Abstract:
This study explores the expression of Fischerella thermalis Orange Carotenoid Protein 2 (OCP2) as a carrier for ketocarotenoids within the model algae Chlamydomonas reinhardtii. The research addresses the gap in stable expression of carotenoid-binding proteins in green algae to improve ketocarotenoid solubility. The aim was to produce and purify a functional, photoactive FtOCP2 holoprotein, which binds ketocarotenoids like astaxanthin in C. reinhardtii, facilitating their retention in solution. The methodology included codon optimization, transgene expression with chloroplast targeting, protein extraction, and High-Performance Liquid Chromatography (HPLC) analysis. Results showed successful FtOCP2 expression with chloroplast localization and binding to ketocarotenoids, confirmed through spectroscopic analysis. The photoconversion capability of FtOCP2 was observed with ketocarotenoids, enhancing photoprotection. The study demonstrates that C. reinhardtii can serve as a viable production system for soluble ketocarotenoids.
Main Takeaway:
The findings underscore C. reinhardtii as a promising host for producing water-soluble carotenoid-binding proteins that retain antioxidant-rich ketocarotenoids, with potential applications in nutraceuticals and industrial use.
Conclusion on the Role of Constant Systems Cell Disruption equipment:
The Constant Systems Cell Disruptor played a crucial role in this research, facilitating the high-pressure cell lysis required to extract FtOCP2 proteins from algal cells. This efficient disruption process enabled the reliable recovery of target proteins for further purification and analysis, significantly contributing to the successful isolation and functional assessment of FtOCP2 in C. reinhardtii.
