title: A Novel Recombinant Chlorophyllase1 from Chlamydomonas reinhardtii for the Production of Chlorophyllide Derivatives.
abstract: 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 °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.
authors.affiliation: Department of Biological Science and Technology, I-Shou University , Kaohsiung 82445, Taiwan.
authors.forename: Yi-Li
authors.initials: YL
authors.lastname: Chou
authors.affiliation: Institute of Plant and Microbial Biology, Academia Sinica , Taipei 11529, Taiwan.
authors.forename: Chia-Yun
authors.initials: CY
authors.lastname: Ko
authors.affiliation: Institute of Genomics and Bioinformatics, National Chung Hsing University , Taichung 40227, Taiwan.|Agricultural Biotechnology Center, National Chung Hsing University , Taichung 40227, Taiwan.
authors.forename: Chih-Chung
authors.initials: CC
authors.lastname: Yen
authors.affiliation: Institute of Plant and Microbial Biology, Academia Sinica , Taipei 11529, Taiwan.
authors.forename: Long-Fang O
authors.initials: LF
authors.lastname: Chen
authors.affiliation: Department of Biological Science and Technology, I-Shou University , Kaohsiung 82445, Taiwan.|Agricultural Biotechnology Center, National Chung Hsing University , Taichung 40227, Taiwan.
authors.forename: Jei-Fu
authors.initials: JF
authors.lastname: Shaw
mesh_terms: D055162:Biocatalysis; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D002735:Chlorophyllides / Q000737:chemistry*; D007700:Kinetics; D015394:Molecular Structure
chemical_list: D002735:Chlorophyllides; D002265:Carboxylic Ester Hydrolases; C019875:chlorophyllase
keywords: Chlamydomonas reinhardtii; chlorophyllase; chlorophyllide derivatives; enzyme kinetics