Tyrosinase immobilization on aminated magnetic nanoparticles by physical adsorption combined with covalent crosslinking with improved catalytic activity, reusability and storage stability

被引:75
作者
Liu, Dong-Mei [1 ,2 ,3 ]
Chen, Juan [1 ,2 ]
Shi, Yan-Ping [1 ,2 ]
机构
[1] Chinese Acad Sci, Lanzhou Inst Chem Phys, CAS Key Lab Chem Northwestern Plant Resources, Lanzhou 730000, Gansu, Peoples R China
[2] Chinese Acad Sci, Lanzhou Inst Chem Phys, Key Lab Nat Med Gansu Prov, Lanzhou 730000, Gansu, Peoples R China
[3] Univ Chinese Acad Sci, Chinese Acad Sci, Beijing 100049, Peoples R China
基金
中国国家自然科学基金;
关键词
Tyrosinase; Aminated magnetic nanoparticles; Physical adsorption; Covalent crosslinking; Inhibitor screening; CANDIDA-RUGOSA LIPASE; GOLD NANOPARTICLES; PROTEIN ADSORPTION; CHITOSAN BEADS; SOL-GEL; BIOSENSOR; PERFORMANCE; ENZYME; CHROMATOGRAPHY; BIOCATALYSIS;
D O I
10.1016/j.aca.2017.12.022
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
In the present study, the immobilization method of physical adsorption combined with covalent crosslinking was developed to avoid the shortcomings of both the noncovalent and covalent coupling methods. For the first time, tyrosinase (TYR) was immobilized on the surface of aminated magnetic nanoparticles (Fe3O4-NH2) by the developed method. TYR was firstly adsorbed on the surface of Fe3O4-NH2 by electrostatic interaction, and then by covalent crosslinking with glutaraldehyde (GA), TYR was firmly immobilized on the supports. The immobilized TYR showed enhanced pH and temperature endurances at the optimum pH of 7.0 and temperature of 35 degrees C. TYR reusability was significantly improved after immobilization and it retained 61.4 +/- 2.3% of its initial activity after 5 repeated cycles. Immobilized TYR also showed improved storage stability (73.2 +/- 1.1% after 30 days of storage at 4 degrees C). In addition, the immobilized TYR showed a higher biological affinity to substrate owing to the stabilization of TYR in its active conformation by electrostatic interaction prior to covalent crosslinking. Finally, the immobilized TYR was used to screen of enzyme inhibitors from 11 traditional Chinese medicines (TCMs) to validate whether this method can be used for enzyme inhibitor screening or not. (c) 2018 Elsevier B.V. All rights reserved.
引用
收藏
页码:90 / 98
页数:9
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