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

J Agric Food Chem. 2015 Nov 4;63(43):9496-503. doi: 10.1021/acs.jafc.5b02787. Epub 2015 Oct 26.

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.

Keywords: Chlamydomonas reinhardtii; chlorophyllase; chlorophyllide derivatives; enzyme kinetics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocatalysis
  • Carboxylic Ester Hydrolases / chemistry*
  • Carboxylic Ester Hydrolases / genetics
  • Carboxylic Ester Hydrolases / metabolism
  • Chlamydomonas reinhardtii / chemistry
  • Chlamydomonas reinhardtii / enzymology*
  • Chlamydomonas reinhardtii / genetics
  • Chlorophyllides / chemistry*
  • Kinetics
  • Molecular Structure

Substances

  • Chlorophyllides
  • Carboxylic Ester Hydrolases
  • chlorophyllase