Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds

Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds

In this study, silica-coated magnetic nanoparticles (SiMNPs) with isocyanatopropyltriethoxysilane as a metal-chelating ligand were prepared for the immobilization of His<sub>6</sub>-tagged <i>Escherichia coli</i> prolidase (His<sub>6</sub>-<i>Ec</i>Pep...

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Main Authors: Tzu-Fan Wang, Huei-Fen Lo, Meng-Chun Chi, Kuan-Ling Lai, Min-Guan Lin, Long-Liu Lin
Format: Article
Language:English
Published: MDPI AG 2019-07-01
Series:International Journal of Molecular Sciences
Subjects:
bacterial prolidase
magnetic nanoparticles
affinity immobilization
hydrolysis
diethyl/dimethyl paraoxon
Online Access:https://www.mdpi.com/1422-0067/20/15/3625
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spelling doaj-3590bd5d58e64b8fb5ea209b8d67b19b2020-11-25T02:29:28ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-07-012015362510.3390/ijms20153625ijms20153625Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus CompoundsTzu-Fan Wang0Huei-Fen Lo1Meng-Chun Chi2Kuan-Ling Lai3Min-Guan Lin4Long-Liu Lin5Department of Applied Chemistry, National Chiayi University, 300 Syuefu Road, Chiayi City 60004, TaiwanDepartment of Food Science and Technology, Hungkuang University, 1018 Taiwan Boulevard, Shalu District, Taichung City 43302, TaiwanDepartment of Applied Chemistry, National Chiayi University, 300 Syuefu Road, Chiayi City 60004, TaiwanDepartment of Applied Chemistry, National Chiayi University, 300 Syuefu Road, Chiayi City 60004, TaiwanInstitute of Molecular Biology, Academia Sinica, Nangang District, Taipei City 11529, TaiwanDepartment of Applied Chemistry, National Chiayi University, 300 Syuefu Road, Chiayi City 60004, TaiwanIn this study, silica-coated magnetic nanoparticles (SiMNPs) with isocyanatopropyltriethoxysilane as a metal-chelating ligand were prepared for the immobilization of His<sub>6</sub>-tagged <i>Escherichia coli</i> prolidase (His<sub>6</sub>-<i>Ec</i>PepQ). Under one-hour coupling, the enzyme-loading capacity for the Ni<sup>2+</sup>-functionalized SiMNPs (NiNTASiMNPs) was 1.5 mg/mg support, corresponding to about 58.6% recovery of the initial activity. Native and enzyme-bound NiNTASiMNPs were subsequently characterized by transmission electron microscopy (TEM), superparamagnetic analysis, X-ray diffraction, and Fourier transform infrared (FTIR) spectroscopy. As compared to free enzyme, His<sub>6</sub>-<i>Ec</i>PepQ@NiNTASiMNPs had significantly higher activity at 70 &#176;C and pH ranges of 5.5 to 10, and exhibited a greater stability during a storage period of 60 days and could be recycled 20 times with approximately 80% retention of the initial activity. The immobilized enzyme was further applied in the hydrolysis of two different organophosphorus compounds, dimethyl <i>p</i>-nitrophenyl phosphate (methyl paraoxon) and diethyl <i>p</i>-nitrophenyl phosphate (ethyl paraoxon). The experimental results showed that methyl paraoxon was a preferred substrate for His<sub>6</sub>-<i>Ec</i>PepQ and the kinetic behavior of free and immobilized enzymes towards this substance was obviously different. Taken together, the immobilization strategy surely provides an efficient means to deposit active enzymes onto NiNTASiMNPs for His<sub>6</sub>-<i>Ec</i>PepQ-mediated biocatalysis.https://www.mdpi.com/1422-0067/20/15/3625bacterial prolidasemagnetic nanoparticlesaffinity immobilizationhydrolysisdiethyl/dimethyl paraoxon
collection DOAJ
language English
format Article
sources DOAJ
author Tzu-Fan Wang
Huei-Fen Lo
Meng-Chun Chi
Kuan-Ling Lai
Min-Guan Lin
Long-Liu Lin
spellingShingle Tzu-Fan Wang
Huei-Fen Lo
Meng-Chun Chi
Kuan-Ling Lai
Min-Guan Lin
Long-Liu Lin
Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds
International Journal of Molecular Sciences
bacterial prolidase
magnetic nanoparticles
affinity immobilization
hydrolysis
diethyl/dimethyl paraoxon
author_facet Tzu-Fan Wang
Huei-Fen Lo
Meng-Chun Chi
Kuan-Ling Lai
Min-Guan Lin
Long-Liu Lin
author_sort Tzu-Fan Wang
title Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds
title_short Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds
title_full Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds
title_fullStr Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds
title_full_unstemmed Affinity Immobilization of a Bacterial Prolidase onto Metal-Ion-Chelated Magnetic Nanoparticles for the Hydrolysis of Organophosphorus Compounds
title_sort affinity immobilization of a bacterial prolidase onto metal-ion-chelated magnetic nanoparticles for the hydrolysis of organophosphorus compounds
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1422-0067
publishDate 2019-07-01
description In this study, silica-coated magnetic nanoparticles (SiMNPs) with isocyanatopropyltriethoxysilane as a metal-chelating ligand were prepared for the immobilization of His<sub>6</sub>-tagged <i>Escherichia coli</i> prolidase (His<sub>6</sub>-<i>Ec</i>PepQ). Under one-hour coupling, the enzyme-loading capacity for the Ni<sup>2+</sup>-functionalized SiMNPs (NiNTASiMNPs) was 1.5 mg/mg support, corresponding to about 58.6% recovery of the initial activity. Native and enzyme-bound NiNTASiMNPs were subsequently characterized by transmission electron microscopy (TEM), superparamagnetic analysis, X-ray diffraction, and Fourier transform infrared (FTIR) spectroscopy. As compared to free enzyme, His<sub>6</sub>-<i>Ec</i>PepQ@NiNTASiMNPs had significantly higher activity at 70 &#176;C and pH ranges of 5.5 to 10, and exhibited a greater stability during a storage period of 60 days and could be recycled 20 times with approximately 80% retention of the initial activity. The immobilized enzyme was further applied in the hydrolysis of two different organophosphorus compounds, dimethyl <i>p</i>-nitrophenyl phosphate (methyl paraoxon) and diethyl <i>p</i>-nitrophenyl phosphate (ethyl paraoxon). The experimental results showed that methyl paraoxon was a preferred substrate for His<sub>6</sub>-<i>Ec</i>PepQ and the kinetic behavior of free and immobilized enzymes towards this substance was obviously different. Taken together, the immobilization strategy surely provides an efficient means to deposit active enzymes onto NiNTASiMNPs for His<sub>6</sub>-<i>Ec</i>PepQ-mediated biocatalysis.
topic bacterial prolidase
magnetic nanoparticles
affinity immobilization
hydrolysis
diethyl/dimethyl paraoxon
url https://www.mdpi.com/1422-0067/20/15/3625
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