Immobilization of Chlamydomonas reinhardtii CLH1 on APTES-Coated Magnetic Iron Oxide Nanoparticles and Its Potential in the Production of Chlorophyll Derivatives

Molecules

Chia-Jung Lin¹  Jei-Fu Shaw¹  Yi-Li Chou²  Sheau-Shyang Chen²  Yao-Chen Chuang³  Chih-Chung Yen⁴  Long-Fang O. Chen⁵  Chia-Yun Ko⁵ 

[1] Technology, I-Shou University, Kaohsiung 840, Taiwan;;Department of Biological Science &Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli 35053, Taiwan;Institute of Genomics and Bioinformatics, National Chung Hsing University, Taichung 40227, Taiwan;Institute of Plant and Microbial Biology, Academia Sinica, Taipei 11529, Taiwan;

关键词: Chlamydomonas reinhardtii chlorophyllase 1;    immobilized enzyme;    magnetic iron oxide nanoparticles;    chlorophyll derivatives;   

DOI  :  10.3390/molecules21080972

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown