Optimum extraction and energy transfer dynamics in Phycocyanin 620 from Thermosynechococcus vulcanus

IF 4.2 3区工程技术 Q2 CHEMISTRY, APPLIED

Dyes and Pigments Pub Date : 2025-05-27 DOI:10.1016/j.dyepig.2025.112919

Yang Pu , Xiangyu Lan , Jiayu Wang , Chuanxu Liu , Juan Ding , Wenjun Li

{"title":"Optimum extraction and energy transfer dynamics in Phycocyanin 620 from Thermosynechococcus vulcanus","authors":"Yang Pu , Xiangyu Lan , Jiayu Wang , Chuanxu Liu , Juan Ding , Wenjun Li","doi":"10.1016/j.dyepig.2025.112919","DOIUrl":null,"url":null,"abstract":"<div><div>Phycocyanin 620 (PC620) is a water-soluble blue phycobiliprotein (PBP) originating from hot spring cyanobacteria — Thermosynechococcus vulcanus (Tv). It has been recognized as a valuable material with great potential for application to theoretical studies and commercial products. So, in our work, the PC620 was purified by a minimalistic procedure through hydrophobic chromatography and used for the study of excitation energy transfer (EET) characterization. The results show that the obtained PC620 with a final recovery of 81.5 % and an analytical grade purity at 5.21 (A620/A280) was reached through the process that consisted of liquid nitrogen grinding, 20 %–50 % ammonium sulfate fractionation and one-step hydrophobic interaction chromatography. The final PC620 showed a maximum absorption peak at 620 nm, a fluorescence emission peak at 652 nm and a positive peak of circular dichroism (CD) spectra in the visible range at 626 nm. Then, joint analysis of absorption spectra, fluorescence spectra, and CD spectra confirmed that the structural and functional integrity of the samples was retained compared with natural PC620. In addition, the exciton coupling in PC620 has been excluded by CD measurements. Furthermore, in an accurate experimental determination of the EET lifetime in PC620, two energy migration pathways from α84 to β84 in a period of ∼0.25 ps and then from β155 to β84 in a period of ∼5 ps were directly resolved by ultrafast time-resolved spectroscopy. The calculated results indicated that the EET process in PC620 is dominated by the Förster resonance energy transfer (FRET) mechanism. This work should be conducive to the highly efficient preparation of the analytical grade PC620 using a substantially simpler purification method for exploring the applications of theoretical research and the commercial field.</div></div>","PeriodicalId":302,"journal":{"name":"Dyes and Pigments","volume":"241 ","pages":"Article 112919"},"PeriodicalIF":4.2000,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Dyes and Pigments","FirstCategoryId":"88","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S014372082500289X","RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, APPLIED","Score":null,"Total":0}

引用次数: 0

Abstract

Phycocyanin 620 (PC₆₂₀) is a water-soluble blue phycobiliprotein (PBP) originating from hot spring cyanobacteria — Thermosynechococcus vulcanus (Tv). It has been recognized as a valuable material with great potential for application to theoretical studies and commercial products. So, in our work, the PC₆₂₀ was purified by a minimalistic procedure through hydrophobic chromatography and used for the study of excitation energy transfer (EET) characterization. The results show that the obtained PC₆₂₀ with a final recovery of 81.5 % and an analytical grade purity at 5.21 (A₆₂₀/A₂₈₀) was reached through the process that consisted of liquid nitrogen grinding, 20 %–50 % ammonium sulfate fractionation and one-step hydrophobic interaction chromatography. The final PC₆₂₀ showed a maximum absorption peak at 620 nm, a fluorescence emission peak at 652 nm and a positive peak of circular dichroism (CD) spectra in the visible range at 626 nm. Then, joint analysis of absorption spectra, fluorescence spectra, and CD spectra confirmed that the structural and functional integrity of the samples was retained compared with natural PC₆₂₀. In addition, the exciton coupling in PC₆₂₀ has been excluded by CD measurements. Furthermore, in an accurate experimental determination of the EET lifetime in PC₆₂₀, two energy migration pathways from α84 to β84 in a period of ∼0.25 ps and then from β155 to β84 in a period of ∼5 ps were directly resolved by ultrafast time-resolved spectroscopy. The calculated results indicated that the EET process in PC₆₂₀ is dominated by the Förster resonance energy transfer (FRET) mechanism. This work should be conducive to the highly efficient preparation of the analytical grade PC₆₂₀ using a substantially simpler purification method for exploring the applications of theoretical research and the commercial field.

Abstract Image

查看原文本刊更多论文

vulcanhot synechococcus藻蓝蛋白620的最佳提取及能量传递动力学

藻蓝蛋白620 （Phycocyanin 620, PC620）是一种水溶性蓝色藻胆蛋白（PBP），来源于温泉蓝藻——vulcanus Thermosynechococcus （Tv）。它被认为是一种有价值的材料，在理论研究和商业产品中具有很大的应用潜力。因此，在我们的工作中，通过疏水色谱法纯化PC620，并将其用于激发能转移（EET）表征的研究。结果表明，通过液氮研磨、20% - 50%硫酸铵分馏和一步疏水相互作用层析，所得PC620最终回收率为81.5%，分析级纯度为5.21 （A620/A280）。最终的PC620在620nm处有最大吸收峰，在652nm处有荧光发射峰，在626nm处有圆二色（CD）光谱的正峰。通过对吸收光谱、荧光光谱和CD光谱的联合分析，证实与天然PC620相比，样品保持了结构和功能的完整性。此外，通过CD测量排除了PC620中的激子耦合。此外，在精确测定PC620中EET寿命的实验中，通过超快时间分辨光谱直接分辨了α84到β84在~ 0.25 ps周期内和β155到β84在~ 5 ps周期内的两条能量迁移路径。计算结果表明，PC620中的EET过程以Förster共振能量传递（FRET）机制为主。本工作将有助于以更简单的纯化方法高效制备分析级PC620，探索其在理论研究和商业领域的应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Dyes and Pigments 工程技术-材料科学：纺织

CiteScore

8.20

自引率

13.30%

发文量

933

审稿时长

33 days

期刊介绍： Dyes and Pigments covers the scientific and technical aspects of the chemistry and physics of dyes, pigments and their intermediates. Emphasis is placed on the properties of the colouring matters themselves rather than on their applications or the system in which they may be applied. Thus the journal accepts research and review papers on the synthesis of dyes, pigments and intermediates, their physical or chemical properties, e.g. spectroscopic, surface, solution or solid state characteristics, the physical aspects of their preparation, e.g. precipitation, nucleation and growth, crystal formation, liquid crystalline characteristics, their photochemical, ecological or biological properties and the relationship between colour and chemical constitution. However, papers are considered which deal with the more fundamental aspects of colourant application and of the interactions of colourants with substrates or media. The journal will interest a wide variety of workers in a range of disciplines whose work involves dyes, pigments and their intermediates, and provides a platform for investigators with common interests but diverse fields of activity such as cosmetics, reprographics, dye and pigment synthesis, medical research, polymers, etc.