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  • 1
    Publication Date: 2022-05-26
    Description: Author Posting. © The Author(s), 2013. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Marine Chemistry 154 (2013): 34–45, doi:10.1016/j.marchem.2013.04.003.
    Description: Dissolved organic matter (DOM) is the largest active organic carbon reservoir in the ocean (662 Gt C), a major fraction (〉 95%) of which remains chemically uncharacterized. The concentration and isolation of DOM from seawater by ultrafiltration facilitates its chemical characterization by spectroscopic techniques. Using ultrafiltration, silver cation preparative chromatography and gas chromatography coupled with mass spectrometry (GC-MS), we identified 50 novel sugar compounds after hydrolysis of the high molecular weight dissolved organic matter fraction (HMWDOM; the fraction of DOM isolated after ultrafiltration). Sugars were identified by comparison of their mass spectra with those of chemically synthetized standards and with spectra previously described in the literature. Our results showed that mono- and di- methylated hexoses; mono- and di- methylated pentoses; mono- and di- methylated 6-deoxysugars, as well as heptoses, methylated heptoses, 3,6-dideoxysugars and 1,6 anhydrosugars (levoglucosan, mannosan, and galactosan) are components of HMWDOM, which may explain the low apparent yields of sugars recovered by molecular level (HPLC) analyses of HMWDOM after hydrolysis. From three depths spanning the surface (15 m) to bathypelagic (1800 m) ocean in the North Pacific near Hawaii our results showed that mono- and di- methylated hexoses were most abundant in the surface sample (64% of the total identified methylated sugarcompounds), while at 1800m monomethylated 6-deoxy sugars were the dominant sugars (42% of the total identified methylated sugar compounds). The high diversity of mono- and di- methylated hexoses in the surface sample most likely suggests an algal and/or bacterial source, while the high abundance of methylated 6-deoxy hexoses in the deep sample points toward an important bacterial contribution because the latter sugars are mostly found in bacterial lipopolysaccharides as well as highly degraded organic material.
    Description: Funding was provided by CNRS/INSU LEFE/Cyber grant (DORADE project), region Provence Alpes Côte d’Azur (MANDARINE project) and the Aix Marseille University (ORANGE project). D.J.R. received support from the National Science Foundation Center for Microbial Research and Education (CMORE) DBI 0424599 and the Gordon and Betty Moore Foundation.
    Keywords: High molecular weight dissolved organic matter (HMWDOM) ; Acyl polysaccharides (APS) ; Methylated sugars ; Heptoses ; Anhydrosugars ; 3,6 dideoxysugars ; Chemical synthesis of methylated sugars ; GC-MS
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
    Format: application/pdf
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  • 2
    Publication Date: 2022-10-26
    Description: © The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Li, J., Boiteau, R. M., Babcock-Adams, L., Acker, M., Song, Z., McIlvin, M. R., & Repeta, D. J. Element-selective targeting of nutrient metabolites in environmental samples by inductively coupled plasma mass spectrometry and electrospray ionization mass spectrometry. Frontiers in Marine Science, 8, (2021): 630494, https://doi.org/10.3389/fmars.2021.630494.
    Description: Metabolites that incorporate elements other than carbon, nitrogen, hydrogen and oxygen can be selectively detected by inductively coupled mass spectrometry (ICPMS). When used in parallel with chromatographic separations and conventional electrospray ionization mass spectrometry (ESIMS), ICPMS allows the analyst to quickly find, characterize and identify target metabolites that carry nutrient elements (P, S, trace metals; “nutrient metabolites”), which are of particular interest to investigations of microbial biogeochemical cycles. This approach has been applied to the study of siderophores and other trace metal organic ligands in the ocean. The original method used mass search algorithms that relied on the ratio of stable isotopologues of iron, copper and nickel to assign mass spectra collected by ESIMS to metabolites carrying these elements detected by ICPMS. However, while isotopologue-based mass assignment algorithms were highly successful in characterizing metabolites that incorporate some trace metals, they do not realize the whole potential of the ICPMS/ESIMS approach as they cannot be used to assign the molecular ions of metabolites with monoisotopic elements or elements for which the ratio of stable isotopes is not known. Here we report a revised ICPMS/ESIMS method that incorporates a number of changes to the configuration of instrument hardware that improves sensitivity of the method by a factor of 4–5, and allows for more accurate quantitation of metabolites. We also describe a new suite of mass search algorithms that can find and characterize metabolites that carry monoisotopic elements. We used the new method to identify siderophores in a laboratory culture of Vibrio cyclitrophicus and a seawater sample collected in the North Pacific Ocean, and to assign molecular ions to monoisotopic cobalt and iodine nutrient metabolites in extracts of a laboratory culture of the marine cyanobacterium Prochorococcus MIT9215.
    Description: This work was generously supported by the National Science Foundation grant OCE-1829761 to RB and OCE-1356747 and -1736280 to DR. DR also received generous support from the Simons Foundation Life Sciences Project Award 49476.
    Keywords: LC-MS ; Algorithm ; Environmental metabolomics ; Trace metal ; Siderophores
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 3
    Publication Date: 2022-10-26
    Description: © The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Kiang, N. Y., Swingley, W. D., Gautam, D., Broddrick, J. T., Repeta, D. J., Stolz, J. F., Blankenship, R. E., Wolf, B. M., Detweiler, A. M., Miller, K. A., Schladweiler, J. J., Lindeman, R., & Parenteau, M. N. Discovery of chlorophyll d: isolation and characterization of a far-red cyanobacterium from the original site of manning and strain (1943) at Moss Beach, California. Microorganisms, 10(4), (2022): 819, https://doi.org/10.3390/microorganisms10040819.
    Description: We have isolated a chlorophyll-d-containing cyanobacterium from the intertidal field site at Moss Beach, on the coast of Central California, USA, where Manning and Strain (1943) originally discovered this far-red chlorophyll. Here, we present the cyanobacterium’s environmental description, culturing procedure, pigment composition, ultrastructure, and full genome sequence. Among cultures of far-red cyanobacteria obtained from red algae from the same site, this strain was an epiphyte on a brown macroalgae. Its Qyin vivo absorbance peak is centered at 704–705 nm, the shortest wavelength observed thus far among the various known Acaryochloris strains. Its Chl a/Chl d ratio was 0.01, with Chl d accounting for 99% of the total Chl d and Chl a mass. TEM imagery indicates the absence of phycobilisomes, corroborated by both pigment spectra and genome analysis. The Moss Beach strain codes for only a single set of genes for producing allophycocyanin. Genomic sequencing yielded a 7.25 Mbp circular chromosome and 10 circular plasmids ranging from 16 kbp to 394 kbp. We have determined that this strain shares high similarity with strain S15, an epiphyte of red algae, while its distinct gene complement and ecological niche suggest that this strain could be the closest known relative to the original Chl d source of Manning and Strain (1943). The Moss Beach strain is designated Acaryochloris sp. (marina) strain Moss Beach.
    Description: N.Y.K., M.N.P. and R.E.B. were supported by the NASA Virtual Planetary Laboratory team (VPL), which was funded under NASA Astrobiology Institute Cooperative Agreement Number NNA13AA93A, and Grant Number 80NSSC18K0829. This work also benefited from participation in the NASA Nexus for Exoplanet Systems Science (NExSS) research coordination network (RCN). W.D.S, N.Y.K. and M.N.P. were also supported by a NASA Exobiology grant No. 80NSSC19K0478. J.TB. was supported by the NASA Postdoctoral Program (NPP) award number NPP168014S. N.Y.K. received training support from the NASA Goddard Space Flight Center Training Office to take the Microbial Diversity course at the Marine Biological Laboratory, Woods Hole, MA, USA.
    Keywords: Chlorophyll d ; Acaryochloris ; Moss Beach ; Cyanobacteria ; Far-red photosynthesis ; Photosynthetic pigments ; Absorbance spectra ; Genome sequence
    Repository Name: Woods Hole Open Access Server
    Type: Article
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