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| Main Authors: | , , , , |
|---|---|
| Format: | Artículo científico |
| Language: | en |
| Published: |
Marine drugs
2025
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| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/41440904/ |
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Table of Contents:
- Installing a Ketocarotenoid Branch in via Functional Activation of β-Carotene Ketolase. Chao, Hengshen Kamal, Rasool Wu, Yan Huang, Danqiong Wang, Chaogang Chlamydomonas reinhardtii Diatoms Xanthophylls Metabolic Engineering Oxygenases Carotenoids Astaxanthin is a high-value ketocarotenoid antioxidant, but its industrial production from is constrained by multi-stage cultivation and a rigid cell wall that hinders downstream extraction. The marine diatom , which lacks these limitations, represents a promising alternative chassis because it grows fast, lacks a recalcitrant wall, and supports efficient pigment accumulation. This study establishes a functional ketocarotenoid biosynthetic branch in through rational metabolic engineering. To address challenges in protein targeting posed by the host's complex plastid architecture, we performed heterologous expression of the β-carotene ketolase (CrBKT), fused at its N-terminus to bipartite transit peptides derived from two endogenous proteins. Western blotting and UPLC-MS/MS analysis confirmed that only the transit peptide fused constructs produced stable protein and functional activity, whereas the native CrBKT failed. The rationally engineered strain successfully accumulated ~45 µg/g DCW of canthaxanthin and ~15 µg/g DCW of astaxanthin. Metabolomic profiling revealed a 50% reduction in fucoxanthin, indicating a substantial redirection of metabolic flux from the native pathway toward the engineered ketocarotenoid branch. This work establishes as a viable platform for ketocarotenoid production and highlights the critical role of evolution-aware plastid targeting in heterologous pathway reconstruction within complex algal systems.