Convenient large-scale purification of yessotoxin from Protoceratium reticulatum culture and isolation of a novel furanoyessotoxin.
Yessotoxins from a large-scale culture (226 L) of Protoceratium reticulatum strain CAWD129 were harvested by filtration followed by solid-phase extraction. The extract was purified by column chromatography over basic alumina and reverse-phase flash chromatography to afford pure yessotoxin (193 mg). Isolation of yessotoxin was greatly facilitated by selection of a strain which did not produce analogues that interfered with yessotoxin isolation. In addition to yessotoxin, numerous minor yessotoxins were detected by LC-MS in other fractions. From one of these, an early eluting minor analogue with the same molecular weight as yessotoxin and a similar mass spectrometric fragmentation pattern was isolated. This analogue was identified by NMR and mass spectrometry as a novel yessotoxin analogue containing a furan ring in the side chain. This finding reveals biosynthetic flexibility of the yessotoxin pathway in P. reticulatum and confirms earlier findings of production of many minor yessotoxin analogues by this alga. Production of these analogues appeared to be a constitutive trait of P. reticulatum CAWD129.
AgResearch Ltd., Ruakura Research Centre, Hamilton, New Zealand.
Jared I
JI
Loader
Allan D
AD
Hawkes
Veronica
V
Beuzenberg
Dwayne J
DJ
Jensen
Janine M
JM
Cooney
Alistair L
AL
Wilkins
Joan M
JM
Fitzgerald
Lyn R
LR
Briggs
Christopher O
CO
Miles
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004141:Dinoflagellida / Q000378:metabolism*; D004988:Ethers, Cyclic / Q000737:chemistry / Q000302:isolation & purification*; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D008978:Mollusk Venoms; D035964:Oxocins / Q000737:chemistry / Q000302:isolation & purification*; D021241:Spectrometry, Mass, Electrospray Ionization
D004988:Ethers, Cyclic; D008978:Mollusk Venoms; D035964:Oxocins; C526859:furanoyessotoxin; C066632:yessotoxin