Elevated accumulation of lutein and zeaxanthin in a novel high-biomass yielding strain Dunaliella sp. ZP-1 obtained through EMS mutagenesis.

BACKGROUND

Dunaliella microalgae, such as Dunaliella salina riching in β-carotene and Dunaliella bardawil rich in lutein and α-carotene, have been used in aquaculture, supplements, cosmetics, and feed industries. The genus Dunaliella is diverse; therefore, characterization of novel strains and isolation of new varieties through mutagenesis technology will promote natural carotenoid bioproduction.

RESULTS

Salt stress test demonstrated that the newly isolated microalgae strain ZP-1 was a halotolerant strain. Morphology observation and molecular phylogeny analysis indicated that the unicellular green microalga ZP-1 was a member of the genus Dunaliella. Biomass of ZP-1 in RAM medium was up to 2.45 g/L, showing the advantage over other common Dunaliella microalgae in terms of yield. Furthermore, Ethyl methanesulfonate (EMS) mutant library was generated from this high-biomass strain, aiming to improve natural carotenoid productivity. A mutant strain was selected through morphology observation combining with carotenoid quantification by HPLC, which was nominated as turn yellow dunaliella 4 (tyd4). The mutant tyd4 displayed an increased lutein productivity by 28.55% and an increased zeaxanthin productivity by 22.19%. Biomass of tyd4 was promoted by 17.40% through continuous culture under red light. Application of exogenous 1.0 μM melatonin on the mutant tyd4 led to increased cell density and improved biomass.

CONCLUSIONS

Results in this study support that EMS mutagenesis is an effective breeding approach for further improvement of Dunaliella sp. ZP-1, which is a high-biomass microalgae exhibiting potential to overcome the bottleneck of low biomass of current commercial Dunaliella strains. The mutant tyd4 had higher contents of both lutein and zeaxanthin, whose yield could be further elevated by red light and melatonin. This study provided new microalgae sources for scientific research and technical reference for the bioproduction of natural carotenoids.