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

Varghese, Sabu and Halling, Peter J. and Häussinger, Daniel and Wimperis, Stephen (2018) Two-dimensional 1H and 1H-detected NMR study of a heterogeneous biocatalyst using fast MAS at high magnetic fields. Solid State Nuclear Magnetic Resonance, 92. pp. 7-11. ISSN 0926-2040

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Abstract

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.

Item Type:
Journal Article
Journal or Publication Title:
Solid State Nuclear Magnetic Resonance
Additional Information:
This is the author’s version of a work that was accepted for publication in Solid State Nuclear Magnetic Resonance. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Solid State Nuclear Magnetic Resonance, 92, 2018 DOI: 10.1016/j.ssnmr.2018.03.003
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/3100/3108
Subjects:
ID Code:
124082
Deposited By:
Deposited On:
16 Mar 2018 11:54
Refereed?:
Yes
Published?:
Published
Last Modified:
27 Sep 2020 04:25