Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Zeaxanthin Epoxidase 3 Knockout Mutants of the Model Diatom Phaeodactylum tricornutum Enable Commercial Production of the Bioactive Carotenoid Diatoxanthin

Cecilie Græsholt
,
Tore Brembu
ORCID logo ,
Charlotte Volpe
ORCID logo ,
Zdenka Bartosova
ORCID logo ,
Manuel Serif
,
Per Winge
ORCID logo ,
Marianne Nymark
*
Version 1 : Received: 30 March 2024 / Approved: 1 April 2024 / Online: 2 April 2024 (12:46:38 CEST)

A peer-reviewed article of this Preprint also exists.

Græsholt, C.; Brembu, T.; Volpe, C.; Bartosova, Z.; Serif, M.; Winge, P.; Nymark, M. Zeaxanthin epoxidase 3 Knockout Mutants of the Model Diatom Phaeodactylum tricornutum Enable Commercial Production of the Bioactive Carotenoid Diatoxanthin. Mar. Drugs 2024, 22, 185. Græsholt, C.; Brembu, T.; Volpe, C.; Bartosova, Z.; Serif, M.; Winge, P.; Nymark, M. Zeaxanthin epoxidase 3 Knockout Mutants of the Model Diatom Phaeodactylum tricornutum Enable Commercial Production of the Bioactive Carotenoid Diatoxanthin. Mar. Drugs 2024, 22, 185.

Abstract

Antioxidant, anti-inflammatory and chemo-preventive features have been reported for the carotenoid diatoxanthin. Diatoxanthin is only produced by a few groups of microalgae where it functions in photoprotection. Its large-scale production in microalgae is currently not feasible. In fact, rapid conversion into the inactive pigment diadinoxanthin is triggered when the cells are removed from the high-intensity light source, which will be the case during large-scale harvesting of microalgae biomass. Zeaxanthin epoxidase (ZEP) 2 and/or ZEP3 have been suggested to be responsible for the back-conversion of high-light accumulated diatoxanthin to diadinoxanthin in low light in diatoms. Using the CRISPR/Cas9 gene editing technology we knocked out the ZEP2 and ZEP3 genes in the already commercially used marine diatom Phaeodactylum tricornutum to investigate their role in the diadinoxanthin-diatoxanthin cycle, and to determine if one of the mutant strains could function as a diatoxanthin production line. Light shift experiments proved that ZEP3 encodes the enzyme converting diatoxanthin to diadinoxanthin in low light. Loss of ZEP3 caused the high-light accumulated diatoxanthin to be stable for several hours after the cultures had been returned to low light, suggesting that zep3 mutant strains could be suitable as commercial production lines of diatoxanthin.

Keywords

bioactive carotenoid; diatoxanthin; Phaeodactylum tricornutum; CRISPR/Cas9 gene editing; zeaxanthin epoxidase; commercial production line

Subject

Biology and Life Sciences, Biology and Biotechnology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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