A solid-state NMR study of the immobilization of alpha-chymotrypsin on mesoporous silica

N. E. Faure, P. J. Halling, S. Wimperis

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

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-dimensional 1H, 13C, and 29Si 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 13C MAS NMR spectra recorded at B0 = 20 T. A number of further changes were observed in the 13C and 29Si 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.
LanguageEnglish
Pages1042-1048
Number of pages7
JournalJournal of Physical Chemistry C
Volume118
Issue number2
Early online date20 Dec 2013
DOIs
Publication statusPublished - 16 Jan 2014

Fingerprint

immobilization
Silicon Dioxide
Magic angle spinning
Epoxy Compounds
Silica
Nuclear magnetic resonance
epoxy compounds
silicon dioxide
solid state
nuclear magnetic resonance
Biocatalysts
metal spinning
Enzymes
Carbon
enzymes
Immobilized Enzymes
Silica Gel
Silica gel
Silicon
support systems

Keywords

  • solid state NMR spectroscopy
  • biocatalyst system
  • biocatalyst system modeling

Cite this

@article{140cd37af4c94454ae1a5cbb5e9de6bf,
title = "A solid-state NMR study of the immobilization of alpha-chymotrypsin on mesoporous silica",
abstract = "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-dimensional 1H, 13C, and 29Si 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 13C MAS NMR spectra recorded at B0 = 20 T. A number of further changes were observed in the 13C and 29Si 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.",
keywords = "solid state NMR spectroscopy, biocatalyst system, biocatalyst system modeling",
author = "Faure, {N. E.} and Halling, {P. J.} and S. Wimperis",
note = "This document is the unedited Author’s version of a Submitted Work that was subsequently accepted for publication in Journal of Physical Chemistry C, copyright {\circledC} American Chemical Society after peer review. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/jp4098414.",
year = "2014",
month = "1",
day = "16",
doi = "10.1021/jp4098414",
language = "English",
volume = "118",
pages = "1042--1048",
journal = "Journal of Physical Chemistry C",
issn = "1932-7447",
publisher = "American Chemical Society",
number = "2",

}

A solid-state NMR study of the immobilization of alpha-chymotrypsin on mesoporous silica. / Faure, N. E.; Halling, P. J.; Wimperis, S.

In: Journal of Physical Chemistry C, Vol. 118, No. 2, 16.01.2014, p. 1042-1048.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A solid-state NMR study of the immobilization of alpha-chymotrypsin on mesoporous silica

AU - Faure, N. E.

AU - Halling, P. J.

AU - Wimperis, S.

N1 - This document is the unedited Author’s version of a Submitted Work that was subsequently accepted for publication in Journal of Physical Chemistry C, copyright © American Chemical Society after peer review. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/jp4098414.

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 α-chymotrypsin covalently immobilized on epoxide-silica ((glycidoxypropyl)trimethoxysilane, GOPS, grafted onto the surface of a silica gel). One- and two-dimensional 1H, 13C, and 29Si 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 13C MAS NMR spectra recorded at B0 = 20 T. A number of further changes were observed in the 13C and 29Si 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 α-chymotrypsin covalently immobilized on epoxide-silica ((glycidoxypropyl)trimethoxysilane, GOPS, grafted onto the surface of a silica gel). One- and two-dimensional 1H, 13C, and 29Si 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 13C MAS NMR spectra recorded at B0 = 20 T. A number of further changes were observed in the 13C and 29Si 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 - solid state NMR spectroscopy

KW - biocatalyst system

KW - biocatalyst system modeling

UR - http://pubs.acs.org/journal/jpccck

U2 - 10.1021/jp4098414

DO - 10.1021/jp4098414

M3 - Article

VL - 118

SP - 1042

EP - 1048

JO - Journal of Physical Chemistry C

T2 - Journal of Physical Chemistry C

JF - Journal of Physical Chemistry C

SN - 1932-7447

IS - 2

ER -