Chemodiversity of Ladder-Frame Prymnesin Polyethers in Prymnesium parvum.

Blooms of the microalga Prymnesium parvum cause devastating fish kills worldwide, which are suspected to be caused by the supersized ladder-frame polyether toxins prymnesin-1 and -2. These toxins have, however, only been detected from P. parvum in rare cases since they were originally described two decades ago. Here, we report the isolation and characterization of a novel B-type prymnesin, based on extensive analysis of 2D- and 3D-NMR data of natural as well as 90% (13)C enriched material. B-type prymnesins lack a complete 1,6-dioxadecalin core unit, which is replaced by a short acyclic C2 linkage compared to the structure of the original prymnesins. Comparison of the bioactivity of prymnesin-2 with prymnesin-B1 in an RTgill-W1 cell line assay identified both compounds as toxic in the low nanomolar range. Chemical investigations by liquid chromatography high-resolution mass spectrometry (LC-HRMS) of 10 strains of P. parvum collected worldwide showed that only one strain produced the original prymnesin-1 and -2, whereas four strains produced the novel B-type prymnesin. In total 13 further prymnesin analogues differing in their core backbone and chlorination and glycosylation patterns could be tentatively detected by LC-MS/HRMS, including a likely C-type prymnesin in five strains. Altogether, our work indicates that evolution of prymnesins has yielded a diverse family of fish-killing toxins that occurs around the globe and has significant ecological and economic impact.

[1]  K. Nielsen,et al.  Chemical Diversity, Origin, and Analysis of Phycotoxins. , 2016, Journal of natural products.

[2]  D. Roelke,et al.  A chronicle of a killer alga in the west: ecology, assessment, and management of Prymnesium parvum blooms , 2015, Hydrobiologia.

[3]  K. Nielsen,et al.  Prymnesium parvum revisited: relationship between allelopathy, ichthyotoxicity, and chemical profiles in 5 strains. , 2014, Aquatic toxicology.

[4]  M. Satake,et al.  Polyketide biosynthesis in dinoflagellates: what makes it different? , 2014, Natural product reports.

[5]  S. Manning,et al.  Isolation of polyketides from Prymnesium parvum (Haptophyta) and their detection by liquid chromatography/mass spectrometry metabolic fingerprint analysis. , 2013, Analytical biochemistry.

[6]  Vivian R. Dayeh,et al.  The Use of Fish‐Derived Cell Lines for Investigation of Environmental Contaminants: An Update Following OECD's Fish Toxicity Testing Framework No. 171 , 2013, Current protocols in toxicology.

[7]  P. Moeller,et al.  Identification of toxic fatty acid amides isolated from the harmful alga Prymnesium parvum carter , 2012 .

[8]  P. Moeller,et al.  The contribution of fatty acid amides to Prymnesium parvum Carter toxicity , 2012 .

[9]  M. Satake,et al.  Brevisulcenal-F: a polycyclic ether toxin associated with massive fish-kills in New Zealand. , 2012, Journal of the American Chemical Society.

[10]  M. Satake,et al.  Complete 13C-labeling pattern of yessotoxin a marine ladder-frame polyether , 2011 .

[11]  R. Cichewicz,et al.  Reassessing the ichthyotoxin profile of cultured Prymnesium parvum (golden algae) and comparing it to samples collected from recent freshwater bloom and fish kill events in North America. , 2010, Toxicon : official journal of the International Society on Toxinology.

[12]  C. Tomas,et al.  Structure and biosynthesis of amphidinol 17, a hemolytic compound from Amphidinium carterae. , 2010, Journal of natural products.

[13]  Gregory M. Southard,et al.  Prymnesium parvum: The Texas Experience 1 , 2010 .

[14]  Loraine T. Fries,et al.  Aspects of The Origins, Ecology, And Control Of Golden Alga Prymnesium parvum: Introduction To The Featured Collection 1 , 2010 .

[15]  W. Eikrem,et al.  Prymnesium parvum: The Norwegian Experience 1 , 2010 .

[16]  O. Fiehn,et al.  FiehnLib: mass spectral and retention index libraries for metabolomics based on quadrupole and time-of-flight gas chromatography/mass spectrometry. , 2009, Analytical chemistry.

[17]  Ryuichi Watanabe,et al.  Synthesis of the JK/LM-ring model of prymnesins, potent hemolytic and ichthyotoxic polycyclic ethers isolated from the red tide alga Prymnesium parvum: confirmation of the relative configuration of the K/L-ring juncture , 2006 .

[18]  M. Satake,et al.  Synthesis of the CDE/FG ring models of prymnesins: reassignment of the relative configuration of the E/F ring juncture. , 2004, Organic letters.

[19]  Takeshi Shida,et al.  Synthesis and stereochemical confirmation of the HI/JK ring system of prymnesins, potent hemolytic and ichthyotoxic glycoside toxins isolated from the red tide alga , 2001 .

[20]  M. Satake,et al.  Absolute configuration at C14 and C85 in prymnesin-2, a potent hemolytic and ichthyotoxic glycoside isolated from the red tide alga Prymnesium parvum. , 2001, Chirality.

[21]  T. Yasumoto,et al.  The structure elucidation and biological activities ofhigh molecular weight algal toxins: maitotoxin, prymnesins andzooxanthellatoxins , 2000 .

[22]  M. Satake,et al.  Structures and Partial Stereochemical Assignments for Prymnesin-1 and Prymnesin-2: Potent Hemolytic and Ichthyotoxic Glycosides Isolated from the Red Tide Alga Prymnesium parvum , 1999 .

[23]  M. Satake,et al.  Prymnesin-2: A Potent Ichthyotoxic and Hemolytic Glycoside Isolated from the Red Tide Alga Prymnesium parvum , 1996 .

[24]  Christian Pedersen,et al.  Carbon-13 Nuclear Magnetic Resonance Spectroscopy of Monosaccharides , 1983 .

[25]  W. König,et al.  Gas Chromatographic Separation of Carbohydrate Enantiomers on a New Chiral Stationary Phase , 1981 .

[26]  Iver W. Duedall,et al.  PREPARATION OF ARTIFICIAL SEAWATER1 , 1967 .

[27]  M. Shilo,et al.  A SENSITIVE ASSAY SYSTEM FOR DETERMINATION OF THE ICHTHYOTOXICITY OF PRYMNESIUM PARVUM. , 1964, Journal of general microbiology.