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Enzyme Microb Technol. 2019 Mar;122:26-35. doi: 10.1016/j.enzmictec.2018.12.007. Epub 2018 Dec 13.

Horseradish peroxidase immobilized on the magnetic composite microspheres for high catalytic ability and operational stability.

Author information

1
School of Pharmaceutical Science and Technology, Suzhou Chien-Shiung Institute of Technology, Taicang, 215400, China.
2
School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China.
3
School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China. Electronic address: hanjuan@ujs.edu.cn.
4
State Key Laboratory of Food Additive and Condiment Testing, Zhenjiang Customs District P.R. of China, Zhenjiang, 212008, China. Electronic address: chentong2006@163.com.
5
School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013, China.

Abstract

In the present study, a novel and efficient immobilization for horseradish peroxidase (HRP) had been developed by using 6-arm magnetic composite microsphere (Fe3O4@PAA-6-arm-PEG-NH2) containing 6-arm polyethylene glycol (6-arm-PEG-NH2) and Fe3O4. The morphology and chemical properties of Fe3O4@PAA-6-arm-PEG-NH2 were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectra (FTIR), X-ray powder diffraction (XRD), vibrating sample magnetometer (VSM) and thermogravimetric analysis (TGA). The immobilization conditions and loading capacity of the carrier toward HRP were also investigated. According to the results, the optimum immobilization conditions were as follows: glutaraldehyde concentration of 0.10 mol L-1, the carrier concentration of 7 mg L-1, the temperature of 35 °C and immobilization time of 180 min. The loading capacity of the immobilized HRP arrived to 139.82 mg g-1 (RSD = 0.87%). Compared with free HRP, the stability of the immobilized HRP had been enhanced in a wide range of temperature and pH. The activity of the immobilized HRP retained 71.05% of its initial activity after 60 days storage. It was also found that the activity of the immobilized HRP retained 61.06% of its initial activity after eight times of successive reuse. The immobilized HRP could hydrolyze phenol to 94.4% within 10 min. It proved that the immobilized HRP could improve the performance of HRP, which had a good prospect in biological application.

KEYWORDS:

Enzymatic activity; Horseradish peroxidase; Multi-arm magnetic composite; Phenol degradation

PMID:
30638506
DOI:
10.1016/j.enzmictec.2018.12.007
[Indexed for MEDLINE]

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