Immobilization of <i>β</i>-glucuronidase in lysozyme-induced biosilica particles to improve its stability

doi:10.1007/s11705-014-1421-2

Front. Chem. Sci. Eng.

2014, Vol. 8

Issue (3) : 353-361 https://doi.org/10.1007/s11705-014-1421-2

RESEARCH ARTICLE

Immobilization of β-glucuronidase in lysozyme-induced biosilica particles to improve its stability

Xiaokai SONG¹,Zhongyi JIANG^1,²,Lin LI¹,Hong WU^1,^2,^*(

)

1. Key Laboratory for Green Chemical Technology of Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
2. Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China

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Abstract

Mesoporous silica particles were prepared for efficient immobilization of the β-glucuronidase (GUS) through a biomimetic mineralization process, in which the solution containing lysozyme and GUS were added into the prehydrolyzed tetraethoxysilane (TEOS) solution. The silica particles were formed in a way of biomineralization under the catalysis of lysozyme and GUS was immobilized into the silica particles simultaneously during the precipitation process. The average diameter of the silica particles is about 200 nm with a pore size of about 4 nm. All the enzyme molecules are tightly entrapped inside the biosilica nanoparticles without any leaching even under a high ionic strength condition. The immobilized GUS exhibits significantly higher thermal and pH stability as well as the storage and recycling stability compared with GUS in free form. No loss in the enzyme activity of the immobilized GUS was found after 30-day’s storage, and the initial activity could be well retained after 12 repeated cycles.

Keywords storage and recycling stability silica nanoparticles biocatalysis biomimetic synthesis β-glucuronidase encapsulation

Corresponding Author(s): Hong WU

Issue Date: 11 October 2014

Cite this article:

Xiaokai SONG,Zhongyi JIANG,Lin LI, et al. Immobilization of β-glucuronidase in lysozyme-induced biosilica particles to improve its stability[J]. Front. Chem. Sci. Eng., 2014, 8(3): 353-361.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-014-1421-2
https://academic.hep.com.cn/fcse/EN/Y2014/V8/I3/353

Fig.1 TEM images of (a) the pristine lysozyme-induced biosilica particles and (b) those with GUS inside

Fig.2 N₂ adsorption-desorption isotherm and pore size distribution of the biosilica particles

Fig.3 ²⁹Si NMR of the lysozyme-induced biosilica

Fig.4 Scheme 1 Schematic view of the immobilization process of GUS in biosilica particles

Fig.5 Effect of (a) temperature and (b) pH on the activity of free and immobilized GUSs

Fig.6 (a) Thermal and (b) pH stability of free and immobilized GUSs

Fig.7 Productivity of baicalein with reaction time

Tab.1 Kinetic parameters for free and immobilized GUS

Fig.8 Storage stability of free and immobilized GUSs

Fig.9 Recycling stability of immobilized GUS

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