Abstract
Nuclear magnetic resonance (NMR) is a powerful tool for investigating atomic-scale structure in heterogeneous or composite materials where long-range order is absent. In this work solid-state 1H and 1H-detected NMR experiments were performed with fast magic angle spinning (νR = 75 kHz) and at high magnetic fields (B0 = 20 T) and used to gain structural insight into a heterogeneous biocatalyst consisting of an enzyme, human carbonic anhydrase II (hCA II), covalently immobilized on epoxy-functionalized silica. Two-dimensional 1H-1H NOESY-type correlation experiments were able to provide information on 1H environments in silica, epoxy-silica and the immobilized enzyme. Two distinct signals originating from water protons were observed: water associated with the surface of the silica and the water associated with the immobilized enzyme. Additional two-dimensional 1H-1H double–single quantum (DQ-SQ) correlation experiments suggested that the immobilized enzyme is not in close contact with the silica surface. Most significantly, comparison of two-dimensional 1H-15N spectra of the immobilized enzyme and the solution-state enzyme confirmed that the structural integrity of the protein is well preserved upon covalent immobilization.
| Language | English |
|---|---|
| Pages | 7-11 |
| Number of pages | 5 |
| Journal | Solid State Nuclear Magnetic Resonance |
| Volume | 92 |
| Early online date | 14 Mar 2018 |
| DOIs | |
| Publication status | E-pub ahead of print - 14 Mar 2018 |
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Keywords
- solid-state NMR
- biocatalysis
- covalent immobilization
- epoxy-functionalized silica
- hCA II
- heterogeneous biocatalysts
- human carbonic anhydrase II
- immobilized enzymes
Cite this
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Two-dimensional 1H and 1H-detected NMR study of a heterogeneous biocatalyst using fast MAS at high magnetic fields. / Varghese, Sabu; Halling, Peter J.; Häussinger, Daniel; Wimperis, Stephen.
Vol. 92, 14.03.2018, p. 7-11.Research output: Contribution to journal › Article
TY - JOUR
T1 - Two-dimensional 1H and 1H-detected NMR study of a heterogeneous biocatalyst using fast MAS at high magnetic fields
AU - Varghese, Sabu
AU - Halling, Peter J.
AU - Häussinger, Daniel
AU - Wimperis, Stephen
PY - 2018/3/14
Y1 - 2018/3/14
N2 - Nuclear magnetic resonance (NMR) is a powerful tool for investigating atomic-scale structure in heterogeneous or composite materials where long-range order is absent. In this work solid-state 1H and 1H-detected NMR experiments were performed with fast magic angle spinning (νR = 75 kHz) and at high magnetic fields (B0 = 20 T) and used to gain structural insight into a heterogeneous biocatalyst consisting of an enzyme, human carbonic anhydrase II (hCA II), covalently immobilized on epoxy-functionalized silica. Two-dimensional 1H-1H NOESY-type correlation experiments were able to provide information on 1H environments in silica, epoxy-silica and the immobilized enzyme. Two distinct signals originating from water protons were observed: water associated with the surface of the silica and the water associated with the immobilized enzyme. Additional two-dimensional 1H-1H double–single quantum (DQ-SQ) correlation experiments suggested that the immobilized enzyme is not in close contact with the silica surface. Most significantly, comparison of two-dimensional 1H-15N spectra of the immobilized enzyme and the solution-state enzyme confirmed that the structural integrity of the protein is well preserved upon covalent immobilization.
AB - Nuclear magnetic resonance (NMR) is a powerful tool for investigating atomic-scale structure in heterogeneous or composite materials where long-range order is absent. In this work solid-state 1H and 1H-detected NMR experiments were performed with fast magic angle spinning (νR = 75 kHz) and at high magnetic fields (B0 = 20 T) and used to gain structural insight into a heterogeneous biocatalyst consisting of an enzyme, human carbonic anhydrase II (hCA II), covalently immobilized on epoxy-functionalized silica. Two-dimensional 1H-1H NOESY-type correlation experiments were able to provide information on 1H environments in silica, epoxy-silica and the immobilized enzyme. Two distinct signals originating from water protons were observed: water associated with the surface of the silica and the water associated with the immobilized enzyme. Additional two-dimensional 1H-1H double–single quantum (DQ-SQ) correlation experiments suggested that the immobilized enzyme is not in close contact with the silica surface. Most significantly, comparison of two-dimensional 1H-15N spectra of the immobilized enzyme and the solution-state enzyme confirmed that the structural integrity of the protein is well preserved upon covalent immobilization.
KW - solid-state NMR
KW - biocatalysis
KW - covalent immobilization
KW - epoxy-functionalized silica
KW - hCA II
KW - heterogeneous biocatalysts
KW - human carbonic anhydrase II
KW - immobilized enzymes
UR - http://www.scopus.com/inward/record.url?scp=85044458032&partnerID=8YFLogxK
U2 - 10.1016/j.ssnmr.2018.03.003
DO - 10.1016/j.ssnmr.2018.03.003
M3 - Article
VL - 92
SP - 7
EP - 11
ER -