Abstract: The present invention relates methods for the biosynthesis of tetrodotoxin (TTX) involving the steps of obtaining a culture possessing one or more of a Vibrio species, such as through a seed culture, inoculating the culture and tissue extract from a textrodotoxin-bearing organism in a fermenter medium, and isolating and purifying tetrodotoxin from said fermenter, resulting in a yield of TTX of about 0.5 g TTX/L in about 3 to 5 days, such TTX being at least 90% pure.