Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - A solid-state NMR study of the immobilization of alpha-chymotrypsin on mesoporous silica
AU - Faure, Nicole E.
AU - Halling, Peter J.
AU - Wimperis, Stephen
PY - 2014/1/16
Y1 - 2014/1/16
N2 - Solid-state NMR spectroscopy was used to characterize a model biocatalyst system consisting of the enzyme alpha-chymotrypsin covalently immobilized on epoxide-silica ((glycidoxypropyl)trimethoxysilane, GOPS, grafted onto the surface of a silica gel). One- and two-dimensional H-1, C-13, and Si-29 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 in C-13 MAS NMR spectra recorded at B-o = 20 T. A number of further changes were observed in the C-13 and Si-29 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.
AB - Solid-state NMR spectroscopy was used to characterize a model biocatalyst system consisting of the enzyme alpha-chymotrypsin covalently immobilized on epoxide-silica ((glycidoxypropyl)trimethoxysilane, GOPS, grafted onto the surface of a silica gel). One- and two-dimensional H-1, C-13, and Si-29 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 in C-13 MAS NMR spectra recorded at B-o = 20 T. A number of further changes were observed in the C-13 and Si-29 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.
KW - NUCLEAR MAGNETIC-RESONANCE
KW - CROSS-POLARIZATION
KW - SUPPORT
KW - ENZYMES
KW - LIPASE
KW - HYDROLYSIS
KW - TITRATION
KW - PROTEINS
KW - FOAMS
KW - SI-29
U2 - 10.1021/jp4098414
DO - 10.1021/jp4098414
M3 - Journal article
VL - 118
SP - 1042
EP - 1048
JO - The Journal of Physical Chemistry C
JF - The Journal of Physical Chemistry C
SN - 1932-7447
IS - 2
ER -