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Exploring the high light stress response of a Haematococcus pluvialis (Chlorophyceae) mutant with a higher astaxanthin content via proteomic and phosphoproteomic analysis reveals altered chlorophyll biosynthesis, carbon partitioning, and astaxanthin biosynthesis and trafficking

ABSTRACT: While the green microalga, Haematococcus pluvialis, is the best natural producer of astaxanthin, efforts to grow it for astaxanthin production are still limited by its low biomass productivity which subsequently results in lower astaxanthin productivity. Despite new methods of direct strain modification to improve growth and astaxanthin production, the limited understanding of how the astaxanthin biosynthesis pathway is regulated has been a significant roadblock to strain improvement. This work aims to address this knowledge gap by exploring the stress response of H. pluvialis during the transition from heterotrophy to high light conditions. A physical radiation (heavy-ion beam) mutagenesis approach was applied to generate H. pluvialis mutants with a higher cell division rate under heterotrophic conditions. The mutant JWHIB 27-38 that grew 25% faster than the wild type was identified. Mutant JWHIB 27-38 also achieved a 69.61% higher lipid content per cell and 86.17% higher astaxanthin per cell than the wild type under high light stress conditions. A proteomics and phosphoproteomics analysis was conducted with JWHIB 27-38 and the wild type at 0, 48, and 72 h of high light stress to elucidate the mechanisms underlying these phenotypes. Maintained upregulation of chlorophyll biosynthesis pathway proteins and phosphoproteins in mutant JWHIB 27-38 during high light stress, such as glutamate-1-semialdehyde 2,1-aminomutase (GSAM) and coproporphyrinogen oxidase (CPOX), may support continued chlorophyll biosynthesis which can allow cells to adapt to high light conditions. Upregulation of fatty acid biosynthetic proteins such as biotin carboxyl carrier protein (BCCP) and 3-ketoacyl-CoA synthase (KCS) in mutant JWHIB 27-38 suggests that carbon is favored toward lipid accumulation. Increased lipid content, along with upregulation of the key astaxanthin biosynthesis protein, phytoene synthase (PSY), may enable the higher astaxanthin content in mutant JWHIB 27-38. Additionally, upregulation of a putative astaxanthin-trafficking protein, AstaP, likely enables the mutant to traffic more astaxanthin to the cell periphery as part of its stress response. These and other significantly differentially expressed proteins may provide promising targets for future strain engineering work to improve astaxanthin productivity.

INSTRUMENT(S):

ORGANISM(S): Haematococcus Lacustris

SUBMITTER: Kyarii Ramarui

LAB HEAD: Yantao Li

PROVIDER: PXD060374 | Pride | 2026-01-24

REPOSITORIES: Pride

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			Action	DRS
	PhosphoproteomicsAndromeda.zip	Other
	PhosphoproteomicsSearch.zip	Other
	PhosphoproteomicsmzTab.mzTab	Mztab
	ProteomicsAndromeda.zip	Other
	ProteomicsSearch.zip	Other

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Exploring the high light stress response of a Haematococcus pluvialis (Chlorophyceae) mutant with a higher astaxanthin content via proteomic and phosphoproteomic analysis reveals altered chlorophyll biosynthesis, carbon partitioning, and astaxanthin biosynthesis and trafficking

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