A Novel Recombinant Chlorophyllase1 from Chlamydomonas reinhardtii for the Production of Chlorophyllide Derivatives
A Novel Recombinant Chlorophyllase1 from Chlamydomonas reinhardtii for the Production of Chlorophyllide Derivatives
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DOI：
10.1021/acs.jafc.5b02787
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发表时间：
2015-11-04
影响因子：
6.1
通讯作者：
Shaw, Jei-Fu
中科院分区：
文献类型：
--
作者：
Chou, Yi-Li;Ko, Chia-Yun;Shaw, Jei-Fu
天然叶绿素代谢产物在体外表现出生理活性。在这项研究中，叶绿素酶1基因重组莱茵衣藻（CrCLH 1）的分离和鉴定。重组CrCLH 1可以进行叶绿素脱植基化并产生叶绿素和植醇。在瞬时测定中，确定CrCLH 1-绿色荧光蛋白的亚细胞定位在叶绿体外。生化分析表明，CrCLH 1的最适pH值为6.0，最适温度为40 ℃。酶动力学数据显示，重组CrCLH 1对叶绿素a的催化效率高于叶绿素B和细菌叶绿素a。根据高效液相色谱分析叶绿素水解，重组CrCLH 1催化叶绿素a的转化脱镁叶绿酸a在pH 5。因此，重组CrCLH 1可以用作生物催化剂来生产叶绿素衍生物。
Natural chlorophyll metabolites have exhibited physiological activity in vitro. In this study, a recombinant chlorophyllase1 gene from Chlamydomonas reinhardtii (CrCLH1) was isolated and characterized. Recombinant CrCLH1 can perform chlorophyll dephytylation and produce chlorophyllide and phytol. In a transient assay, the subcellular localization of CrCLH1-green fluorescent protein was determined to be outside the chloroplast. Biochemical analyses of the activity of recombinant CrCLH1 indicated that its optimal pH value and temperature are 6.0 and 40 degrees C, respectively. Enzyme kinetic data revealed that the recombinant CrCLH1 had a higher catalytic efficiency for chlorophyll a than for chlorophyll b and bacteriochlorophyll a. According to high-performance liquid chromatography analysis of chlorophyll hydrolysis, recombinant CrCLH1 catalyzed the conversion of chlorophyll a to pheophorbide a at pH 5. Therefore, recombinant CrCLH1 can be used as a biocatalyst to produce chlorophyllide derivatives.