生物学杂志

生物学杂志

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重组LHCⅡ与叶绿素a的体外结合及其光电性能表征

  

  1. 1. 青岛大学 生命科学学院, 青岛 266071; 2.中国石油大学（华东）生物工程与技术中心, 青岛 266555;3. 中国科学院 烟台海岸带研究所, 烟台 264003
  • 出版日期:2018-08-18 发布日期:2018-08-18
  • 通讯作者: 葛保胜，博士，副教授，从事微藻能源及综合利用技术研究,E-mail：gebaosheng@upc.edu.cn
  • 作者简介:生，主要研究方向为微生物工程,E-mail：houqq327@163.com
  • 基金资助:
    国家重点研发计划项目（2016YFE0106700）；国家自然科学基金项目（41176144）

In vitro reconstitution of LHCⅡ recombinant with chlorophyll-a and characterization of its photoelectric properties

  1. 1. College of Life Sciences, Qingdao University, Qingdao 266071; 2. Center for Bioengineering and  Biotechnology, China University of Petroleum(East China), Qingdao 266555; 3. Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China
  • Online:2018-08-18 Published:2018-08-18

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摘要： 通过克隆莱茵衣藻的lhcⅡ基因，构建pET-28a-lhcⅡ质粒，在E.coli BL21中进行表达、纯化，获得重组蛋白LHCⅡ，将其在体外与叶绿素a进行结合，通过调整叶绿素a与LHCⅡ的摩尔比，获得叶绿素a与LHCⅡ结合率的饱和曲线，并将其和相同浓度的叶绿素a分别固定在染料敏化TiO2电极上，对比两者的光电转换性能。通过分析可得，重组表达的LHCⅡ在体外可以结合8个叶绿素a分子。在AM1.5，光照强度为100 mW/cm2的条件下，测得重组后LHCⅡ复合物敏化电池的短路电流ISC为0.493 mA/cm2，开路电压VOC为0.491 V，填充因子FF为0.679，效率η为0.165%；相同浓度下,叶绿素a敏化电池的短路电流ISC为0.596 mA/cm2，开路电压VOC为0.482 V，填充因子FF为0.676，效率η为0.194%。结果显示，重组LHCⅡ蛋白可以在体外与叶绿素a结合，其结合活性良好。重建之后的LHCⅡ色素-蛋白复合物可以在染料敏化太阳能电池系统中进行完整的电子传递，在低浓度下，表现出良好的光电性能。为大规模制备LHCⅡ、研究重组蛋白LHCⅡ在体外与叶绿素的结合活性及其在光电转换方面的应用奠定了基础。

关键词: LHCⅡ, 叶绿素, 染料敏化太阳能电池

Abstract: The lhcⅡ gene from Chlamydomonas reinhardtii was cloned and expressed in Escherichia coli. The LHCⅡrecombinant was prepared using genetic engineering technology, and then reconstructed with chlorophyll a in vitro. The saturation curve of the complex of LHCⅡ/chlorophyll-a was obtained. The free chlorophyll a and the complexs of LHCⅡ/chlorophyll a (at same concentration) were fixed on the TiO2 electrode, and the photoelectric conversion properties of these two materials were compared. One recombinant LHCⅡ bound with 8 chlorophyll a molecules. Under the conditions of AM1.5, light intensity of 100 mW/cm2, shortcut circuit current of LHCⅡ/chlorophyll-a complex is 0.493 mA/cm2, open circuit voltage is 0.491 V, fill-factor is 0.679, and the efficiency is 0.165%. In comparison, shortcut circuit current of free chlorophyll a is 0.596 mA/cm2, open circuit voltage is 0.482 V, and fill-factor is 0.676, efficiency is 0.194%. The recombinant LHCⅡ protein obtained from E. coli BL21 was successfully combined with chlorophyll a in vitro with similar binding activity as its native proteins. The reconstituted pigment-protein complex fulfilled the light harvesting and electron transport functions in a dye-sensitized solar cell system, and showed promising photoelectric properties at lower concentrations. In this study, large-scale preparation of LHCⅡ, the binding activity of recombinant protein LHCⅡ with chlorophyll in vitro, and its application in photoelectric conversion has laid a foundation.

Key words: LHCⅡ, chlorophylls, dye-sensitized solar cells