A Solid-State NMR Study of the Immobilization of a-Chymotrypsin on Mesoporous Silica
A Solid-State NMR Study of the Immobilization of a-Chymotrypsin on Mesoporous Silica
复制标题
介孔二氧化硅固定化α-胰凝乳蛋白酶的固态核磁共振研究
DOI：
10.1021/jp4098414
复制
发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Fauré N
中科院分区：
文献类型：
--
作者：
Fauré N
固态核磁共振波谱用于表征模型生物催化剂系统，该系统由共价固定在环氧化物二氧化硅（（缩水甘油氧基丙基）三甲氧基硅烷，GOPS，接枝到硅胶表面）上的α-胰凝乳蛋白酶组成。采用一维和二维 1 H、 13 C 和 29 Si 魔角旋转（MAS）NMR 技术。首先对支撑系统（环氧化物-二氧化硅）进行表征，并且可以在二氧化硅和 GOPS 连接体中指定硅和碳物种。蛋白质附着后，固定化酶中的羰基碳 (175 ppm) 在 B0 = 20 T 记录的 13 C MAS NMR 光谱中可见。在固定化过程中，在 13 C 和 29 Si MAS NMR 光谱中观察到许多进一步的变化，这是由表面二氧化硅物质的交联和通过亲核攻击打开环氧化物官能团引起的。这项研究显示了多核固态核磁共振在分子水平上更好地了解固体生物催化剂系统的潜力。
Solid-state NMR spectroscopy was used to characterize a model biocatalyst system consisting of the enzyme α-chymotrypsin covalently immobilized on epoxide-silica ((glycidoxypropyl)trimethoxysilane, GOPS, grafted onto the surface of a silica gel). One- and two-dimensional1H,13C, and29Si magic angle spinning (MAS) NMR techniques were employed. The support system (epoxide-silica) was characterized first and it was possible to assign silicon and carbon species in both the silica and the GOPS linker. After attachment of the protein, carbonyl carbons (175 ppm) in the immobilized enzyme were visible in13C MAS NMR spectra recorded atB0= 20 T. A number of further changes were observed in the13C and29Si MAS NMR spectra during the immobilization process, arising from a cross-linking of the surface silica species and an opening of the epoxide functional group by nucleophilic attack. This study shows the potential of multinuclear solid-state NMR for obtaining a better understanding of solid biocatalyst systems at the molecular level.