title: Production and isolation of azaspiracid-1 and -2 from Azadinium spinosum culture in pilot scale photobioreactors.
abstract: Azaspiracid (AZA) poisoning has been reported following consumption of contaminated shellfish, and is of human health concern. Hence, it is important to have sustainable amounts of the causative toxins available for toxicological studies and for instrument calibration in monitoring programs, without having to rely on natural toxin events. Continuous pilot scale culturing was carried out to evaluate the feasibility of AZA production using Azadinium spinosum cultures. Algae were harvested using tangential flow filtration or continuous centrifugation. AZAs were extracted using solid phase extraction (SPE) procedures, and subsequently purified. When coupling two stirred photobioreactors in series, cell concentrations reached 190,000 and 210,000 cell · mL(-1) at steady state in bioreactors 1 and 2, respectively. The AZA cell quota decreased as the dilution rate increased from 0.15 to 0.3 day(-1), with optimum toxin production at 0.25 day(-1). After optimization, SPE procedures allowed for the recovery of 79 ± 9% of AZAs. The preparative isolation procedure previously developed for shellfish was optimized for algal extracts, such that only four steps were necessary to obtain purified AZA1 and -2. A purification efficiency of more than 70% was achieved, and isolation from 1200 L of culture yielded 9.3 mg of AZA1 and 2.2 mg of AZA2 of >95% purity. This work demonstrated the feasibility of sustainably producing AZA1 and -2 from A. spinosum cultures.
authors: Ifremer, EMP/PHYC Laboratory, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors: Thierry
authors: T
authors: Jauffrais
authors: Marine Institute, Rinville, Oranmore, Co., Galway, Ireland.
authors: Jane
authors: 0000-0002-1219-7731
authors: J
authors: Kilcoyne
authors: Ifremer, EMP/PHYC Laboratory, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors: Véronique
authors: V
authors: Séchet
authors: Nantes Atlantic University, MMS EA2160, 9 rue Bias, 44035 Nantes, France.
authors: Christine
authors: C
authors: Herrenknecht
authors: Ifremer, EMP/PHYC Laboratory, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors: Philippe
authors: P
authors: Truquet
authors: Ifremer, EMP/PHYC Laboratory, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors: Fabienne
authors: 0000-0002-5485-7611
authors: F
authors: Hervé
authors: Ifremer, BRM/PBA Laboratory, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors: Jean Baptiste
authors: JB
authors: Bérard
authors: Marine Institute, Rinville, Oranmore, Co., Galway, Ireland.
authors: Cíara
authors: C
authors: Nulty
authors: Ifremer, EMP/PHYC Laboratory, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors: Sarah
authors: S
authors: Taylor
authors: Alfred Wegener Institute, Am Handelshafen 12, D-27570 Bremerhaven, Germany.
authors: Urban
authors: 0000-0002-8207-4382
authors: U
authors: Tillmann
authors: Norwegian Veterinary Institute, P. O. Box 750 Sentrum, 0106 Oslo, Norway.
authors: Christopher O
authors: CO
authors: Miles
authors: Ifremer, EMP/PHYC Laboratory, Rue de l'Ile d'Yeu, 44311 Nantes, France.
authors: Philipp
authors: 0000-0002-9047-1345
authors: P
authors: Hess
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D004141:Dinoflagellida / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism; D008387:Marine Toxins / Q000096:biosynthesis / Q000302:isolation & purification*; D058433:Photobioreactors; D052616:Solid Phase Extraction / Q000379:methods; D013141:Spiro Compounds / Q000302:isolation & purification*; D014118:Toxins, Biological / Q000096:biosynthesis / Q000302:isolation & purification*; D000095702:Polyether Toxins
chemical_list: D008387:Marine Toxins; D013141:Spiro Compounds; D014118:Toxins, Biological; C406592:azaspiracid; D000095702:Polyether Toxins