Carotenoid-Rich Algal Biomass for Aquaculture: Astaxanthin Production by Haematococcus Pluvialis

The synthesis of carotenoids has been studied toward enhancing the production of ketocarotenoids, since fish and crustaceans raised by aquaculture require astaxanthin and other ketocaroteinoids in their feed for desirable pigmentation. Notable progress has been made in attaining the goals of determining improved conditions for ketocarotenoid production in Haematococcus pluvialis and in elucidating the carotenoid biosynthetic pathway. For production of astaxanthin a number of strains of the green alga Haematococcus were evaluated, a strain CCAG was found to be optimal for photoautotrophic growth. Of four mutants, selected for enhanced carotenoid production, two hold considerable promise because caroteinoid accumulation occurs without encystment. The biosynthetic pathway of carotenoids was elucidated in photosynthetic organisms by characterizing novel genes encoding carotenoid enzymes and by examining the function of these enzymes in a bacterial complementation system. Two cyclases (b- and e-) were cloned that are at a critical branch point in the pathway. One branch leads to the formation of b-carotene and zeaxanthin and astaxanthin, and the other to the production of a-carotene and lutein. Cyclization of both endgroups of lycopene to yield b-carotene was shown to be catalyzed by a single gene product, b-lycopene cyclase in cyanobacteria and plants. The formation of a-carotene was found to require the e-cyclase gene product in addition to the b-cyclase. By cloning a b-hydroxylase gene we showed that a single gene product forms zeaxanthin by hydroxylatin of both b-carotene rings. It is expected that a second hydroxylase is required in the synthesis of astaxanthin, since canthaxanthin rather than zeaxanthin is the precursor. Evidence, from inhibitor studies, suggests that astaxanthin is formed from canthaxanthin and that b-carotene is a major precursor. Feasibility studies with the photobioreactors have shown that a two-stage system is the most practical, where Haematococcus cultures are first grown to high cell density and are then switched to high light for maximal astaxanthin production. The basic knowledge and molecular tools generated from this study will significantly enhance Haematococcus as a viable model for enhanced astaxanthin production.