Enhanced activity and stability of lysozyme by immobilization in the matching nanochannels of mesoporous silica nanoparticles

Kun Che Kao, Tien Sung Lin, Chung Yuan Mou

研究成果: 雜誌貢獻文章同行評審

83 引文 斯高帕斯(Scopus)

摘要

It is highly desirable to study the kinetics and spectroscopy of enzymes in a crowded and controllable microenvironment. In this work, we employ mesoporous silica of matching pore sizes to confine lysozyme in order to mimic enzyme in a crowded environment. The stability and activity of lysozyme immobilized in mesoporous silica nanoparticle (MSN) of various pore sizes were studied and correlated to spectroscopic data of the immobilized enzyme. By site-selective surface functionalization, we were able to avoid protein adsorbing on the external surfaces of MSNs and study specifically the protein immobilized in the nanochannels. Solution spectroscopic methods, CD and fluorescence, were used to study the secondary and tertiary structures of the immobilized enzyme because MSNs could be suspended very well in solution. To study the catalytic activity of lysozyme, we employed 4-methylumbelliferyl β-d-N,N,N′- triacetylchitotrioside as a substrate that was hydrolyzed and detected by fluorescence spectroscopy. 8-Anilino-1-naphthalenesulfonic acid was utilized as a fluorescence probe to characterize the protein-binding site. The conformation, thermal stability, and catalytic activity of lysozyme were sensitive to the curvature of the silica materials. The activity of the lysozyme immobilized in the 5.6 nm mesopores of MSNs was higher than those of native enzymes. The enhanced activity was attributed to subtle change in tertiary structure of lysozyme in the crowded microenvironment in the mesopores.

原文英語
頁(從 - 到)6734-6743
頁數10
期刊Journal of Physical Chemistry C
118
發行號13
DOIs
出版狀態已發佈 - 4月 3 2014
對外發佈

ASJC Scopus subject areas

  • 電子、光磁材料
  • 能源(全部)
  • 物理與理論化學
  • 表面、塗料和薄膜

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