Biosilica-Entrapped Enzymes Studied by Using Dynamic Nuclear-Polarization-Enhanced High-Field NMR Spectroscopy.
Enzymes are used as environmentally friendly catalysts in many industrial applications, and are frequently immobilized in a matrix to improve their chemical stability for long-term storage and reusability. Recently, it was shown that an atomic-level description of proteins immobilized in a biosilica matrix can be attained by examining their magic-angle spinning (MAS) NMR spectra. However, even though MAS NMR is an excellent tool for determining structure, it is severely hampered by sensitivity. In this work we provide the proof of principle that NMR characterization of biosilica-entrapped enzymes could be assisted by high-field dynamic nuclear polarization (DNP).
Magnetic Resonance Center (CERM) and Department of Chemistry "Ugo Schiff", University of Florence, 50019 Sesto Fiorentino (FI) (Italy).
Enrico
E
Ravera
Francis Bitter Magnet Laboratory and Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139 (USA).
Vladimir K
VK
Michaelis
Francis Bitter Magnet Laboratory and Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139 (USA).|Current address: Department of Chemistry, Laboratory of Inorganic Chemistry, ETH- Zürich, CH-8093 Zürich (Switzerland).
Ta-Chung
TC
Ong
Francis Bitter Magnet Laboratory and Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139 (USA).
Eric G
EG
Keeler
Magnetic Resonance Center (CERM) and Department of Chemistry "Ugo Schiff", University of Florence, 50019 Sesto Fiorentino (FI) (Italy).|Current address: Giotto Biotech Srl, Via Madonna del Piano 6, 50019 Sesto Fiorentino (FI) (Italy).
Tommaso
T
Martelli
Magnetic Resonance Center (CERM) and Department of Chemistry "Ugo Schiff", University of Florence, 50019 Sesto Fiorentino (FI) (Italy).
Marco
M
Fragai
Francis Bitter Magnet Laboratory and Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, 02139 (USA).
Robert G
RG
Griffin
Magnetic Resonance Center (CERM) and Department of Chemistry "Ugo Schiff", University of Florence, 50019 Sesto Fiorentino (FI) (Italy).
Claudio
C
Luchinat
DNP; biomaterials; biosilica; enzyme immobilization; solid-state NMR