Toxicity and Protein Expression of Alexandrium Species Collected in the Philippine Waters
Bryan John J. Subong1, Garry A. Benico1, Arielle Kae L. Sulit1a,
Christopher O. Mendoza1b, Lourdes J. Cruz1, Rhodora V. Azanza1, and Elsie C. Jimenez2*
1The Marine Science Institute, College of Science,
University of the Philippines Diliman, Quezon City 1101, Philippines
2Department of Physical Sciences, College of Science,
University of the Philippines Baguio, Baguio City 2600, Philippines
Present addresses:
aSt. Luke’s Medical Center, E. Rodriguez Sr. Ave., Quezon City 1112, Philippines
bPhilippine Nuclear Research Institute, Commonwealth Ave., Diliman, Quezon City 1101, Philippines
*Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.
ABSTRACT
Isolates of Alexandrium species collected in the Philippine waters were examined during the exponential growth phase to compare their toxicities and protein expression profiles, and also to correlate protein expression with toxin production. Molecular methods showed that Alexandrium cf. pacificum was genetically divergent from Alexandrium tamarense complex Group IV/ Alexandrium pacificum. Toxin analyses using pre-oxidation method prior to HPLC purification were carried out to determine saxitoxin (STX), neosaxitoxin (neoSTX), and gonyautoxins 1-4 (GTX) levels. This study showed that cultured Alexandrium affine collected in Honda Bay, Palawan produced different STX analogs at various times of culture, which differed from other results showing that this species was non-toxic. The cultured Alexandrium cf. pacificum collected in Anda Channel (ATANDA) was two-fold more toxic than the cultured Alexandrium cf. pacificum collected in Bolinao Channel (ATBOL). Comparative protein expression analyses using 2-D gel electrophoresis were performed for the two Alexandrium cf. pacificum strains (ATANDA and ATBOL) during the exponential growth phase. A unique 2-DE protein spot in ATANDA showed sequence homology with bifunctional ornithine acetyltransferase/ N-acetylglutamate synthase (ArgJ) that has a role in the biosynthesis of arginine, a precursor in STX biosynthesis. The greater detectable expression of such enzyme in the ATANDA strain was correlated with the greater toxicity, suggesting the enzyme’s major participation in toxin biosynthesis.
INTRODUCTION
Research efforts have been geared towards the understanding of the major causative agents of paralytic shellfish poisoning (PSP) in the Philippines due to the numerous cases of PSP leading to human fatalities. It has been reported that majority of PSP cases in Southeast Asia occurred in the Philippines due mainly to blooms of dinoflagellates such as Pyrodinium bahamense and Alexandrium sp. (Yñiguez et al. 2012, Azanza & Benico 2013). . . . . read more
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