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

Overview

Journal J Agric Food Chem

Specialties Chemistry
Nutritional Sciences

Date 2015 Oct 20

PMID 26478543

Citations 9

Authors

Yi-Li Chou

Chia-Yun Ko

Chih-Chung Yen

Long-Fang O Chen

Jei-Fu Shaw

Affiliations

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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.

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