Production and Isolation of Azaspiracid-1 and -2 from Azadinium spinosum Culture in Pilot Scale Photobioreactors
|Author(s)||Jauffrais Thierry1, Kilcoyne Jane6, Sechet Veronique1, Herrenknecht Christine2, Truquet Philippe1, Herve Fabienne1, Berard Jean-Baptiste3, Nulty Ciara6, Taylor Sarah1, Tillmann Urban4, Miles Christopher O.5, Hess Philipp1|
|Affiliation(s)||1 : IFREMER, EMP PHYC Lab, F-44311 Nantes, France.
2 : Nantes Atlantic Univ, MMS EA2160, F-44035 Nantes, France.
3 : IFREMER, BRM PBA Lab, F-44311 Nantes, France.
4 : Alfred Wegener Inst, D-27570 Bremerhaven, Germany.
5 : Norwegian Vet Inst, N-0106 Oslo, Norway.
6 : Inst Marine, Oranmore, Co Galway, Ireland.
|Source||Marine Drugs (1660-3397) (Mdpi Ag), 2012-06 , Vol. 10 , N. 6 , P. 1360-1382|
|WOS© Times Cited||23|
|Keyword(s)||solid phase extraction, photobioreactor, chemostat, dinoflagellate, micro-algae, LC-MS/MS, tangential flow filtration, azaspiracid, HP-20|
|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.|