A Novel Recombinant Chlorophyllase1 from Chlamydomonas reinhardtii for the Production of Chlorophyllide Derivatives

被引：15
|
作者：
Chou, Yi-Li [1 ]
Ko, Chia-Yun [2 ]
Yen, Chih-Chung [3 ,4 ]
Chen, Long-Fang O. [2 ]
Shaw, Jei-Fu [1 ,4 ]
机构：
[1] I Shou Univ, Dept Biol Sci & Technol, Kaohsiung 82445, Taiwan
[2] Acad Sinica, Inst Plant & Microbial Biol, Taipei 11529, Taiwan
[3] Natl Chung Hsing Univ, Inst Genom & Bioinformat, Taichung 40227, Taiwan
[4] Natl Chung Hsing Univ, Agr Biotechnol Ctr, Taichung 40227, Taiwan
来源：
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2015年 / 63卷 / 43期
关键词：
Chlamydomonas reinhardtii; chlorophyllase; chlorophyllide derivatives; enzyme kinetics; LEAF SENESCENCE; CHLORELLA-PROTOTHECOIDES; DEGRADATION; BREAKDOWN; PURIFICATION; HYDROLASE; ENZYME; PHEOPHORBIDE; BIOSYNTHESIS; CLONING;
D O I：
10.1021/acs.jafc.5b02787
中图分类号：
S [农业科学];
学科分类号：
09 ;
摘要：
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.
引用
收藏
页码：9496 / 9503
页数：8
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