title: Immobilization of Chlamydomonas reinhardtii CLH1 on APTES-Coated Magnetic Iron Oxide Nanoparticles and Its Potential in the Production of Chlorophyll Derivatives.
abstract: Recombinant Chlamydomonas reinhardtii chlorophyllase 1 (CrCLH1) that could catalyze chlorophyll hydrolysis to chlorophyllide and phytol in vitro was successfully expressed in Escherichia coli. The recombinant CrCLH1 was immobilized through covalent binding with a cubic (3-aminopropyl) triethoxysilane (APTES) coating on magnetic iron oxide nanoparticles (MIONPs), which led to markedly improved enzyme performance and decreased biocatalyst costs for potential industrial application. The immobilized enzyme exhibited a high immobilization yield (98.99 ± 0.91 mg/g of gel) and a chlorophyllase assay confirmed that the immobilized recombinant CrCLH1 retained enzymatic activity (722.3 ± 50.3 U/g of gel). Biochemical analysis of the immobilized enzyme, compared with the free enzyme, showed higher optimal pH and pH stability for chlorophyll-a hydrolysis in an acidic environment (pH 3-5). In addition, compared with the free enzyme, the immobilized enzyme showed higher activity in chlorophyll-a hydrolysis in a high temperature environment (50-60 °C). Moreover, the immobilized enzyme retained a residual activity of more than 64% of its initial enzyme activity after 14 cycles in a repeated-batch operation. Therefore, APTES-coated MIONP-immobilized recombinant CrCLH1 can be repeatedly used to lower costs and is potentially useful for the industrial production of chlorophyll derivatives.
authors.affiliation: Institute of Genomics and Bioinformatics, National Chung Hsing University, Taichung 40227, Taiwan. rover4rover4@gmail.com.|Agricultural Biotechnology Center, National Chung Hsing University, Taichung 40227, Taiwan. rover4rover4@gmail.com.
authors.forename: Chih-Chung
authors.initials: CC
authors.lastname: Yen
authors.affiliation: Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, Taiwan. ycchuang@nhri.org.tw.
authors.forename: Yao-Chen
authors.initials: YC
authors.lastname: Chuang
authors.affiliation: Institute of Plant and Microbial Biology, Academia Sinica, Taipei 11529, Taiwan. g874222@gate.sinica.edu.tw.
authors.forename: Chia-Yun
authors.initials: CY
authors.lastname: Ko
authors.affiliation: Institute of Plant and Microbial Biology, Academia Sinica, Taipei 11529, Taiwan. ochenlf@gate.sinica.edu.tw.
authors.forename: Long-Fang O
authors.initials: LF
authors.lastname: Chen
authors.affiliation: Department of Biological Science &amp; Technology, I-Shou University, Kaohsiung 840, Taiwan. cd710402@hotmail.com.
authors.forename: Sheau-Shyang
authors.initials: SS
authors.lastname: Chen
authors.affiliation: Department of Biological Science &amp; Technology, I-Shou University, Kaohsiung 840, Taiwan. didi0212.bce98g@g2.nctu.edu.tw.
authors.forename: Chia-Jung
authors.initials: CJ
authors.lastname: Lin
authors.affiliation: Department of Biological Science &amp; Technology, I-Shou University, Kaohsiung 840, Taiwan. qbowchou@gmail.com.
authors.forename: Yi-Li
authors.initials: YL
authors.lastname: Chou
authors.affiliation: Agricultural Biotechnology Center, National Chung Hsing University, Taichung 40227, Taiwan. shawjf@isu.edu.tw.|Department of Biological Science &amp; Technology, I-Shou University, Kaohsiung 840, Taiwan. shawjf@isu.edu.tw.
authors.forename: Jei-Fu
authors.initials: JF
authors.lastname: Shaw
mesh_terms: D020418:Algal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D002734:Chlorophyll / Q000737:chemistry*; D055590:Electromagnetic Phenomena; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005290:Ferric Compounds / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D053758:Nanoparticles / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D020418:Algal Proteins; D004800:Enzymes, Immobilized; D005290:Ferric Compounds; D011994:Recombinant Proteins; D002734:Chlorophyll; C000499:ferric oxide; D002265:Carboxylic Ester Hydrolases; C019875:chlorophyllase
keywords: Chlamydomonas reinhardtii chlorophyllase 1; chlorophyll derivatives; immobilized enzyme; magnetic iron oxide nanoparticles