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<p>As a remarkable model protein for studying excitation energy transfer (EET), Cryptophyte phycocyanin 645 (Cr-PC645) is not only important in the frontier research field on photosynthesis but also, like other kinds of phycocyanin, valuable in food and cosmetic industries as a kind of natural blue pigment protein. Here, we report a convenient preparation method for the purification of Cr-PC645 from cryptophytic algae, <i>Chroomonas placoidea</i>, mainly using hydrophobic interaction chromatography, and a study of EET characterization based on it. The final recovery of Cr-PC645 is 58.49% and the highest purity of final samples reaches 7.27 (<i>A</i><sub>645</sub>/<i>A</i><sub>280</sub>) in the obtained products. In addition to high purity, the properties of unchanged intact optical activity of purified Cr-PC645 are proved by comprehensively analyzing absorption, fluorescence, circular dichroism, and ultrafast time-resolved spectra. Furthermore, three time components of 6, 59, and 1021 ps are confirmed by 77&#xa0;K time-resolved fluorescence spectroscopy experiments, which can be assigned to the time of energy transfer process between specific phycocyanobilins (PCBs), suggesting that the excitation energy is transferred from PCB-158 to PCB-82 in the same β subunit and localized the PCB-82 chromophores between different β subunits, then transferred to the downstream acceptors. In view of all these results, this optimized method is efficient and will be conducive to multi-field applications in the future.</p> Graphical Abstract <p></p>

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Convenient Preparation and Energy Transfer Kinetic Process Verification by Ultrafast Time-resolved Fluorescence Spectra for Natural Cryptophyte Phycocyanin 645

  • 期刊论文

  • 发表日期：

    2026年2月9日

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  • Yang Pu,
  • Chuanxu Liu,
  • Xiangyu Lan,
  • Chao Xiao,
  • Mingyuan Xie,
  • Wenjun Li,
  • Min Chen

摘要

As a remarkable model protein for studying excitation energy transfer (EET), Cryptophyte phycocyanin 645 (Cr-PC645) is not only important in the frontier research field on photosynthesis but also, like other kinds of phycocyanin, valuable in food and cosmetic industries as a kind of natural blue pigment protein. Here, we report a convenient preparation method for the purification of Cr-PC645 from cryptophytic algae, Chroomonas placoidea, mainly using hydrophobic interaction chromatography, and a study of EET characterization based on it. The final recovery of Cr-PC645 is 58.49% and the highest purity of final samples reaches 7.27 (A645/A280) in the obtained products. In addition to high purity, the properties of unchanged intact optical activity of purified Cr-PC645 are proved by comprehensively analyzing absorption, fluorescence, circular dichroism, and ultrafast time-resolved spectra. Furthermore, three time components of 6, 59, and 1021 ps are confirmed by 77 K time-resolved fluorescence spectroscopy experiments, which can be assigned to the time of energy transfer process between specific phycocyanobilins (PCBs), suggesting that the excitation energy is transferred from PCB-158 to PCB-82 in the same β subunit and localized the PCB-82 chromophores between different β subunits, then transferred to the downstream acceptors. In view of all these results, this optimized method is efficient and will be conducive to multi-field applications in the future.

Graphical Abstract