development are funded by the Gordon and Betty Moore Foundation through Grant GBMF3304 to EBK
Organic Sulfur: A Spatially Variable and Understudied Component of Marine Organic Matter
Sulfur (S) is a major heteroatom in organic matter. This project evaluated spatial variability in the concentration and molecular-level composition of organic sulfur along gradients of depth and latitude. We measured the concentration of total organic sulfur (TOS) directly from whole seawater. Our data reveal high variability in organic sulfur, relative to established variability in total organic carbon or nitrogen. The deep ocean contained significant amounts of organic sulfur, and the concentration of TOS in North Atlantic Deep Water (NADW) decreased with increasing age while total organic carbon remained stable. Analysis of dissolved organic matter extracts by ultrahigh resolution mass spectrometry revealed that 6% of elemental formulas contained sulfur. The sulfurcontaining compounds were structurally diverse, and showed higher numbers of sulfur-containing elemental formulas as NADW moved southward. These measurements of organic sulfur in seawater provide the foundation needed to define the factors controlling organic sulfur in the global ocean
Extracellular reef metabolites across the protected Jardines de la Reina, Cuba Reef System
Ā© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Weber, L., Armenteros, M., Soule, M. K., Longnecker, K., Kujawinski, E. B., & Apprill, A. Extracellular reef metabolites across the protected Jardines de la Reina, Cuba Reef System. Frontiers in Marine Science, 7, (2020): 582161, https://doi.org/10.3389/fmars.2020.582161.Coral reef ecosystems are incredibly diverse marine biomes that rely on nutrient cycling by microorganisms to sustain high productivity in generally oligotrophic regions of the ocean. Understanding the composition of extracellular reef metabolites in seawater, the small organic molecules that serve as the currency for microorganisms, may provide insight into benthic-pelagic coupling as well as the complexity of nutrient cycling in coral reef ecosystems. Jardines de la Reina (JR), Cuba is an ideal environment to examine extracellular metabolites across protected and high-quality reefs. Here, we used liquid chromatography mass spectrometry (LC-MS) to quantify specific known metabolites of interest (targeted metabolomics approach) and to survey trends in metabolite feature composition (untargeted metabolomics approach) from surface and reef depth (6 ā 14 m) seawater overlying nine forereef sites in JR. We found that untargeted metabolite feature composition was surprisingly similar between reef depth and surface seawater, corresponding with other biogeochemical and physicochemical measurements and suggesting that environmental conditions were largely homogenous across forereefs within JR. Additionally, we quantified 32 of 53 detected metabolites using the targeted approach, including amino acids, nucleosides, vitamins, and other metabolic intermediates. Two of the quantified metabolites, riboflavin and xanthosine, displayed interesting trends by depth. Riboflavin concentrations were higher in reef depth compared to surface seawater, suggesting that riboflavin may be produced by reef organisms at depth and degraded in the surface through photochemical oxidation. Xanthosine concentrations were significantly higher in surface reef seawater. 5ā²-methylthioadenosine (MTA) concentrations increased significantly within the central region of the archipelago, displaying biogeographic patterns that warrant further investigation. Here we lay the groundwork for future investigations of variations in metabolite composition across reefs, sources and sinks of reef metabolites, and changes in metabolites over environmental, temporal, and reef health gradients.This work was supported by the Dalio Foundation (now āOceanXā) and the National Science Foundation (OCE-1736288) (award to Amy Apprill). The mass spectrometry samples were analyzed at the WHOI FT-MS Usersā Facility with instrumentation funded by the National Science Foundation (grant OCE-1058448 to EK and MK) and the Simons Foundation (Award ID #509042, EK). Lastly, a portion of the publication fees was supported by the Massachusetts Institute of Technology (MIT) Open Access Article Publication Subvention fund from MIT Libraries