Two-dimensional 1H and 1H-detected NMR study of a heterogeneous biocatalyst using fast MAS at high magnetic fields

Sabu Varghese, Peter J. Halling, Daniel Häussinger, Stephen Wimperis

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

LanguageEnglish
Pages7-11
Number of pages5
JournalSolid State Nuclear Magnetic Resonance
Volume92
Early online date14 Mar 2018
DOIs
Publication statusE-pub ahead of print - 14 Mar 2018

Fingerprint

Biocatalysts
Immobilized Enzymes
Silicon Dioxide
enzymes
Enzymes
Nuclear magnetic resonance
Magnetic fields
Silica
nuclear magnetic resonance
silicon dioxide
magnetic fields
Water
Carbonic Anhydrase II
Magic angle spinning
Carbonic anhydrase
carbonic anhydrase
water
Experiments
Structural integrity
Protons

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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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.",
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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 journalArticle

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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

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